In yet another aspect, the present disclosure provides a polynucleotide encoding an AAV capsid variant, wherein the AAV capsid variant comprises: (i) the amino acid sequence of SPHSKA (SEQ ID NO: 941); (ii) an amino acid sequence comprising at least one, two, or three, but no more than four different amino acids, relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941); (iii) an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941); or (iv) at least 3, 4, 5, 6, 7, 8, or 9 consecutive amino acids from the amino acid sequence of SPHSKA (SEQ ID NO: 941). In some embodiments, the amino acid sequence of (i), (ii), (iii), and/or (iv) is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.

In another aspect, the present disclosure provides a polynucleotide encoding an AAV capsid variant, wherein the AAV capsid variant comprises: (i) the amino acid sequence of HDSPHK (SEQ ID NO: 2); (ii) an amino acid sequence comprising at least one, two, or three, but no more than four different amino acids, relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2); (iii) an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2); or (iv) at least 3, 4, 5, or 6 consecutive amino acids from the amino acid sequence of HDSPHK (SEQ ID NO: 2). In some embodiments, the amino acid sequence of (i), (ii), (iii), and/or (iv) is present immediately subsequent to position 453, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.

In yet another aspect, the present disclosure provides an AAV capsid variant comprising the amino acid sequence of SPH, wherein the amino acid sequence is present immediately subsequent to position 455, numbered according to the amino acid sequence of any one of SEQ ID NOs: 36-59, 138, 981, or 982.

In yet another aspect, the present disclosure provides an AAV capsid variant comprising the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to position 455, numbered according to the amino acid sequence of SEQ ID NO: 138.

In yet another aspect, the present disclosure provides an AAV capsid variant comprising the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to position 455, numbered according to the amino acid sequence of SEQ ID NO: 981.

In yet another aspect, the present disclosure provides an AAV capsid variant comprising the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein the amino acid sequence is present immediately subsequent to position 453, numbered according to the amino acid sequence of SEQ ID NO: 138.

In yet another aspect, the present disclosure provides an AAV capsid variant comprising the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein the amino acid sequence is present immediately subsequent to position 453, numbered according to the amino acid sequence of SEQ ID NO: 982.

In yet another aspect, the present disclosure provides an AAV capsid variant comprising the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present in loop IV, relative to a reference sequence of SEQ ID NO: 138. In some embodiments, loop IV comprises positions 449-460, numbered according to SEQ ID NO: 138.

In yet another aspect, the present disclosure provides an AAV capsid variant comprising the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein the amino acid sequence is present in loop IV, relative to a reference sequence of SEQ ID NO: 138. In some embodiments, loop IV comprises positions 449-460, numbered according to SEQ ID NO: 138.

In yet another aspect, the present disclosure provides an AAV particle comprising an AAV capsid variant, described herein. In some embodiments, the AAV particle comprises a nucleic acid sequence encoding a payload. In some embodiments, the AAV particle further comprises a viral genome comprising a promoter operably linked to the nucleic acid encoding the payload.

In yet another aspect, the present disclosure provides a method of making an AAV particle comprising an AAV capsid variant described herein. The method comprises providing a host cell comprising a viral genome and incubating the host cell under conditions suitable to enclose the viral genome in the AAV capsid variant, e.g., an AAV capsid variant described herein, thereby making the AAV particle.

In yet another aspect, the present disclosure provides a method of delivering a payload to a cell or tissue (e.g., a CNS cell, a CNS tissue, a liver cell, or a liver tissue). The method comprising administering an effective amount of an AAV particle comprising an AAV capsid variant described herein.

In yet another aspect, the present disclosure provides a method of treating a subject having or diagnosed with having a genetic disorder, e.g., a monogenic disorder or a polygenic disorder. The method comprising administering to the subject an effective amount an AAV particle comprising an AAV capsid variant described herein.

In yet another aspect, the present disclosure provides a method of treating a subject having or diagnosed with having neurological, e.g., a neurodegenerative, disorder. The method comprising administering an effective amount of an AAV particle comprising an AAV capsid variant described herein.

In yet another aspect, the present disclosure provides a method of treating a subject having or diagnosed with having a neuro-oncological disorder. The method comprising administering an effective amount of an AAV particle comprising an AAV capsid variant described herein.

Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following enumerated embodiments.

ENUMERATED EMBODIMENTS

1. An AAV capsid variant, comprising an amino acid sequence having the following formula: [N1]-[N2]-[N3], wherein:

- (i) optionally [N1] comprises X1, X2, and X3, wherein at least one of X1, X2, or X3 is G;
- (ii) [N2] comprises the amino acid sequence of SPH;
- (ii) [N3] comprises X4, X5, and X6, wherein at least one of X4, X5, or X6 is a basic amino acid, e.g., a K or R.
  
  2. The AAV capsid variant of embodiment 1, wherein X4, X5, or both of [N3] is a K.
  
  3. The AAV capsid variant of embodiment 1 or 2, wherein X4, X5, or X6 of [N3] is an R.
  
  4. The AAV capsid variant of any one of embodiments 1-3, wherein:
- (a) position X4 of [N3] is: K, S, A, V, T, G, F, W, V, N, or R;
- (b) position X5 of [N3] is: S, K, T, F, I, L, Y, H, M, or R; and/or
- (c) position X6 of [N3] is: G, A, R, M, I, N, T, Y, D, P, V, L, E, W, N, Q, K, or S;
- optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (a)-(c).
  
  5. The AAV capsid variant of any one of embodiments 1-4, wherein [N3] comprises SK, KA, KS, AR, RM, VK, AS, SR, VK, KR, KK, KN, VR, RS, RK, KT, TS, KF, FG, KI, IG, KL, LG, TT, TY, KY, YG, KD, KP, TR, RG, VR, GA, SL, SS, FL, WK, SA, RA, LR, KW, RR, GK, TK, NK, AK, KV, KG, KH, KM, TG, SE, SV, SW, SN, HG, SQ, LW, MG, MA, or SG.
  
  6. The AAV capsid variant of any one of embodiments 1-5, wherein [N3] is or comprises SKA, KSG, ARM, VKS, ASR, VKI, KKN, VRM, RKA, KTS, KFG, KIG, KLG, KTT, KTY, KYG, SKD, SKP, TRG, VRG, KRG, GAR, KSA, KSR, SKL, SRA, SKR, SLR, SRG, SSR, FLR, SKW, SKS, WKA, VRR, SKV, SKT, SKG, GKA, TKA, NKA, SKL, SKN, AKA, KTG, KSL, KSE, KSV, KSW, KSN, KHG, KSQ, KSK, KLW, WKG, KMG, KMA, or RSG.
  
  7. The AAV capsid variant of any one of embodiments 1-6, wherein [N2]-[N3] comprises SPHSK (SEQ ID NO: 4701), SPHKS (SEQ ID NO: 4704), SPHAR (SEQ ID NO: 4705), SPHVK (SEQ ID NO: 4706), SPHAS (SEQ ID NO: 4707), SPHKK (SEQ ID NO: 4708), SPHVR (SEQ ID NO: 4709), SPHRK (SEQ ID NO: 4710), SPHKT (SEQ ID NO: 4711), SPHKF (SEQ ID NO: 4712), SPHKI (SEQ ID NO: 4713), SPHKL (SEQ ID NO: 4714), SPHKY (SEQ ID NO: 4715), SPHTR (SEQ ID NO: 4716), SPHKR (SEQ ID NO: 4717), SPHGA (SEQ ID NO: 4718), SPHSR (SEQ ID NO: 4719), SPHSL (SEQ ID NO: 4720), SPHSS (SEQ ID NO: 4721), SPHFL (SEQ ID NO: 4722), SPHWK (SEQ ID NO: 4723), SPHGK (SEQ ID NO: 4724), SPHTK (SEQ ID NO: 4725), SPHNK (SEQ ID NO: 4726), SPHAK (SEQ ID NO: 4727), SPHKH (SEQ ID NO: 4728), SPHKM (SEQ ID NO: 4729), or SPHRS (SEQ ID NO: 4730).
  
  8. The AAV capsid variant of any one of embodiments 1-7, wherein [N2]-[N3] is or comprises:

(i)

(SEQ ID NO: 941)

SPHSKA,

(SEQ ID NO: 946)

SPHKSG,

(SEQ ID NO: 947)

SPHARM,

(SEQ ID NO: 948)

SPHVKS,

(SEQ ID NO: 949)

SPHASR,

(SEQ ID NO: 950)

SPHVKI,

(SEQ ID NO: 954)

SPHKKN

(SEQ ID NO: 955)

SPHVRM,

(SEQ ID NO: 956)

SPHRKA,

(SEQ ID NO: 957)

SPHKFG,

(SEQ ID NO: 958)

SPHKIG,

(SEQ ID NO: 959)

SPHKLG,

(SEQ ID NO: 963)

SPHKTS,

(SEQ ID NO: 964)

SPHKTT,

(SEQ ID NO: 965)

SPHKTY,

(SEQ ID NO: 966)

SPHKYG,

(SEQ ID NO: 967)

SPHSKD,

(SEQ ID NO: 968)

SPHSKP,

(SEQ ID NO: 972)

SPHTRG,

(SEQ ID NO: 973)

SPHVRG,

(SEQ ID NO: 974)

SPHKRG,

(SEQ ID NO: 975)

SPHGAR,

(SEQ ID NO: 977)

SPHKSA,

(SEQ ID NO: 951)

SPHKSR,

(SEQ ID NO: 960)

SPHSKL,

(SEQ ID NO: 969)

SPHSRA,

(SEQ ID NO: 978)

SPHSKR,

(SEQ ID NO: 952)

SPHSLR,

(SEQ ID NO: 961)

SPHSRG,

(SEQ ID NO: 970)

SPHSSR,

(SEQ ID NO: 979)

SPHFLR,

(SEQ ID NO: 953)

SPHSKW,

(SEQ ID NO: 962)

SPHSKS,

(SEQ ID NO: 971)

SPHWKA,

(SEQ ID NO: 980)

SPHVRR,

(SEQ ID NO: 4731)

SPHSKT,

(SEQ ID NO: 4732)

SPHSKG,

(SEQ ID NO: 4733)

SPHGKA,

(SEQ ID NO: 4734)

SPHNKA,

(SEQ ID NO: 4735)

SPHSKN,

(SEQ ID NO: 4736)

SPHAKA,

(SEQ ID NO: 4737)

SPHSKV,

(SEQ ID NO: 4738)

SPHKTG,

(SEQ ID NO: 4739)

SPHTKA,

(SEQ ID NO: 4740)

SPHKSL,

(SEQ ID NO: 4741)

SPHKSE,

(SEQ ID NO: 4742)

SPHKSV,

(SEQ ID NO: 4743)

SPHKSW,

(SEQ ID NO: 4744)

SPHKSN,

(SEQ ID NO: 4745)

SPHKHG,

(SEQ ID NO: 4746)

SPHKSQ,

(SEQ ID NO: 4747)

SPHKSK,

(SEQ ID NO: 4748)

SPHKLW,

(SEQ ID NO: 4749)

SPHWKG,

(SEQ ID NO: 4750)

SPHKMG,

(SEQ ID NO: 4751)

SPHKMA,

or

(SEQ ID NO: 976)

SPHRSG;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, or 5 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  9. The AAV capsid variant of any one of embodiments 1-8, which comprises an amino acid other than G at position 453 (e.g., V, R, D, E, M, T, I, S, A, N, L, K, H, P, W, or C), an amino acid other than S at position 454 (V, L, N, D, H, R, P, G, T, I, A, E, Y, M, or Q), and/or a G at position 455 (e.g., C, L, D, E, Y, H, V, A, N, P, or S), numbered according to any one of SEQ ID NOs: 36-59, 138, 981, 982.
  
  10. The AAV capsid variant of any one of embodiments 1-8, which comprises the amino acid G at position 453, the amino acid S at position 454, and the amino acid G at position 455, numbered according to SEQ ID NO: 138 or 981.
  
  11. The AAV capsid variant of any one of embodiments 1-9, which comprises the amino acid G at position 453, the amino acid H at position 454, and the amino acid D at position 455, numbered according to SEQ ID NO: 138 or 982.
  
  12. The AAV capsid variant of any one of embodiments 1-11, wherein [N1] comprises X1, X2, and X3, wherein at least one of X1, X2, or X3 is G.
  
  13. The AAV capsid variant of any one of embodiments 1-12, wherein:
- (a) position X1 of [N1] is: G, V, R, D, E, M, T, I, S, A, N, L, K, H, P, W, or C;
- (b) position X2 of [N1] is: S, V, L, N, D, H, R, P, G, T, I, A, E, Y, M, or Q; and/or
- (c) position X3 of [N1] is: G, C, L, D, E, Y, H, V, A, N, P, or S;
- optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (a)-(c).
  
  14. The AAV capsid variant of any one of embodiments 1-13, wherein [N1] comprises GS, SG, GH, HD, GQ, QD, VS, CS, GR, RG, QS, SH, MS, RN, TS, IS, GP, ES, SS, GN, AS, NS, LS, GG, KS, GT, PS, RS, GI, WS, DS, ID, GL, DA, DG, ME, EN, KN, KE, AI, NG, PG, TG, SV, IG, LG, AG, EG, SA, YD, HE, HG, RD, ND, PD, MG, QV, DD, HN, HP, GY, GM, GD, or HS.
  
  15. The AAV capsid variant of any one of embodiments 1-14, wherein [N1] is or comprises GSG, GHD, GQD, VSG, CSG, GRG, CSH, GQS, GSH, RVG, GSC, GLL, GDD, GHE, GNY, MSG, RNG, TSG, ISG, GPG, ESG, SSG, GNG, ASG, NSG, LSG, GGG, KSG, HSG, GTG, PSG, GSV, RSG, GIG, WSG, DSG, IDG, GLG, DAG, DGG, MEG, ENG, GSA, KNG, KEG, AIG, GYD, GHG, GRD, GND, GPD, GMG, GQV, GHN, GHP, or GHS.
  
  16. The AAV capsid variant of any one of embodiments 1-15, wherein [N1]-[N2] comprises:

(i)

(SEQ ID NO: 4752)

SGSPH,

(SEQ ID NO: 4703)

HDSPH,

(SEQ ID NO: 4753)

QDSPH,

(SEQ ID NO: 4754)

RGSPH,

(SEQ ID NO: 4755)

SHSPH,

(SEQ ID NO: 4756)

QSSPH,

(SEQ ID NO: 4757)

DDSPH,

(SEQ ID NO: 4758)

HESPH,

(SEQ ID NO: 4759)

NYSPH,

(SEQ ID NO: 4760)

VGSPH,

(SEQ ID NO: 4761)

SCSPH,

(SEQ ID NO: 4762)

LLSPH,

(SEQ ID NO: 4763)

NGSPH,

(SEQ ID NO: 4764)

PGSPH,

(SEQ ID NO: 4765)

GGSPH,

(SEQ ID NO: 4766)

TGSPH,

(SEQ ID NO: 4767)

SVSPH

(SEQ ID NO: 4768)

IGSPH,

(SEQ ID NO: 4769)

DGSPH,

(SEQ ID NO: 4770)

LGSPH,

(SEQ ID NO: 4771)

AGSPH,

(SEQ ID NO: 4772)

EGSPH,

(SEQ ID NO: 4773)

SASPH,

(SEQ ID NO: 4774)

YDSPH,

(SEQ ID NO: 4775)

HGSPH,

(SEQ ID NO: 4776)

RDSPH,

(SEQ ID NO: 4777)

NDSPH

(SEQ ID NO: 4778)

PDSPH,

(SEQ ID NO: 4779)

MGSPH,

(SEQ ID NO: 4780)

QVSPH,

(SEQ ID NO: 4781)

HNSPH,

(SEQ ID NO: 4782)

HPSPH,

or

(SEQ ID NO: 4783)

HSSPH;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, or 4 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  17. The AAV capsid variant of any one of embodiments 1-16, wherein [N1]-[N2] is or comprises:

(i)

(SEQ ID NO: 4695)

GSGSPH,

(SEQ ID NO: 4784)

GHDSPH,

(SEQ ID NO: 4785)

GQDSPH,

(SEQ ID NO: 4786)

VSGSPH,

(SEQ ID NO: 4787)

CSGSPH,

(SEQ ID NO: 4788)

GRGSPH,

(SEQ ID NO: 4789)

CSHSPH,

(SEQ ID NO: 4790)

GQSSPH,

(SEQ ID NO: 4791)

GSHSPH,

(SEQ ID NO: 4792)

GDDSPH,

(SEQ ID NO: 4793)

GHESPH,

(SEQ ID NO: 4794)

GNYSPH,

(SEQ ID NO: 4795)

RVGSPH,

(SEQ ID NO: 4796)

GSCSPH,

(SEQ ID NO: 4797)

GLLSPH,

(SEQ ID NO: 4798)

MSGSPH,

(SEQ ID NO: 4799)

RNGSPH,

(SEQ ID NO: 4800)

TSGSPH,

(SEQ ID NO: 4801)

ISGSPH,

(SEQ ID NO: 4802)

GPGSPH,

(SEQ ID NO: 4803)

ESGSPH,

(SEQ ID NO: 4804)

SSGSPH,

(SEQ ID NO: 4805)

GNGSPH,

(SEQ ID NO: 4806)

ASGSPH,

(SEQ ID NO: 4807)

NSGSPH,

(SEQ ID NO: 4808)

LSGSPH,

(SEQ ID NO: 4809)

GGGSPH,

(SEQ ID NO: 4810)

KSGSPH,

(SEQ ID NO: 4811)

HSGSPH,

(SEQ ID NO: 4812)

GTGSPH,

(SEQ ID NO: 4813)

PSGSPH,

(SEQ ID NO: 4814)

GSVSPH,

(SEQ ID NO: 4815)

RSGSPH,

(SEQ ID NO: 4816)

GIGSPH,

(SEQ ID NO: 4817)

WSGSPH,

(SEQ ID NO: 4818)

DSGSPH,

(SEQ ID NO: 4819)

IDGSPH,

(SEQ ID NO: 4820)

GLGSPH,

(SEQ ID NO: 4821)

DAGSPH,

(SEQ ID NO: 4822)

DGGSPH,

(SEQ ID NO: 4823)

MEGSPH,

(SEQ ID NO: 4824)

ENGSPH,

(SEQ ID NO: 4825)

GSASPH,

(SEQ ID NO: 4826)

KNGSPH,

(SEQ ID NO: 4827)

KEGSPH,

(SEQ ID NO: 4828)

AIGSPH,

(SEQ ID NO: 4829)

GYDSPH,

(SEQ ID NO: 4830)

GHGSPH,

(SEQ ID NO: 4831)

GRDSPH,

(SEQ ID NO: 4832)

GNDSPH,

(SEQ ID NO: 4833)

GPDSPH,

(SEQ ID NO: 4834)

GMGSPH,

(SEQ ID NO: 4835)

GQVSPH,

(SEQ ID NO: 4836)

GHNSPH,

(SEQ ID NO: 4837)

GHPSPH,

or

(SEQ ID NO: 4838)

GHSSPH;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, or 5 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  18. The AAV capsid variant of any one of embodiments 1-17, wherein [N1]-[N2]-[N3] comprises:

(i)

(SEQ ID NO: 4839)

SGSPHSK,

(SEQ ID NO: 4840)

HDSPHKS,

(SEQ ID NO: 4841)

SGSPHAR,

(SEQ ID NO: 4842)

SGSPHVK,

(SEQ ID NO: 4843)

QDSPHKS,

(SEQ ID NO: 4844)

SGSPHKK,

(SEQ ID NO: 4845)

SGSPHVR,

(SEQ ID NO: 4846)

SGSPHAS,

(SEQ ID NO: 4847)

SGSPHRK,

(SEQ ID NO: 4848)

SGSPHKT,

(SEQ ID NO: 4849)

SHSPHKS,

(SEQ ID NO: 4850)

QSSPHRS,

(SEQ ID NO: 4851)

RGSPHAS,

(SEQ ID NO: 4852)

RGSPHSK,

(SEQ ID NO: 4853)

SGSPHKF,

(SEQ ID NO: 4854)

SGSPHKI,

(SEQ ID NO: 4855)

SGSPHKL,

(SEQ ID NO: 4856)

SGSPHKY,

(SEQ ID NO: 4857)

SGSPHTR,

(SEQ ID NO: 4858)

SHSPHKR,

(SEQ ID NO: 4859)

SGSPHGA,

(SEQ ID NO: 4860)

HDSPHKR,

(SEQ ID NO: 4861)

DDSPHKS,

(SEQ ID NO: 4862)

HESPHKS,

(SEQ ID NO: 4863)

NYSPHKI,

(SEQ ID NO: 4864)

SGSPHSR,

(SEQ ID NO: 4865)

SGSPHSL,

(SEQ ID NO: 4866)

SGSPHSS,

(SEQ ID NO: 4867)

VGSPHSK,

(SEQ ID NO: 4868)

SCSPHRK,

(SEQ ID NO: 4869)

SGSPHFL,

(SEQ ID NO: 4870)

LLSPHWK,

(SEQ ID NO: 4871)

NGSPHSK,

(SEQ ID NO: 4872)

PGSPHSK,

(SEQ ID NO: 4873)

GGSPHSK,

(SEQ ID NO: 4874)

TGSPHSK,

(SEQ ID NO: 4875)

SVSPHGK,

(SEQ ID NO: 4876)

SGSPHTK,

(SEQ ID NO: 4877)

IGSPHSK,

(SEQ ID NO: 4878)

DGSPHSK,

(SEQ ID NO: 4879)

SGSPHNK,

(SEQ ID NO: 4880)

LGSPHSK,

(SEQ ID NO: 4881)

AGSPHSK,

(SEQ ID NO: 4882)

EGSPHSK,

(SEQ ID NO: 4883)

SASPHSK,

(SEQ ID NO: 4884)

SGSPHAK,

(SEQ ID NO: 4885)

HDSPHKI,

(SEQ ID NO: 4886)

YDSPHKS,

(SEQ ID NO: 4887)

HDSPHKT,

(SEQ ID NO: 4888)

RGSPHKR,

(SEQ ID NO: 4889)

HGSPHSK,

(SEQ ID NO: 4890)

RDSPHKS,

(SEQ ID NO: 4891)

NDSPHKS,

(SEQ ID NO: 4892)

QDSPHKI,

(SEQ ID NO: 4893)

PDSPHKI,

(SEQ ID NO: 4894)

PDSPHKS,

(SEQ ID NO: 4895)

MGSPHSK,

(SEQ ID NO: 4896)

HDSPHKH,

(SEQ ID NO: 4897)

QVSPHKS,

(SEQ ID NO: 4898)

HNSPHKS,

(SEQ ID NO: 4899)

NGSPHKR,

(SEQ ID NO: 4900)

HDSPHKY,

(SEQ ID NO: 4901)

NDSPHKI,

(SEQ ID NO: 4902)

HDSPHKL,

(SEQ ID NO: 4903)

HPSPHWK,

(SEQ ID NO: 4904)

HDSPHKM,

or

(SEQ ID NO: 4905)

HSSPHRS;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, or 6 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  19. The AAV capsid variant of any one of embodiments 1-18, wherein [N1]-[N2]-[N3] is or comprises:

(i)

(SEQ ID NO: 4697)

GSGSPHSKA,

(SEQ ID NO: 4698)

GHDSPHKSG,

(SEQ ID NO: 4906)

GSGSPHARM

(SEQ ID NO: 4907)

GSGSPHVKS,

(SEQ ID NO: 4908)

GQDSPHKSG,

(SEQ ID NO: 4909)

GSGSPHASR,

(SEQ ID NO: 4910)

GSGSPHVKI,

(SEQ ID NO: 4911)

GSGSPHKKN,

(SEQ ID NO: 4912)

GSGSPHVRM,

(SEQ ID NO: 4913)

VSGSPHSKA,

(SEQ ID NO: 4914)

CSGSPHSKA,

(SEQ ID NO: 4915)

GSGSPHRKA,

(SEQ ID NO: 4916)

CSGSPHKTS,

(SEQ ID NO: 4917)

CSHSPHKSG,

(SEQ ID NO: 4918)

GQSSPHRSG,

(SEQ ID NO: 4919)

GRGSPHASR,

(SEQ ID NO: 4920)

GRGSPHSKA,

(SEQ ID NO: 4921)

GSGSPHKFG,

(SEQ ID NO: 4922)

GSGSPHKIG,

(SEQ ID NO: 4923)

GSGSPHKLG,

(SEQ ID NO: 4924)

GSGSPHKTS,

(SEQ ID NO: 4925)

GSGSPHKTT,

(SEQ ID NO: 4926)

GSGSPHKTY,

(SEQ ID NO: 4927)

GSGSPHKYG,

(SEQ ID NO: 4928)

GSGSPHSKD,

(SEQ ID NO: 4929)

GSGSPHSKP,

(SEQ ID NO: 4930)

GSGSPHTRG,

(SEQ ID NO: 4931)

GSGSPHVRG,

(SEQ ID NO: 4932)

GSHSPHKRG,

(SEQ ID NO: 4933)

GSHSPHKSG,

(SEQ ID NO: 4934)

VSGSPHASR,

(SEQ ID NO: 4935)

VSGSPHGAR,

(SEQ ID NO: 4936)

VSGSPHKFG,

(SEQ ID NO: 4937)

GHDSPHKRG,

(SEQ ID NO: 4938)

GDDSPHKSG,

(SEQ ID NO: 4939)

GHESPHKSA,

(SEQ ID NO: 4940)

GHDSPHKSA,

(SEQ ID NO: 4941)

GNYSPHKIG,

(SEQ ID NO: 4942)

GHDSPHKSR,

(SEQ ID NO: 4943)

GSGSPHSKL,

(SEQ ID NO: 4944)

GSGSPHSRA,

(SEQ ID NO: 4945)

GSGSPHSKR,

(SEQ ID NO: 4946)

GSGSPHSLR,

(SEQ ID NO: 4947)

GSGSPHSRG,

(SEQ ID NO: 4948)

GSGSPHSSR,

(SEQ ID NO: 4949)

RVGSPHSKA,

(SEQ ID NO: 4950)

GSCSPHRKA,

(SEQ ID NO: 4951)

GSGSPHFLR,

(SEQ ID NO: 4952)

GSGSPHSKW,

(SEQ ID NO: 4953)

GSGSPHSKS,

(SEQ ID NO: 4954)

GLLSPHWKA,

(SEQ ID NO: 4955)

GSGSPHVRR,

(SEQ ID NO: 4956)

GSGSPHSKV,

(SEQ ID NO: 4957)

MSGSPHSKA,

(SEQ ID NO: 4958)

RNGSPHSKA,

(SEQ ID NO: 4959)

TSGSPHSKA,

(SEQ ID NO: 4960)

ISGSPHSKA,

(SEQ ID NO: 4961)

GPGSPHSKA,

(SEQ ID NO: 4962)

GSGSPHSKT

(SEQ ID NO: 4963)

ESGSPHSKA,

(SEQ ID NO: 4964)

SSGSPHSKA,

(SEQ ID NO: 4965)

GNGSPHSKA,

(SEQ ID NO: 4966)

ASGSPHSKA,

(SEQ ID NO: 4967)

NSGSPHSKA,

(SEQ ID NO: 4968)

LSGSPHSKA,

(SEQ ID NO: 4969)

GGGSPHSKA,

(SEQ ID NO: 4970)

KSGSPHSKA

(SEQ ID NO: 4971)

GGGSPHSKS,

(SEQ ID NO: 4972)

GSGSPHSKG,

(SEQ ID NO: 4973)

HSGSPHSKA,

(SEQ ID NO: 4974)

GTGSPHSKA,

(SEQ ID NO: 4975)

PSGSPHSKA,

(SEQ ID NO: 4976)

GSVSPHGKA,

(SEQ ID NO: 4977)

RSGSPHSKA,

(SEQ ID NO: 4978)

GSGSPHTKA,

(SEQ ID NO: 4979)

GIGSPHSKA,

(SEQ ID NO: 4980)

WSGSPHSKA,

(SEQ ID NO: 4981)

DSGSPHSKA,

(SEQ ID NO: 4982)

IDGSPHSKA,

(SEQ ID NO: 4983)

GSGSPHNKA,

(SEQ ID NO: 4984)

GLGSPHSKS,

(SEQ ID NO: 4985)

DAGSPHSKA,

(SEQ ID NO: 4986)

DGGSPHSKA,

(SEQ ID NO: 4987)

MEGSPHSKA,

(SEQ ID NO: 4988)

ENGSPHSKA,

(SEQ ID NO: 4989)

GSASPHSKA,

(SEQ ID NO: 4990)

GNGSPHSKS,

(SEQ ID NO: 4991)

KNGSPHSKA,

(SEQ ID NO: 4992)

KEGSPHSKA,

(SEQ ID NO: 4993)

AIGSPHSKA,

(SEQ ID NO: 4994)

GSGSPHSKN

(SEQ ID NO: 4995)

GSGSPHAKA,

(SEQ ID NO: 4996)

GHDSPHKIG,

(SEQ ID NO: 4997)

GYDSPHKSG,

(SEQ ID NO: 4998)

GHESPHKSG,

(SEQ ID NO: 4999)

GHDSPHKTG,

(SEQ ID NO: 5000)

GRGSPHKRG,

(SEQ ID NO: 4908)

GQDSPHKSG,

(SEQ ID NO: 5001)

GHDSPHKSL

(SEQ ID NO: 5002)

GHGSPHSKA,

(SEQ ID NO: 5003)

GHDSPHKSE,

(SEQ ID NO: 4913)

VSGSPHSKA,

(SEQ ID NO: 5004)

GRDSPHKSG,

(SEQ ID NO: 5005)

GNDSPHKSV,

(SEQ ID NO: 5006)

GQDSPHKIG,

(SEQ ID NO: 5007)

GHDSPHKSV,

(SEQ ID NO: 5008)

GPDSPHKIG,

(SEQ ID NO: 5009)

GPDSPHKSG,

(SEQ ID NO: 5010)

GHDSPHKSW,

(SEQ ID NO: 5011)

GHDSPHKSN,

(SEQ ID NO: 5012)

GMGSPHSKT,

(SEQ ID NO: 5013)

GHDSPHKHG,

(SEQ ID NO: 5014)

GQVSPHKSG,

(SEQ ID NO: 5015)

GDDSPHKSV,

(SEQ ID NO: 5016)

GHNSPHKSG,

(SEQ ID NO: 5017)

GNGSPHKRG,

(SEQ ID NO: 5018)

GHDSPHKYG,

(SEQ ID NO: 5019)

GHDSPHKSQ,

(SEQ ID NO: 5020)

GNDSPHKIG,

(SEQ ID NO: 5021)

GHDSPHKSK,

(SEQ ID NO: 5022)

GHDSPHKLW,

(SEQ ID NO: 5023)

GHPSPHWKG,

(SEQ ID NO: 5024)

GHDSPHKMG,

(SEQ ID NO: 5025)

GHDSPHKMA,

or

(SEQ ID NO: 5026)

GHSSPHRSG;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, 6, 7, or 8 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  20. The AAV capsid variant of any one of embodiments 1-19, wherein [N3] comprises SK, KA, KS, or SG.
  
  21. The AAV capsid variant of any one of embodiments 1-20, wherein [N3] is or comprises SKA, KSG, or KYG.
  
  22. The AAV capsid variant of any one of embodiments 1-21, wherein [N2]-[N3] comprises SPHSK (SEQ ID NO: 4701), SPHKS (SEQ ID NO: 4704), or SPHKY (SEQ ID NO: 4715).
  
  23. The AAV capsid variant of any one of embodiments 1-22, wherein [N2]-[N3] is or comprises SPHSKA (SEQ ID NO: 941).
  
  24. The AAV capsid variant of any one of embodiments 1-22, wherein [N2]-[N3] is or comprises SPHKSG (SEQ ID NO: 946).
  
  25. The AAV capsid variant of any one of embodiments 1-22, wherein [N2]-[N3] is or comprises SPHKYG (SEQ ID NO: 966).
  
  26. The AAV capsid variant of any one of embodiments 1-25, wherein [N1] comprises GS, SG, GH, or HD.
  
  27. The AAV capsid variant of any one of embodiments 1-26, wherein [N1] is or comprises GSG.
  
  28. The AAV capsid variant of any one of embodiments 1-26, wherein [N1] is or comprises GHD.
  
  29. The AAV capsid variant of any one of embodiments 1-23 or 26-27, wherein [N1]-[N2]-[N3] comprises SGSPHSK (SEQ ID NO: 4839).
  
  30. The AAV capsid variant of any one of embodiments 1-22, 24, 26, or 28, wherein [N1]-[N2]-[N3] comprises HDSPHKS (SEQ ID NO: 4840).
  
  31. The AAV capsid variant of any one of embodiments 1-22 or 25-27, wherein [N1]-[N2]-[N3] comprises SGSPHKYG (SEQ ID NO: 5027).
  
  32. The AAV capsid variant of any one of embodiments 1-8, 10, 12-23, 26-27, or 29, wherein [N1]-[N2]-[N3] is or comprises GSGSPHSKA (SEQ ID NO: 4697).
  
  33. The AAV capsid variant of any one of embodiments 1-9, 11-22, 24, 26, 28, or 30, wherein [N1]-[N2]-[N3] is or comprises GHDSPHKSG (SEQ ID NO: 4698).
  
  34. The AAV capsid variant of any one of embodiments 1-8, 10, 12-22, 25-27, or 31, wherein [N1]-[N2]-[N3] is or comprises GSGSPHKYG (SEQ ID NO: 4927).
  
  35. The AAV capsid variant of any one of embodiments 1-34, wherein [N1]-[N2]-[N3] replaces positions 453-455, numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  36. The AAV capsid variant of any one of embodiments 1-35, which comprises an amino acid other than Q at position 456 (e.g., W, K, R, G, L, V, S, P, H, K, I, M, A, E, or F), an amino acid other than N at position 457 (e.g., Y, C, K, T, H, R, D, V, S, P, G, W, E, F, A, I, M, Q, or L), an amino acid other than Q at position 458 (e.g., G, K, H, R, T, L, D, A, P, I, F, V, M, W, Y, S, E, N, or Y), and/or an amino acid other than Q at position 459 (e.g., H, L, R, W, K, A, P, E, M, I, S, G, N, Y, C, V, T, D, or V), relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  37. The AAV capsid variant of any one of embodiments 1-36, which comprises an amino acid other than Q at position 462 (e.g., W, K, R, G, L, V, S, P, H, K, I, M, A, E, or F), an amino acid other than N at position 463 (e.g., Y, C, K, T, H, R, D, V, S, P, G, W, E, F, A, I, M, Q, or L), an amino acid other than Q at position 464 (e.g., G, K, H, R, T, L, D, A, P, I, F, V, M, W, Y, S, E, N, or Y), and/or an amino acid other than Q at position 465 (e.g., H, L, R, W, K, A, P, E, M, I, S, G, N, Y, C, V, T, D, or V), relative to a reference sequence numbered according to SEQ ID NO: 981, 982, 36, 37, 39, 40, 42-46, 48, 49, 50, 52, 53, 56, or 57.
  
  38. The AAV capsid variant of any one of embodiments 1-37, which comprises:
- (a) the amino acid Q at position 456, the amino acid N at position 457, the amino acid Q at position 458, and/or the amino acid Q at position 459, relative to a reference sequence numbered according to SEQ ID NO: 138; or
- (b) the amino acid Q at position 462, the amino acid N at position 463, the amino acid Q at position 464, and/or the amino acid Q at position 465, relative to a reference sequence numbered according to SEQ ID NO: 981, 982, 36, 37, 39, 40, 42-46, 48, 49, 50, 52, 53, 56, or 57.
  
  39. The AAV capsid variant of any one of embodiments 1-38, which further comprises [N4], wherein [N4] comprises X7 X8 X9 X10, and wherein:
- (a) position X7 is: Q, W, K, R, G, L, V, S, P, H, K, I, M, A, E, or F;
- (b) position X8 is: N, Y, C, K, T, H, R, D, V, S, P, G, W, E, F, A, I, M, Q, or L;
- (c) position X9 is: Q, G, K, H, R, T, L, D, A, P, I, F, V, M, W, Y, S, E, N, or Y; and
- (d) position X10 is: Q, H, L, R, W, K, A, P, E, M, I, S, G, N, Y, C, V, T, D, or V;
- optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (a)-(d).
  
  40. The AAV capsid variant of embodiment 39, wherein:
- (a) position X7 of [N4] is Q or R;
- (b) position X8 of [N4] is N or R;
- (c) position X9 of [N4] is Q or R; and
- (d) position X10 of [N4] is Q, L, or R.
  
  41. The AAV capsid variant of embodiment 39 or 40, wherein [N4] is or comprises:

(i)

(SEQ ID NO: 5028)

QNQQ,

(SEQ ID NO: 5029)

WNQQ,

(SEQ ID NO: 5030)

QYYV,

(SEQ ID NO: 5031)

RRQQ,

(SEQ ID NO: 5032)

GCGQ,

(SEQ ID NO: 5033)

LRQQ,

(SEQ ID NO: 5034)

RNQQ,

(SEQ ID NO: 5035)

VNQQ,

(SEQ ID NO: 5036)

FRLQ,

(SEQ ID NO: 5037)

FNQQ,

(SEQ ID NO: 5038)

LLQQ,

(SEQ ID NO: 5039)

SNQQ,

(SEQ ID NO: 5040)

RLQQ,

(SEQ ID NO: 5041)

LNQQ,

(SEQ ID NO: 5042)

QRKL,

(SEQ ID NO: 5043)

LRRQ,

(SEQ ID NO: 5044)

QRLR,

(SEQ ID NO: 5045)

QRRL,

(SEQ ID NO: 5046)

RRLQ,

(SEQ ID NO: 5047)

RLRQ,

(SEQ ID NO: 5048)

SKRQ,

(SEQ ID NO: 5049)

QLYR,

(SEQ ID NO: 5050)

QLTV,

(SEQ ID NO: 5051)

QNKQ,

(SEQ ID NO: 5052)

KNQQ,

(SEQ ID NO: 5053)

QKQQ,

(SEQ ID NO: 5054)

QTQQ,

(SEQ ID NO: 5055)

QNHQ,

(SEQ ID NO: 5056)

QHQQ,

(SEQ ID NO: 5057)

QNQH,

(SEQ ID NO: 5058)

QHRQ,

(SEQ ID NO: 5059)

LTQQ,

(SEQ ID NO: 5060)

QNQW,

(SEQ ID NO: 5061)

QNTH,

(SEQ ID NO: 5062)

RRRQ,

(SEQ ID NO: 5063)

QYQQ,

(SEQ ID NO: 5064)

QNDQ,

(SEQ ID NO: 5065)

QNRH,

(SEQ ID NO: 5066)

RDQQ,

(SEQ ID NO: 5067)

PNLQ,

(SEQ ID NO: 5068)

HVRQ,

(SEQ ID NO: 5069)

PNQH,

(SEQ ID NO: 5070)

HNQQ,

(SEQ ID NO: 5071)

QSQQ,

(SEQ ID NO: 5072)

QPAK,

(SEQ ID NO: 5073)

QNLA,

(SEQ ID NO: 5074)

QNQL,

(SEQ ID NO: 5075)

QGQQ,

(SEQ ID NO: 5076)

LNRQ,

(SEQ ID NO: 5077)

QNPP,

(SEQ ID NO: 5078)

QNLQ,

(SEQ ID NO: 5079)

QDQE,

(SEQ ID NO: 5080)

QDQQ,

(SEQ ID NO: 5081)

HWQQ,

(SEQ ID NO: 5082)

PNQQ,

(SEQ ID NO: 5083)

PEQQ,

(SEQ ID NO: 5084)

QRTM,

(SEQ ID NO: 5085)

LHQH,

(SEQ ID NO: 5086)

QHRI,

(SEQ ID NO: 5087)

QYIH,

(SEQ ID NO: 5088)

QKFE,

(SEQ ID NO: 5089)

QFPS,

(SEQ ID NO: 5090)

QNPL,

(SEQ ID NO: 5091)

QAIK,

(SEQ ID NO: 5092)

QNRQ,

(SEQ ID NO: 5093)

QYQH,

(SEQ ID NO: 5094)

QNPQ,

(SEQ ID NO: 5095)

QHQL,

(SEQ ID NO: 5096)

QSPP,

(SEQ ID NO: 5097)

QAKL,

(SEQ ID NO: 5098)

KSQQ,

(SEQ ID NO: 5099)

QDRP,

(SEQ ID NO: 5100)

QNLG,

(SEQ ID NO: 5101)

QAFH,

(SEQ ID NO: 5102)

QNAQ,

(SEQ ID NO: 5103)

HNQL,

(SEQ ID NO: 5104)

QKLN,

(SEQ ID NO: 5105)

QNVQ,

(SEQ ID NO: 5106)

QAQQ,

(SEQ ID NO: 5107)

QTPP,

(SEQ ID NO: 5108)

QPPA,

(SEQ ID NO: 5109)

QERP,

(SEQ ID NO: 5110)

QDLQ,

(SEQ ID NO: 5111)

QAMH,

(SEQ ID NO: 5112)

QHPS,

(SEQ ID NO: 5113)

PGLQ,

(SEQ ID NO: 5114)

QGIR,

(SEQ ID NO: 5115)

QAPA,

(SEQ ID NO: 5116)

QIPP,

(SEQ ID NO: 5117)

QTQL,

(SEQ ID NO: 5118)

QAPS,

(SEQ ID NO: 5119)

QNTY,

(SEQ ID NO: 5120)

QDKQ,

(SEQ ID NO: 5121)

QNHL,

(SEQ ID NO: 5122)

QIGM,

(SEQ ID NO: 5123)

LNKQ,

(SEQ ID NO: 5124)

PNQL,

(SEQ ID NO: 5125)

QLQQ,

(SEQ ID NO: 5126)

QRMS,

(SEQ ID NO: 5127)

QGIL,

(SEQ ID NO: 5128)

QDRQ,

(SEQ ID NO: 5129)

RDWQ,

(SEQ ID NO: 5130)

QERS,

(SEQ ID NO: 5131)

QNYQ,

(SEQ ID NO: 5132)

QRTC,

(SEQ ID NO: 5133)

QIGH,

(SEQ ID NO: 5134)

QGAI,

(SEQ ID NO: 5135)

QVPP,

(SEQ ID NO: 5136)

QVQQ,

(SEQ ID NO: 5137)

LMRQ,

(SEQ ID NO: 5138)

QYSV,

(SEQ ID NO: 5139)

QAIT,

(SEQ ID NO: 5140)

QKTL,

(SEQ ID NO: 5141)

QLHH,

(SEQ ID NO: 5142)

QNII,

(SEQ ID NO: 5143)

QGHH,

(SEQ ID NO: 5144)

QSKV,

(SEQ ID NO: 5145)

QLPS,

(SEQ ID NO: 5146)

IGKQ,

(SEQ ID NO: 5147)

QAIH,

(SEQ ID NO: 5148)

QHGL,

(SEQ ID NO: 5149)

QFMC,

(SEQ ID NO: 5150)

QNQM,

(SEQ ID NO: 5151)

QHLQ,

(SEQ ID NO: 5152)

QPAR,

(SEQ ID NO: 5153)

QSLQ,

(SEQ ID NO: 5154)

QSQL,

(SEQ ID NO: 5155)

HSQQ,

(SEQ ID NO: 5156)

QMPS,

(SEQ ID NO: 5157)

QGSL,

(SEQ ID NO: 5158)

QVPA,

(SEQ ID NO: 5159)

HYQQ,

(SEQ ID NO: 5160)

QVPS,

(SEQ ID NO: 5161)

RGEQ,

(SEQ ID NO: 5162)

PGQQ,

(SEQ ID NO: 5163)

LEQQ,

(SEQ ID NO: 5164)

QNQS,

(SEQ ID NO: 5165)

QKVI,

(SEQ ID NO: 5166)

QNND,

(SEQ ID NO: 5167)

QSVH,

(SEQ ID NO: 5168)

QPLG,

(SEQ ID NO: 5169)

HNQE,

(SEQ ID NO: 5170)

QIQQ,

(SEQ ID NO: 5171)

QVRN,

(SEQ ID NO: 5172)

PSNQ,

(SEQ ID NO: 5173)

QVGH,

(SEQ ID NO: 5174)

QRDI,

(SEQ ID NO: 5175)

QMPN,

(SEQ ID NO: 5176)

RGLQ,

(SEQ ID NO: 5177)

PSLQ,

(SEQ ID NO: 5178)

QRDQ,

(SEQ ID NO: 5179)

QAKG,

(SEQ ID NO: 5180)

QSAH,

(SEQ ID NO: 5181)

QSTM,

(SEQ ID NO: 5182)

QREM,

(SEQ ID NO: 5183)

QYRA,

(SEQ ID NO: 5184)

QRQQ,

(SEQ ID NO: 5185)

QWQQ,

(SEQ ID NO: 5186)

QRMN,

(SEQ ID NO: 5187)

GDSQ,

(SEQ ID NO: 5188)

QKIS,

(SEQ ID NO: 5189)

PSMQ,

(SEQ ID NO: 5190)

SPRQ,

(SEQ ID NO: 5191)

MEQQ,

(SEQ ID NO: 5192)

QYQN,

(SEQ ID NO: 5193)

QIRQ,

(SEQ ID NO: 5194)

QSVQ,

(SEQ ID NO: 5195)

RSQQ,

(SEQ ID NO: 5196)

QNKL,

(SEQ ID NO: 5197)

QIQH,

(SEQ ID NO: 5198)

PRQQ,

(SEQ ID NO: 5199)

HTQQ,

(SEQ ID NO: 5200)

QRQH,

(SEQ ID NO: 5201)

RNQE,

(SEQ ID NO: 5202)

QSKQ,

(SEQ ID NO: 5203)

QNQP,

(SEQ ID NO: 5204)

QSPQ,

(SEQ ID NO: 5205)

QTRQ,

(SEQ ID NO: 5206)

QNLH,

(SEQ ID NO: 5207)

QNQE,

(SEQ ID NO: 5208)

LNQP,

(SEQ ID NO: 5209)

QNQD,

(SEQ ID NO: 5210)

QNLL,

(SEQ ID NO: 5211)

QLVI,

(SEQ ID NO: 5212)

RTQE,

(SEQ ID NO: 5213)

QTHQ,

(SEQ ID NO: 5214)

QDQH,

(SEQ ID NO: 5215)

QSQH,

(SEQ ID NO: 5216)

VRQQ,

(SEQ ID NO: 5217)

AWQQ,

(SEQ ID NO: 5218)

QSVP,

(SEQ ID NO: 5219)

QNIQ,

(SEQ ID NO: 5220)

LDQQ,

(SEQ ID NO: 5221)

PDQQ,

(SEQ ID NO: 5222)

ESQQ,

(SEQ ID NO: 5223)

QRQL,

(SEQ ID NO: 5224)

QIIV,

(SEQ ID NO: 5225)

QKQS,

(SEQ ID NO: 5226)

QSHQ,

(SEQ ID NO: 5227)

QFVV,

(SEQ ID NO: 5228)

QSQP,

(SEQ ID NO: 5229)

QNEQ,

(SEQ ID NO: 5230)

INQQ,

(SEQ ID NO: 5231)

RNRQ,

(SEQ ID NO: 5232)

RDQK,

(SEQ ID NO: 5233)

QWKR,

(SEQ ID NO: 5234)

ENRQ,

(SEQ ID NO: 5235)

QTQP,

(SEQ ID NO: 5236)

QKQL,

(SEQ ID NO: 5237)

RNQL,

(SEQ ID NO: 5238)

ISIQ,

(SEQ ID NO: 5239)

QTVC,

(SEQ ID NO: 5240)

QQIM,

(SEQ ID NO: 5241)

LNHQ,

(SEQ ID NO: 5242)

QNQA,

(SEQ ID NO: 5243)

QMIH,

(SEQ ID NO: 5244)

RNHQ,

or

(SEQ ID NO: 5245)

QKMN;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, or 3 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  42. The AAV capsid variant of any one of embodiments 39-41, wherein [N1]-[N2]-[N3]-[N4] is or comprises:
- (i) the amino acid sequence of any of SEQ ID NOs: 1800-2241;
- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  43. The AAV capsid variant of any one of embodiments 39-42, wherein [N1]-[N2]-[N3]-[N4] is or comprises GSGSPHSKAQNQQ (SEQ ID NO: 1801).
  
  44. The AAV capsid variant of any one of embodiments 39-42, wherein [N1]-[N2]-[N3]-[N4] is or comprises GHDSPHKSGQNQQ (SEQ ID NO: 1800).
  
  45. The AAV capsid variant of any one of embodiments 39-42, wherein [N1]-[N2]-[N3]-[N4] is or comprises GSGSPHKYGQNQQT (SEQ ID NO: 910).
  
  46. The AAV capsid variant of any one of embodiments 1-45, which comprises an amino acid other than T at position 450 (e.g., S, Y, M, A, C, I, R, L, D, F, V, Q, N, H, E, or G), an amino acid other than I at position 451 (e.g., M, P, E, N, D, S, A, T, G, Q, F, V, L, C, H, R, W, or L), and/or an amino acid other than N at position 452 (e.g., M, E, G, Y, W, T, I, Q, F, V, A, L, I, P, K, R, H, S, D, or S), relative to a reference sequence numbered according to any one of SEQ ID NOs: 36-59, 138, 981, or 982.
  
  47. The AAV capsid variant of any one of embodiments 1-46, which comprises the amino acid T at position 450, the amino acid I at position 451, and/or the amino acid N at position 452, relative to a reference sequence numbered according to any one of SEQ ID NOs: 138, 981, or 982.
  
  48. The AAV capsid variant of any one of embodiments 1-47, which further comprises [N0], wherein [N0] comprises XA XB and XC, and wherein:
- (a) position XA is: T, S, Y, M, A, C, I, R, L, D, F, V, Q, N, H, E, or G;
- (b) position XB is: I, M, P, E, N, D, S, A, T, G, Q, F, V, L, C, H, R, W, or L; and
- (c) position XC is: N, M, E, G, Y, W, T, I, Q, F, V, A, L, I, P, K, R, H, S, D, or S; and optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (a)-(c).
  
  49. The AAV capsid variant of embodiment 48, wherein [N0] is or comprises TIN, SMN, TIM, YLS, GLS, MPE, MEG, MEY, AEW, CEW, ANN, IPE, ADM, IEY, ADY, IET, MEW, CEY, RIN, MEI, LEY, ADW, IEI, DIM, FEQ, MEF, CDQ, LPE, IEN, MES, AEI, VEY, IIN, TSN, IEV, MEM, AEV, MDA, VEW, AEQ, LEW, MEL, MET, MEA, IES, MEV, CEI, ATN, MDG, QEV, ADQ, NMN, IEM, ISN, TGN, QQQ, HDW, IEG, TII, TFP, TEK, EIN, TVN, TFN, SIN, TER, TSY, ELH, AIN, SVN, TDN, TFH, TVH, TEN, TSS, TID, TCN, NIN, TEH, AEM, AIK, TDK, TFK, SDQ, TEI, NTN, TET, SIK, TEL, TEA, TAN, TIY, TFS, TES, TTN, TED, TNN, EVH, TIS, TVR, TDR, TIK, NHI, TIP, ESD, TDL, TVP, TVI, AEH, NCL, TVK, NAD, TIT, NCV, TIR, NAL, VIN, TIQ, TEF, TRE, QGE, SEK, NVN, GGE, EFV, SDK, TEQ, EVQ, TEY, NCW, TDV, SDI, NSI, NSL, EVV, TEP, SEL, TWQ, TEV, AVN, GVL, TLN, TEG, TRD, NAI, AEN, AET, ETA, NNL, or any dipeptide thereof.
  
  50. The AAV capsid variant of embodiment 48 or 49, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or comprises:
- (i) the amino acid sequence of any one of SEQ ID NOs: 2242-2886;
- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  51. The AAV capsid variant of any one of embodiments 48-50, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or comprises TINGSGSPHSKAQNQQ (SEQ ID NO: 2242).
  
  52. The AAV capsid variant of any one of embodiments 48-50, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or comprises TINGHDSPHKSGQNQQ (SEQ ID NO: 2243).
  
  53. The AAV capsid variant of any one of embodiments 48-52, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or comprises TINGSGSPHKYGQNQQT (SEQ ID NO: 5246).
  
  54. The AAV capsid variant of any one of embodiments 1-53, wherein [N1]-[N2]-[N3] is present in loop IV.
  
  55. The AAV capsid variant of any one of embodiments 48-54, wherein [N0] and [N4] are present in loop IV.
  
  56. The AAV capsid variant of any one of embodiments 48-55, wherein [N0] is present immediately subsequent to position 449, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  57. The AAV capsid variant of any one of embodiments 48-56, wherein [N0] is present immediately subsequent to position 449, relative to a reference sequence numbered according to the amino acid sequence of any one of SEQ ID NOs: 36-59, 981, or 982.
  
  58. The AAV capsid variant of any one of embodiments 48-57, wherein [N0] replaces positions 450, 451, and 452 (e.g., T450, I451, and N452), relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  59. The AAV capsid variant of any one of embodiments 48-58, wherein [N0] replaces positions 450-452 (e.g., T450, 1451, and N452), relative to a reference sequence numbered according to any one of SEQ ID NOs: 36-59, 981, or 982.
  
  60. The AAV capsid variant of any one of embodiments 48-59, wherein [N0] corresponds to positions 450-452 of any one of SEQ ID NOs: 36-59, 138, 981 or 982.
  
  61. The AAV capsid variant of any one of embodiments 48-60, wherein [N0] is present immediately subsequent to position 449 and wherein [N0] replaces positions 450-452 (e.g., T450, 1451, and N452), relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  62. The AAV capsid variant of any one of embodiments 48-61, wherein [N0] is present immediately subsequent to position 449 and wherein [N0] replaces positions 450-452 (e.g., T450, 1451, and N452), relative to a reference sequence numbered according to any one of SEQ ID NOs: 36-59, 981, or 982.
  
  63. The AAV capsid variant of any one of embodiments 1-62, wherein [N1] is present immediately subsequent to position 452, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  64. The AAV capsid variant of any one of embodiments 1-63, wherein [N1] is present immediately subsequent to position 452, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981 or 982.
  
  65. The AAV capsid variant of any one of embodiments 1-61, wherein [N1] replaces positions 453-455 (e.g., G453, S454, and G455), relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  66. The AAV capsid variant of any one of embodiments 1-64, wherein [N1] replaces positions 453 (e.g., G453), relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  67. The AAV capsid variant of any one of embodiments 1-65, wherein [N1] replaces positions 453-455 (e.g., G453, S454, and G455), relative to a reference sequence numbered according to SEQ ID NO: 981.
  
  68. The AAV capsid variant of any one of embodiments 1-65 or 67, wherein [N1] replaces positions 453-455, relative to a reference sequence numbered according to SEQ ID NO: 982.
  
  69. The AAV capsid variant of any one of embodiments 1-65, 67, or 68, wherein [N1] is present immediately subsequent to position 452 and wherein [N1] replaces positions 453-455 (e.g., G453, S454, and G455), relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  70. The AAV capsid variant of any one of embodiments 1-64 or 66, wherein [N1] is present immediately subsequent to position 452 and wherein [N1] replaces positions 453 (e.g., G453), relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  71. The AAV capsid variant of any one of embodiments 1-64, 66 or 70, wherein [N1] is present immediately subsequent to position 452 and wherein [N1] replaces positions 453-455, relative to a reference sequence numbered according to SEQ ID NO: 981 or 982.
  
  72. The AAV capsid variant of any one of embodiments 1-71, wherein [N1] corresponds to positions 453-455 of any one of SEQ ID NOs: 36-59, 981 or 982.
  
  73. The AAV capsid variant of any one of embodiment 1-72, which comprises an amino acid other than S at position 454 and/or an amino acid other than G at position 455, numbered according to SEQ ID NO: 138, 981, or 982.
  
  74. The AAV capsid variant of any one of embodiments 1-73, which comprises the amino acid H at position 454 and the amino acid D at position 455, numbered according to SEQ ID NO: 138 or 982.
  
  75. The AAV capsid variant of any one of embodiments 1-74, which comprises a substitution at position 454 (e.g., S454H) and/or a substitution at position 455 (e.g., G455D), numbered according to SEQ ID NO: 138.
  
  76. The AAV capsid variant of any one of embodiments 1-75, which comprises the amino acid H at position 454 and the amino acid D at position 455, and further comprises the amino acid sequence SPHSKA (SEQ ID NO: 941) immediately subsequent to position 455, relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  77. The AAV capsid variant of any one of embodiments 1-76, which comprises the amino acid H at position 454 and the amino acid D at position 455, relative to a reference sequence numbered according to SEQ ID NO: 982.
  
  78. The AAV capsid variant of any one of embodiments 1-77, which comprises the amino acid H at position 454 and the amino acid D at position 455, and further comprises the amino acid sequence SPHSKA (SEQ ID NO: 941) immediately subsequent to position 455, relative to a reference sequence numbered according to SEQ ID NO: 982.
  
  79. The AAV capsid variant of any one of embodiments 1-72, which comprises the amino acid S at position 454 and the amino acid G at position 455, relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  80. The AAV capsid variant of any one of embodiments 1-72 or 79, which comprises the amino acid S at position 454 and the amino acid G at position 455, and further comprises the amino acid sequence SPHSKA (SEQ ID NO: 941) immediately subsequent to position 455, relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  81. The AAV capsid variant of any one of embodiments 1-72, 79, or 80, which comprises the amino acid S at position 454 and the amino acid G at position 455, relative to a reference sequence numbered according to SEQ ID NO: 981.
  
  82. The AAV capsid variant of any one of embodiments 1-72 or 79-81, which comprises the amino acid S at position 454 and the amino acid G at position 455, and further comprises the amino acid sequence SPHSKA (SEQ ID NO: 941) immediately subsequent to position 455, relative to a reference sequence numbered according to SEQ ID NO: 981.
  
  83. The AAV capsid variant of any one of embodiments 1-82, wherein [N2] is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  84. The AAV capsid variant of any one of embodiments 1-83, wherein [N2] corresponds to positions 456-458 (e.g., S456, P457, H458) of SEQ ID NO: 981 or 982.
  
  85. The AAV capsid variant of any one of embodiments 1-83, wherein [N2] corresponds to positions 456-458 (e.g., S456, P457, H458) of any one of SEQ ID NOs: 36-59.
  
  86. The AAV capsid variant of any one of embodiments 1-85, wherein [N2]-[N3] is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  87. The AAV capsid variant of any one of embodiments 1-86, wherein [N2] is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981 or 982.
  
  88. The AAV capsid variant of any one of embodiments 1-87, wherein [N2]-[N3] is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981 or 982.
  
  89. The AAV capsid variant of any one of embodiments 1-88, wherein [N2]-[N3] corresponds to positions 456-461 (e.g., S456, P457, H458, S459, K460, A461) of SEQ ID NO: 981.
  
  90. The AAV capsid variant of any one of embodiments 1-88, wherein [N2]-[N3] corresponds to positions 456-461 (e.g., S456, P457, H458, K459, S460, G461) of SEQ ID NO: 982.
  
  91. The AAV capsid variant of any one of embodiments 1-90, wherein [N2] is present immediately subsequent to [N1].
  
  92. The AAV capsid variant of any one of embodiments 1-64, 66, 70, or 71, wherein [N3] is present immediately subsequent to [N2] and replaces positions 454 and 455 (e.g., S454 and G455), numbered according to SEQ ID NO: 138.
  
  93. The AAV capsid variant of any one of embodiments 1-1-64, 66, 70, 71, or 92, wherein [N3] is present immediately subsequent to [N1]-[N2] and replaces positions 454 and 455 (e.g., S454 and G455), numbered according to SEQ ID NO: 138.
  
  94. The AAV capsid variant of any one of embodiments 39-93, wherein [N4] is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  95. The AAV capsid variant of any one of embodiments 39-94, wherein [N4] replaces positions 456-459 (e.g., Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  96. The AAV capsid variant of any one of embodiments 39-95, wherein [N4] corresponds to positions 462-465 (e.g., Q462, N463, Q464, Q465) of SEQ ID NO: 981 or 982.
  
  97. The AAV capsid variant of any one of embodiments 39-96, wherein [N2]-[N3]-[N4] replaces positions 456-459 (e.g., Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  98. The AAV capsid variant of any one of embodiments 39-97, wherein [N2]-[N3]-[N4] is present immediately subsequent to position 455, and wherein [N2]-[N3]-[N4] replaces positions 456-459 (e.g., Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  99. The AAV capsid variant of any one of embodiments 39-98, wherein [N2]-[N3]-[N4] corresponds to positions 456-465 (e.g., S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465) of SEQ ID NO: 981.
  
  100. The AAV capsid variant of any one of embodiments 39-98, wherein [N2]-[N3]-[N4] corresponds to positions 456-465 (e.g., S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465) of SEQ ID NO: 982.
  
  101. The AAV capsid variant of any one of embodiments 39-98, wherein [N2]-[N3]-[N4] corresponds to positions 456-465 of any one of SEQ ID NOs: 36-59.
  
  102. The AAV capsid variant of any one of embodiments 39-101, wherein [N1]-[N2]-[N3]-[N4] replaces positions 453-459 (e.g., G453, S454, G455, Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  103. The AAV capsid variant of any one of embodiments 39-102, wherein [N1]-[N2]-[N3]-[N4] is present immediately subsequent to position 452, and wherein [N1]-[N2]-[N3]-[N4] replaces positions 453-459 (e.g., G453, S454, G455, Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  104. The AAV capsid variant of any one of embodiments 39-99, 102,103, wherein [N1]-[N2]-[N3]-[N4] corresponds to positions 453-465 (e.g., G453, S454, G455, S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465) of SEQ ID NO: 981.
  
  105. The AAV capsid variant of any one of embodiments 39-98, 100, 102, or103, wherein [N1]-[N2]-[N3]-[N4] corresponds to positions 453-465 (e.g., G453, H454, D455, S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465) of SEQ ID NO: 982.
  
  106. The AAV capsid variant of any one of embodiments 39-98, 102, or 103, wherein [N1]-[N2]-[N3]-[N4] corresponds to positions 453-465 of any one of SEQ ID NOs: 36-59.
  
  107. The AAV capsid variant of any one of embodiments 1-99 or 102-104, wherein [N1]-[N2]-[N3] corresponds to positions 453-461 (e.g., G453, S454, G455, S456, P457, H458, S459, K460, A461) of SEQ ID NO: 981.
  
  108. The AAV capsid variant of any one of embodiments 1-98, 100, 102, 103, or 105, wherein [N1]-[N2]-[N3] corresponds to positions 453-461 (e.g., G453, H454, D455, S456, P457, H458, K459, S460, G461) of SEQ ID NO: 982.
  
  109. The AAV capsid variant of any one of embodiments 39-98, 102, 103, or 106, wherein [N1]-[N2]-[N3] corresponds to positions 453-461 of any one of SEQ ID NOs: 36-59.
  
  110. The AAV capsid variant of any one of embodiments 48-109, wherein [N0]-[N1]-[N2]-[N3]-[N4] replaces positions 450-459 (e.g., T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  111. The AAV capsid variant of any one of embodiments 48-110, wherein [N0]-[N1]-[N2]-[N3]-[N4] is present immediately subsequent to position 449, and wherein [N0]-[N1]-[N2]-[N3]-[N4] replaces positions 450-459 (e.g., T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  112. The AAV capsid variant of any one of embodiments 48-99, 102-104, or 106, wherein [N0]-[N1]-[N2]-[N3]-[N4] corresponds to positions 450-465 (e.g., T450, 1451, N452, G453, S454, G455, S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465) of SEQ ID NO: 981.
  
  113. The AAV capsid variant of any one of embodiments 48-98, 100, 102, 103, 105, or 108, wherein [N0]-[N1]-[N2]-[N3]-[N4] corresponds to positions 450-465 (e.g., T450, 1451, N452, G453, H454, D455, S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465) of SEQ ID NO: 982.
  
  114. The AAV capsid variant of any one of embodiments 48-98, 102, 103, 106, or 109, wherein [N0]-[N1]-[N2]-[N3]-[N4] corresponds to positions 450-465 of any one of SEQ ID NOs: 36-59.
  
  115. The AAV capsid variant of any one of embodiments 39-114, wherein [N4] replaces positions 462-465 (e.g., Q462, N463, Q464, and Q465), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981 or 982.
  
  116. The AAV capsid variant of any one of embodiments 39-115, wherein [N2]-[N3]-[N4] replaces positions 462-465 (e.g., Q462, N463, Q464, and Q465), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981 or 982.
  
  117. The AAV capsid variant of any one of embodiments 39-116, wherein [N2]-[N3]-[N4] is present immediately subsequent to position 455, and wherein [N2]-[N3]-[N4] replaces positions 462-465 (e.g., Q462, N463, Q464, and Q465), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981 or 982.
  
  118. The AAV capsid variant of any one of embodiments 1-117, wherein [N3] is present immediately subsequent to [N2].
  
  119. The AAV capsid variant of any one of embodiments 1-118, which comprises from N-terminus to C-terminus, [N2]-[N3].
  
  120. The AAV capsid variant of any one of embodiments 1-119, which comprises from N-terminus to C-terminus, [N1]-[N2]-[N3].
  
  121. The AAV capsid variant of any one of embodiments 48-120, which comprises from N-terminus to C-terminus, [N0]-[N1]-[N2]-[N3].
  
  122. The AAV capsid variant of any one of embodiments 39-121, which comprises from N-terminus to C-terminus, [N1]-[N2]-[N3]-[N4].
  
  123. The AAV capsid variant of any one of embodiments 48-122, which comprises from N-terminus to C-terminus, [N0]-[N1]-[N2]-[N3]-[N4].
  
  124. The AAV capsid variant of any one of the preceding embodiments, which comprises an amino acid other T at position 460 (e.g., N, I, C, H, R, L, D, Y, A, M, Q, I, E, K, P, G or S), numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  125. The AAV capsid variant of any one of the preceding embodiments, which comprises the amino acid N, I, C, H, R, L, D, Y, A, M, Q, I, E, K, P, G or S at position 460, numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  126. The AAV capsid variant of any one of the preceding embodiments, which comprises an amino acid other T at position 466 (e.g., N, I, C, H, R, L, D, Y, A, M, Q, I, E, K, P, G or S), numbered according to the amino acid sequence of any one of SEQ ID NOs: 36-59, 981, or 982.
  
  127. The AAV capsid variant of any one of the preceding embodiments, which comprises the amino acid N, I, C, H, R, L, D, Y, A, M, Q, I, E, K, P, G or S at position 466, numbered according to the amino acid sequence of any one of SEQ ID NOs: 36-59, 981 or 982.
  
  128. The AAV capsid variant of any one of the preceding embodiments, which comprises an amino acid other K at position 449 (e.g., an E, an N, or a T), numbered according to the amino acid sequence of any one of SEQ ID NOs: 36-59, 138, 981, or 982.
  
  129. The AAV capsid variant of any one of the preceding embodiments, which comprises the amino E, N, or T at position 449, numbered according to the amino acid sequence of any one of SEQ ID NOs: 36-59, 138, 981 or 982.
  
  130. An AAV capsid variant, comprising [A][B](SEQ ID NO: 4694), wherein:
- (i) [A] comprises the amino acid sequence of GSGSPH (SEQ ID NO: 4695); and
- (ii) [B] comprises X1 X2 X3 X4 X5 X6 X7, wherein:
  - (a) position X1 is: S, C, F, or V;
  - (b) position X2 is: K, L, R, I, E, Y, V, or S;
  - (c) position X3 is: A, R, L, G, I, Y, S, F, or W;
  - (d) position X4 is: W, Q, R, G, L, V, S, or F;
  - (e) position X5 is: N, Y, R, C, K, or L;
  - (f) position X6 is: Q, G, K, R, T, L, or Y; and
  - (g) position X7 is: Q, L, R, or V;
- optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (a)-(g).
  
  131. The AAV capsid variant of embodiment 130, wherein
- (a) position X1 is S;
- (b) position X2 is K or L;
- (c) position X3 is: A, R, or L;
- (d) position X4 is: Q or R;
- (e) position X5 is: N or R;
- (f) position X6 is: Q or R; and
- (g) position X7 is: Q, L, or R.
  
  132. The AAV capsid variant of embodiment 130 or 131, wherein [B] comprises:

(i)

(SEQ ID NO: 5247)

SLLWNQQ,

(SEQ ID NO: 5248)

SKAQYYV,

(SEQ ID NO: 5249)

SKLRRQQ,

(SEQ ID NO: 5250)

SIWQNQQ,

(SEQ ID NO: 5251)

SKAGCGQ,

(SEQ ID NO: 5252)

SRAQNQQ,

(SEQ ID NO: 5253)

SKRLRQQ,

(SEQ ID NO: 5254)

SLRRNQQ,

(SEQ ID NO: 5255)

SRGRNQQ,

(SEQ ID NO: 5256)

SEIVNQQ,

(SEQ ID NO: 5257)

SSRRNQQ,

(SEQ ID NO: 5258)

CLLQNQQ,

(SEQ ID NO: 5259)

SKAFRLQ,

(SEQ ID NO: 5260)

CLAQNQQ,

(SEQ ID NO: 5261)

FLRQNQQ,

(SEQ ID NO: 5262)

SLRFNQQ,

(SEQ ID NO: 5263)

SYLRNQQ,

(SEQ ID NO: 5264)

CSLQNQQ,

(SEQ ID NO: 5265)

VLWQNQQ,

(SEQ ID NO: 5266)

SKWLLQQ,

(SEQ ID NO: 5267)

SLWSNQQ,

(SEQ ID NO: 5268)

SKRRLQQ,

(SEQ ID NO: 5269)

SVYLNQQ,

(SEQ ID NO: 5270)

SLWLNQQ,

(SEQ ID NO: 5271)

SKAQRKL,

(SEQ ID NO: 5272)

SKALRRQ,

(SEQ ID NO: 5273)

SKAQRLR,

(SEQ ID NO: 5274)

SKAQNQQ,

(SEQ ID NO: 5275)

SKAQRRL,

(SEQ ID NO: 5276)

SKARRQQ,

(SEQ ID NO: 5277)

SKARRLQ,

(SEQ ID NO: 5278)

SKSRRQQ,

(SEQ ID NO: 5279)

SKARLRQ,

(SEQ ID NO: 5280)

SKASKRQ,

(SEQ ID NO: 5281)

VRRQNQQ,

(SEQ ID NO: 5282)

SKAQLYR,

(SEQ ID NO: 5283)

SLFRNQQ,

(SEQ ID NO: 5284)

SKAQLTV;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, or 6 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  133. The AAV capsid variant of any one of embodiments 130-132, wherein [A][B] comprises:

(i)

(SEQ ID NO: 5285)

GSGSPHSLLWNQQ,

(SEQ ID NO: 2060)

GSGSPHSKAQYYV,

(SEQ ID NO: 2061)

GSGSPHSKLRRQQ,

(SEQ ID NO: 5286)

GSGSPHSIWQNQQ,

(SEQ ID NO: 2062)

GSGSPHSKAGCGQ,

(SEQ ID NO: 2063)

GSGSPHSRAQNQQ,

(SEQ ID NO: 2064)

GSGSPHSKRLRQQ,

(SEQ ID NO: 2065)

GSGSPHSLRRNQQ,

(SEQ ID NO: 2066)

GSGSPHSRGRNQQ,

(SEQ ID NO: 5287)

GSGSPHSEIVNQQ,

(SEQ ID NO: 2067)

GSGSPHSSRRNQQ,

(SEQ ID NO: 5288)

GSGSPHCLLQNQQ,

(SEQ ID NO: 2068)

GSGSPHSKAFRLQ,

(SEQ ID NO: 5289)

GSGSPHCLAQNQQ,

(SEQ ID NO: 2070)

GSGSPHFLRQNQQ,

(SEQ ID NO: 2071)

GSGSPHSLRFNQQ,

(SEQ ID NO: 5290)

GSGSPHSYLRNQQ,

(SEQ ID NO: 5291)

GSGSPHCSLQNQQ,

(SEQ ID NO: 5292)

GSGSPHVLWQNQQ,

(SEQ ID NO: 2072)

GSGSPHSKWLLQQ,

(SEQ ID NO: 5293)

GSGSPHSLWSNQQ,

(SEQ ID NO: 2073)

GSGSPHSKRRLQQ,

(SEQ ID NO: 5294)

GSGSPHSVYLNQQ,

(SEQ ID NO: 5295)

GSGSPHSLWLNQQ,

(SEQ ID NO: 2074)

GSGSPHSKAQRKL,

(SEQ ID NO: 2075)

GSGSPHSKALRRQ,

(SEQ ID NO: 2076)

GSGSPHSKAQRLR,

(SEQ ID NO: 1801)

GSGSPHSKAQNQQ,

(SEQ ID NO: 2077)

GSGSPHSKAQRRL,

(SEQ ID NO: 2078)

GSGSPHSKARRQQ,

(SEQ ID NO: 2079)

GSGSPHSKARRLQ,

(SEQ ID NO: 2080)

GSGSPHSKSRRQQ,

(SEQ ID NO: 2082)

GSGSPHSKARLRQ,

(SEQ ID NO: 2083)

GSGSPHSKASKRQ,

(SEQ ID NO: 2084)

GSGSPHVRRQNQQ,

(SEQ ID NO: 2085)

GSGSPHSKAQLYR,

(SEQ ID NO: 5296)

GSGSPHSLFRNQQ,

(SEQ ID NO: 2086)

GSGSPHSKAQLTV;.

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i)
  
  134. The AAV capsid variant of any one of embodiments 130-133, which further comprises one, two, or all of an amino acid other than T at position 450 (e.g., S, Y, or G), an amino acid other than I at position 451 (e.g., M or L), and/or an amino acid other than N at position 452 (e.g., S), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  135. The AAV capsid variant of any one of embodiments 130-134, which further comprises an S at position 450 and an M at position 451, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  136. The AAV capsid variant of any one of embodiments 130-134, which further comprises a Y at position 450, an L at position 451, and an S at position 452, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  137. The AAV capsid variant of any one of embodiments 130-134, which further comprises a G at position 450, an L at position 451, and an S at position 452, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  138. The AAV capsid variant of any one of embodiments 130-137, wherein [A][B] is present in loop IV.
  
  139. The AAV capsid variant of any one of embodiments 130-138, wherein [A] is present immediately subsequent to position 452, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  140. The AAV capsid variant of any one of embodiments 130-139, wherein [A] replaces positions 453-455 (e.g., G453, S454, G455), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  141. The AAV capsid variant of any one of embodiments 130-140, wherein [A] is present immediately subsequent to position 452, and wherein [A] replaces positions 453-455 (e.g., G453, S454, G455), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  142. The AAV capsid variant of any one of embodiments 130-141, wherein [B] is present immediately subsequent to [A].
  
  143. The AAV capsid variant of any one of embodiments 130-142, wherein [B] replaces positions 456-459 (e.g., Q456, N457, Q458, Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  144. The AAV capsid variant of any one of embodiments 130-143, wherein [A][B] replaces positions 453-459 (e.g., G453, S454, G455, Q456, N457, Q458, Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  145. The AAV capsid variant of any one of embodiments 130-144, wherein [A][B] is present immediately subsequent to position 452, and wherein [A][B] replaces positions 453-459 (e.g., G453, S454, G455, Q456, N457, Q458, Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  146. The AAV capsid variant of any one of embodiments 130-145, which comprises from N-terminus to C-terminus, [A][B].
  
  147. An AAV capsid comprising [A][B](SEQ ID NO: 4699), wherein:
- (i) [A] comprises X1 X2 X3 X4 X5 X6, wherein
  - (a) position X1 is T, M, A, C, I, R, L, D, F, V, Q, N, or H;
  - (b) position X2 is I, P, E, N, D, S, A, T, M, or Q;
  - (c) position X3 is N, E, G, Y, W, M, T, I, K, Q, F, S, V, A, or L;
  - (d) position X4 is G, D, R, or E;
  - (e) position X5 is H, Q, N, or D;
  - (f) position X6 is D or R;
- optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (a)-(f); and
- (ii) [B] comprises SPHKSG (SEQ ID NO: 946).
  
  148. The AAV capsid of embodiment 147, wherein
- (a) position X1 is: T, M, A, or I;
- (b) position X2 is: E, I or D;
- (c) position X3 is: N, Q, Y, I, M, or V;
- (d) position X4 is G;
- (e) position X5 is H; and
- (f) position X6 is D.
  
  149. The AAV capsid variant of embodiment 147 or 148, wherein [A] comprises:

(i)

(SEQ ID NO: 5297)

TINGHD,

(SEQ ID NO: 5298)

MPEGHD,

(SEQ ID NO: 5299)

MEGGHD,

(SEQ ID NO: 5300)

MEYGHD,

(SEQ ID NO: 5301)

AEWGHD,

(SEQ ID NO: 5302)

CEWGHD,

(SEQ ID NO: 5303)

ANNGQD,

(SEQ ID NO: 5304)

IPEGHD,

(SEQ ID NO: 5305)

ADMGHD,

(SEQ ID NO: 5306)

IEYGHD,

(SEQ ID NO: 5307)

ADYGHD,

(SEQ ID NO: 5308)

IETGHD,

(SEQ ID NO: 5309)

MEWGHD,

(SEQ ID NO: 5310)

CEYGHD,

(SEQ ID NO: 5311)

RINGHD,

(SEQ ID NO: 5312)

MEIGHD,

(SEQ ID NO: 5313)

LEYGHD,

(SEQ ID NO: 5314)

ADWGHD,

(SEQ ID NO: 5315)

IEIGHD,

(SEQ ID NO: 5316)

TIKDND,

(SEQ ID NO: 5317)

DIMGHD,

(SEQ ID NO: 5318)

FEQGHD,

(SEQ ID NO: 5319)

MEFGHD,

(SEQ ID NO: 5320)

CDQGHD,

(SEQ ID NO: 5321)

LPEGHD,

(SEQ ID NO: 5322)

IENGHD,

(SEQ ID NO: 5323)

MESGHD,

(SEQ ID NO: 5324)

AEIGHD,

(SEQ ID NO: 5325)

VEYGHD,

(SEQ ID NO: 5326)

TSNGDD,

(SEQ ID NO: 5327)

IEVGHD,

(SEQ ID NO: 5328)

MEMGHD,

(SEQ ID NO: 5329)

AEVGHD,

(SEQ ID NO: 5330)

MDAGHD,

(SEQ ID NO: 5331)

VEWGHD,

(SEQ ID NO: 5332)

AEQGHD,

(SEQ ID NO: 5333)

LEWGHD,

(SEQ ID NO: 5334)

MELGHD,

(SEQ ID NO: 5335)

METGHD,

(SEQ ID NO: 5336)

MEAGHD,

(SEQ ID NO: 5337)

TINRQR,

(SEQ ID NO: 5338)

IESGHD,

(SEQ ID NO: 5339)

TAKDHD,

(SEQ ID NO: 5340)

MEVGHD,

(SEQ ID NO: 5341)

CEIGHD,

(SEQ ID NO: 5342)

ATNGHD,

(SEQ ID NO: 5343)

MDGGHD,

(SEQ ID NO: 5344)

QEVGHD,

(SEQ ID NO: 5345)

ADQGHD,

(SEQ ID NO: 5346)

NMNGHD,

(SEQ ID NO: 5347)

TPWEHD,

(SEQ ID NO: 5348)

IEMGHD,

(SEQ ID NO: 5349)

TANEHD,

(SEQ ID NO: 5350)

QQQGHD,

(SEQ ID NO: 5351)

TPQDHD,

(SEQ ID NO: 5352)

HDWGHD,

(SEQ ID NO: 5353)

IEGGHD,

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, or 5 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  150. The AAV capsid variant of any one of embodiments 147-149, wherein [A][B] comprises:

(i)

(SEQ ID NO: 5354)

TINGHDSPHKR,

(SEQ ID NO: 5355)

MPEGHDSPHKS,

(SEQ ID NO: 5356)

MEGGHDSPHKS,

(SEQ ID NO: 5357)

MEYGHDSPHKS,

(SEQ ID NO: 5358)

AEWGHDSPHKS,

(SEQ ID NO: 5359)

CEWGHDSPHKS,

(SEQ ID NO: 5360)

ANNGQDSPHKS,

(SEQ ID NO: 5361)

IPEGHDSPHKS,

(SEQ ID NO: 5362)

ADMGHDSPHKS,

(SEQ ID NO: 5363)

IEYGHDSPHKS,

(SEQ ID NO: 5364)

ADYGHDSPHKS,

(SEQ ID NO: 5365)

IETGHDSPHKS,

(SEQ ID NO: 5366)

MEWGHDSPHKS,

(SEQ ID NO: 5367)

CEYGHDSPHKS,

(SEQ ID NO: 5368)

RINGHDSPHKS,

(SEQ ID NO: 5369)

MEIGHDSPHKS,

(SEQ ID NO: 5370)

LEYGHDSPHKS,

(SEQ ID NO: 5371)

ADWGHDSPHKS,

(SEQ ID NO: 5372)

IEIGHDSPHKS,

(SEQ ID NO: 5373)

TIKDNDSPHKS,

(SEQ ID NO: 5374)

DIMGHDSPHKS,

(SEQ ID NO: 5375)

FEQGHDSPHKS,

(SEQ ID NO: 5376)

MEFGHDSPHKS,

(SEQ ID NO: 5377)

CDQGHDSPHKS,

(SEQ ID NO: 5378)

LPEGHDSPHKS,

(SEQ ID NO: 5379)

IENGHDSPHKS,

(SEQ ID NO: 5380)

MESGHDSPHKS,

(SEQ ID NO: 5381)

AEIGHDSPHKS,

(SEQ ID NO: 5382)

VEYGHDSPHKS,

(SEQ ID NO: 5383)

TSNGDDSPHKS,

(SEQ ID NO: 5384)

IEVGHDSPHKS,

(SEQ ID NO: 5385)

MEMGHDSPHKS,

(SEQ ID NO: 5386)

AEVGHDSPHKS,

(SEQ ID NO: 5387)

MDAGHDSPHKS,

(SEQ ID NO: 5388)

VEWGHDSPHKS,

(SEQ ID NO: 5389)

AEQGHDSPHKS,

(SEQ ID NO: 5390)

LEWGHDSPHKS,

(SEQ ID NO: 5391)

MELGHDSPHKS,

(SEQ ID NO: 5392)

METGHDSPHKS,

(SEQ ID NO: 5393)

MEAGHDSPHKS,

(SEQ ID NO: 5394)

TINRQRSPHKS,

(SEQ ID NO: 5395)

IESGHDSPHKS,

(SEQ ID NO: 5396)

TAKDHDSPHKS,

(SEQ ID NO: 5397)

MEVGHDSPHKS,

(SEQ ID NO: 5398)

CEIGHDSPHKS,

(SEQ ID NO: 5399)

ATNGHDSPHKS,

(SEQ ID NO: 5400)

MDGGHDSPHKS,

(SEQ ID NO: 5401)

QEVGHDSPHKS,

(SEQ ID NO: 5402)

ADQGHDSPHKS,

(SEQ ID NO: 5403)

NMNGHDSPHKS,

(SEQ ID NO: 5404)

TPWEHDSPHKS,

(SEQ ID NO: 5405)

IEMGHDSPHKS,

(SEQ ID NO: 5406)

TANEHDSPHKS,

(SEQ ID NO: 5407)

TINGHDSPHKS,

(SEQ ID NO: 5408)

QQQGHDSPHKS,

(SEQ ID NO: 5409)

TPQDHDSPHKS,

(SEQ ID NO: 5410)

HDWGHDSPHKS,

(SEQ ID NO: 5411)

IEGGHDSPHKS

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  151. The AAV capsid variant of any one of embodiments 147-150, which further comprises one, two, three, four, or all of an amino acid other than Q at position 456 (e.g., R or L), N at position 457 (e.g., H, K, or R), Q at position 458 (e.g., R or T), Q at position 459 (H), and/or T at position 460 (N or S), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  152. The AAV capsid variant of any one of embodiments 147-151, which further comprises an R at position 456, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  153. The AAV capsid variant of any one of embodiments 147-151, which further comprises an L at position 456, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  154. The AAV capsid variant of any one of embodiments 147-153, which further comprises an H at position 457 and an R at position 458, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  155. The AAV capsid variant of any one of embodiments 147-153, which further comprises a K at position 457 and an N at position 460, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  156. The AAV capsid variant of any one of embodiments 147-153, which further comprises a T at position 458, an H at position 459, and an S at position 460, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  157. The AAV capsid variant of any one of embodiments 147-151, which further comprises an R at position 456, an R at position 457, and an R at position 458, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  158. The AAV capsid variant of any one of embodiments 147-157, wherein [A][B] is present in loop IV.
  
  159. The AAV capsid variant of any one of embodiments 147-158, wherein [A] is present immediately subsequent to position 449, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  160. The AAV capsid variant of any one of embodiments 147-159, wherein [A] replaces positions 450-453 (e.g., T450, 1451, N452, G453), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  161. The AAV capsid variant of any one of embodiments 147-160, wherein [A] is present immediately subsequent to position 449, and wherein [A] replaces positions 450-453 (e.g., T450, 1451, N452, G453), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  162. The AAV capsid variant of any one of embodiments 147-161, wherein [A][B] replaces positions 450-455 (e.g., T450, 1451, N452, G453, S454, G455), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  163. The AAV capsid variant of any one of embodiments 147-162, wherein [A][B] is present immediately subsequent to position 449, and wherein [A][B] replaces positions 450-455 (e.g., T450, I451, N452, G453, S454, G455), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  164. The AAV capsid variant of any one of embodiments 147-163, wherein [B] is present immediately subsequent [A], and replaces positions 454 and 455 (e.g., S454 and G455), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  165. The AAV capsid variant of any one of embodiments 147-164, wherein [B] is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 982.
  
  166. The AAV capsid variant of any one of embodiments 147-165, wherein [B] is present immediately subsequent to [A].
  
  167. The AAV capsid variant of any one of embodiments 147-166, which comprises from N-terminus to C-terminus, [A][B].
  
  168. An AAV capsid variant comprising an amino acid sequence having the following formula: [N1]-[N2]-[N3](SEQ ID NO: 6407), wherein:
- (i) [N1] comprises positions X1, X2, and X3, wherein position X2 is S and position X3 is G;
- (ii) [N2] comprises the amino acid sequence SPH; and
- (iii) [N3] comprises positions X4, X5, and X6, wherein position X5 is K.
  
  169. The AAV capsid variant of embodiment 168, wherein:
- (i) X4 of [N3] is S, T, N, or A; and
- (ii) X5 of [N3] is A, V, T, S, G, R, L, or N;
- optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (i) or (ii).
  
  170. The AAV capsid variant of embodiment 168 or 169, wherein X4 is S and/or X5 is A.
  
  171. The AAV capsid variant of any one of embodiments 168-170, wherein [N3] comprises SK, TK, NK, AK, KA, KV, KT, KS, KG, KR, KL, or KN.
  
  172. The AAV capsid variant of any one of embodiments 168-171, wherein [N3] is or comprises SKA, SKV, SKT, SKS, SKG, SKR, TKA, NKA, SKL, SKN, or AKA.
  
  173. The AAV capsid variant of any one of embodiments 168-172, wherein [N3] is or comprises SKA.
  
  174. The AAV capsid variant of any one of embodiments 168-173, wherein [N2]-[N3] comprises SPHSK (SEQ ID NO: 4701), SPHTK (SEQ ID NO: 4725), SPHNK (SEQ ID NO: 4726), or SPHAK (SEQ ID NO: 4727).
  
  175. The AAV capsid variant of any one of embodiments 168-174, wherein [N2]-[N3] is or comprises:

(i)

(SEQ ID NO: 941)

SPHSKA,

(SEQ ID NO: 4737)

SPHSKV,

(SEQ ID NO: 4731)

SPHSKT,

(SEQ ID NO: 962)

SPHSKS,

(SEQ ID NO: 4732)

SPHSKG,

(SEQ ID NO: 978)

SPHSKR,

(SEQ ID NO: 4739)

SPHTKA,

(SEQ ID NO: 4734)

SPHNKA,

(SEQ ID NO: 960)

SPHSKL,

(SEQ ID NO: 4735)

SPHSKN,

or

(SEQ ID NO: 4736)

SPHAKA;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, or 5 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  176. The AAV capsid variant of any one of embodiments 168-175, wherein [N2]-[N3] is or comprises SPHSKA (SEQ ID NO: 941).
  
  177. The AAV capsid variant of any one of embodiments 168-176, which comprises an amino acid other than G at position 453 (e.g., M, T, I, E, S, A, N, V, L, K, H, P, R, W, or D), numbered according to SEQ ID NO: 138 or 981.
  
  178. The AAV capsid variant of any one of embodiments 168-177, which comprises the amino acid G at position 453, numbered according to SEQ ID NO: 138 or 981.
  
  179. The AAV capsid variant of any one of embodiments 168-178, wherein X1 of [N1] is chosen from: G, M, T, I, E, S, A, N, V, L, K, H, P, R, W, or D; optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids.
  
  180. The AAV capsid variant of any one of embodiments 168-179, wherein [N1] comprises SG, GS, MS, TS, IS, ES, SS, AS, NS, VS, LS, KS, HS, PS, RS, WS, or DS.
  
  181. The AAV capsid variant of any one of embodiments 168-180, wherein [N1] is or comprises: GSG, MSG, TSG, ISG, ESG, SSG, ASG, NSG, VSG, LSG, KSG, HSG, PSG, RSG, WSG, or DSG.
  
  182. The AAV capsid variant of any one of embodiments 168-181, wherein [N1] is or comprises GSG.
  
  183. The AAV capsid variant of any one of embodiments 168-182, wherein [N1]-[N2] comprises SGSPH (SEQ ID NO: 4752).
  
  184. The AAV capsid variant of any one of embodiments 168-183, wherein [N1]-[N2] is or comprises:

(i)

(SEQ ID NO: 4695)

GSGSPH,

(SEQ ID NO: 4798)

MSGSPH,

(SEQ ID NO: 4800)

TSGSPH,

(SEQ ID NO: 4801)

ISGSPH,

(SEQ ID NO: 4803)

ESGSPH,

(SEQ ID NO: 4804)

SSGSPH,

(SEQ ID NO: 4806)

ASGSPH,

(SEQ ID NO: 4807)

NSGSPH,

(SEQ ID NO: 4786)

VSGSPH,

(SEQ ID NO: 4808)

LSGSPH,

(SEQ ID NO: 4810)

KSGSPH,

(SEQ ID NO: 4811)

HSGSPH,

(SEQ ID NO: 4813)

PSGSPH,

(SEQ ID NO: 4815)

RSGSPH,

(SEQ ID NO: 4817)

WSGSPH,

(SEQ ID NO: 4818)

DSGSPH;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, or 5 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  185. The AAV capsid variant of any one of embodiments 168-184, wherein [N1]-[N2]-[N3] is or comprises:

(i)

(SEQ ID NO: 4697)

GSGSPHSKA,

(SEQ ID NO: 4956)

GSGSPHSKV,

(SEQ ID NO: 4957)

MSGSPHSKA,

(SEQ ID NO: 4959)

TSGSPHSKA,

(SEQ ID NO: 4960)

ISGSPHSKA,

(SEQ ID NO: 4962)

GSGSPHSKT,

(SEQ ID NO: 4963)

ESGSPHSKA,

(SEQ ID NO: 4964)

SSGSPHSKA,

(SEQ ID NO: 4953)

GSGSPHSKS,

(SEQ ID NO: 4966)

ASGSPHSKA,

(SEQ ID NO: 4967)

NSGSPHSKA,

(SEQ ID NO: 4913)

VSGSPHSKA,

(SEQ ID NO: 4968)

LSGSPHSKA,

(SEQ ID NO: 4970)

KSGSPHSKA,

(SEQ ID NO: 4972)

GSGSPHSKG,

(SEQ ID NO: 4945)

GSGSPHSKR,

(SEQ ID NO: 4973)

HSGSPHSKA,

(SEQ ID NO: 4975)

PSGSPHSKA,

(SEQ ID NO: 4977)

RSGSPHSKA,

(SEQ ID NO: 4978)

GSGSPHTKA,

(SEQ ID NO: 4980)

WSGSPHSKA,

(SEQ ID NO: 4981)

DSGSPHSKA,

(SEQ ID NO: 4983)

GSGSPHNKA,

(SEQ ID NO: 4943)

GSGSPHSKL,

(SEQ ID NO: 4994)

GSGSPHSKN,

or

(SEQ ID NO: 4995)

GSGSPHAKA;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, 6, 7, 8, or 9 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  186. The AAV capsid variant of any one of embodiments 168-185, wherein [N1]-[N2]-[N3] is or comprises GSGSPHSKA (SEQ ID NO: 4697).
  
  187. The AAV capsid variant of any one of embodiments 168-186, which comprises an amino acid other than Q at position 456 (e.g., R, P, H, L, K, I, G, S, M, or E), an amino acid other than N at position 457 (e.g., D, V, S, P, T, G, Y, W, E, R, H, K, F, A, I, L, or M), an amino acid other than Q at position 458 (e.g., R, L, A, P, H, T, I, F, K, V, M, G, W, Y, S, E, N, or D), an amino acid other than Q at position 459 (e.g., H, K, A, L, P, E, M, I, S, N, R, Y, C, V, T, W, D, G), and/or an amino acid other than T at position 460 (e.g., I, N, S, H, R, L, D, Y, A, or Q), relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  188. The AAV capsid variant of any one of embodiments 168-187, which comprises an amino acid other than Q at position 462 (e.g., R, P, H, L, K, I, G, S, M, or E), an amino acid other than N at position 463 (e.g., D, V, S, P, T, G, Y, W, E, R, H, K, F, A, I, L, or M), an amino acid other than Q at position 464 (e.g., R, L, A, P, H, T, I, F, K, V, M, G, W, Y, S, E, N, or D), an amino acid other than Q at position 465 (e.g., H, K, A, L, P, E, M, I, S, N, R, Y, C, V, T, W, D, G), and/or an amino acid other than T at position 466 (e.g., I, N, S, H, R, L, D, Y, A, or Q), relative to a reference sequence numbered according to SEQ ID NO: 981.
  
  189. The AAV capsid variant of any one of embodiments 168-188, which comprises the amino acid Q at position 456, the amino acid N at position 457, the amino acid Q at position 458, the amino acid Q at position 459, and/or the amino acid T at position 460, relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  190. The AAV capsid variant of any one of embodiments 168-189, which comprises the amino acid Q at position 462, the amino acid N at position 463, the amino acid Q at position 464, the amino acid Q at position 465, and/or the amino acid T at position 466, numbered according to SEQ ID NO: 981
  
  191. The AAV capsid variant of any one of embodiments 168-190, further comprising [N4] wherein [N4] comprises X7, X8, X9, X10, and X11, wherein:
- (a) X7 is Q, R, P, H, L, K, I, G, S, M, or E;
- (b) X8 is N, D, V, S, P, T, G, Y, W, E, R, H, K, F, A, I, L, or M;
- (c) X9 is Q, R, L, A, P, H, T, I, F, K, V, M, G, W, Y, S, E, N, D;
- (d) X10 is Q, H, K, A, L, P, E, M, I, S, N, R, Y, C, V, T, W, D, G; and
- (e) X1I is T, I, N, S, H, R, L, D, Y, A, Q;
- optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (a)-(e).
  
  192. The AAV capsid variant of embodiment 191, wherein [N4] is or comprises:

(i)

(SEQ ID NO: 5412)

QNQQT,

(SEQ ID NO: 5413)

QNRHT,

(SEQ ID NO: 5414)

RDQQT,

(SEQ ID NO: 5415)

PNLQT,

(SEQ ID NO: 5416)

HVRQT,

(SEQ ID NO: 5417)

PNQHT,

(SEQ ID NO: 5418)

QSQQT,

(SEQ ID NO: 5419)

QNQQI,

(SEQ ID NO: 5420)

QPAKT,

(SEQ ID NO: 5421)

QTQQN,

(SEQ ID NO: 5422)

QNLAT,

(SEQ ID NO: 5423)

QNQLT,

(SEQ ID NO: 5424)

QGQQT,

(SEQ ID NO: 5425)

LNRQS,

(SEQ ID NO: 5426)

HNQQT,

(SEQ ID NO: 5427)

QNPPT,

(SEQ ID NO: 5428)

QNLQT,

(SEQ ID NO: 5429)

QYQQT,

(SEQ ID NO: 5430)

QDQET,

(SEQ ID NO: 5431)

QNHQT,

(SEQ ID NO: 5432)

QDQQT,

(SEQ ID NO: 5433)

HWQQT,

(SEQ ID NO: 5434)

PNQQT,

(SEQ ID NO: 5435)

QNQLI,

(SEQ ID NO: 5436)

PEQQT,

(SEQ ID NO: 5437)

QRTMT,

(SEQ ID NO: 5438)

QNQQH,

(SEQ ID NO: 5439)

LHQHT,

(SEQ ID NO: 5440)

QHRIT,

(SEQ ID NO: 5441)

QYIHT,

(SEQ ID NO: 5442)

QKFET,

(SEQ ID NO: 5443)

QFPST,

(SEQ ID NO: 5444)

HNQQR,

(SEQ ID NO: 5445)

QAIKT,

(SEQ ID NO: 5446)

QNRQT,

(SEQ ID NO: 5447)

QYQHT,

(SEQ ID NO: 5448)

QNPQS,

(SEQ ID NO: 5449)

QHQLT,

(SEQ ID NO: 5450)

QSPPT,

(SEQ ID NO: 5451)

QAKLT,

(SEQ ID NO: 5452)

KSQQT,

(SEQ ID NO: 5453)

QDRPT,

(SEQ ID NO: 5454)

QSQQL,

(SEQ ID NO: 5455)

QAFHT,

(SEQ ID NO: 5456)

QKQQD,

(SEQ ID NO: 5457)

QNAQT,

(SEQ ID NO: 5458)

HNQLT,

(SEQ ID NO: 5459)

QNQQY,

(SEQ ID NO: 5460)

QKLNT,

(SEQ ID NO: 5461)

QNVQT,

(SEQ ID NO: 5462)

QAQQT,

(SEQ ID NO: 5463)

QNLQA,

(SEQ ID NO: 5464)

QTPPT,

(SEQ ID NO: 5465)

QYQHA,

(SEQ ID NO: 5466)

QGQQA,

(SEQ ID NO: 5467)

QPPAT,

(SEQ ID NO: 5468)

QERPT,

(SEQ ID NO: 5469)

QDLQT,

(SEQ ID NO: 5470)

QAMHT,

(SEQ ID NO: 5471)

LNQQT,

(SEQ ID NO: 5472)

QHPST,

(SEQ ID NO: 5473)

PGLQT,

(SEQ ID NO: 5474)

QGIRT,

(SEQ ID NO: 5475)

QAPAT,

(SEQ ID NO: 5476)

QSQQI,

(SEQ ID NO: 5477)

QIPPT,

(SEQ ID NO: 5478)

QTQLT,

(SEQ ID NO: 5479)

QAPST,

(SEQ ID NO: 5480)

QNTYA,

(SEQ ID NO: 5481)

QNQHI,

(SEQ ID NO: 5482)

QNHLT,

(SEQ ID NO: 5483)

QIGMT,

(SEQ ID NO: 5484)

LNKQT,

(SEQ ID NO: 5485)

QLQQT,

(SEQ ID NO: 5486)

QRMST,

(SEQ ID NO: 5487)

QGILT,

(SEQ ID NO: 5488)

QDRQT,

(SEQ ID NO: 5489)

RDWQT,

(SEQ ID NO: 5490)

QNTHD,

(SEQ ID NO: 5491)

PNLQI,

(SEQ ID NO: 5492)

QERST,

(SEQ ID NO: 5493)

QNYQT,

(SEQ ID NO: 5494)

QRTCT,

(SEQ ID NO: 5495)

QIGHT,

(SEQ ID NO: 5496)

QGAIT,

(SEQ ID NO: 5497)

QVPPT,

(SEQ ID NO: 5498)

QVQQI,

(SEQ ID NO: 5499)

LMRQT,

(SEQ ID NO: 5500)

QYSVT,

(SEQ ID NO: 5501)

QAITT,

(SEQ ID NO: 5502)

QKTLT,

(SEQ ID NO: 5503)

QNQWT,

(SEQ ID NO: 5504)

QLHHT,

(SEQ ID NO: 5505)

QNIII,

(SEQ ID NO: 5506)

QGHHT,

(SEQ ID NO: 5507)

QSKVT,

(SEQ ID NO: 5508)

QLPST,

(SEQ ID NO: 5509)

IGKQT,

(SEQ ID NO: 5510)

QAIHT,

(SEQ ID NO: 5511)

QHGLT,

(SEQ ID NO: 5512)

QFMCT,

(SEQ ID NO: 5513)

QHLQT,

(SEQ ID NO: 5514)

QNHQN,

(SEQ ID NO: 5515)

QPART,

(SEQ ID NO: 5516)

QSLQT,

(SEQ ID NO: 5517)

QSQLT,

(SEQ ID NO: 5518)

QDRQS,

(SEQ ID NO: 5519)

QMPST,

(SEQ ID NO: 5520)

QGSLT,

(SEQ ID NO: 5521)

QVPAT,

(SEQ ID NO: 5522)

QDKQT,

(SEQ ID NO: 5523)

HYQQT,

(SEQ ID NO: 5524)

QVPST,

(SEQ ID NO: 5525)

RGEQT,

(SEQ ID NO: 5526)

PGQQT,

(SEQ ID NO: 5527)

QSLQI,

(SEQ ID NO: 5528)

LEQQT,

(SEQ ID NO: 5529)

QNQST,

(SEQ ID NO: 5530)

QKVIT,

(SEQ ID NO: 5531)

QNNDQ,

(SEQ ID NO: 5532)

QSVHT,

(SEQ ID NO: 5533)

QPLGT,

(SEQ ID NO: 5534)

HNQET,

(SEQ ID NO: 5535)

QNLQI,

(SEQ ID NO: 5536)

QIQQT,

(SEQ ID NO: 5537)

QVRNT,

(SEQ ID NO: 5538)

PSNQT,

(SEQ ID NO: 5539)

QVGHT,

(SEQ ID NO: 5540)

QRDIT,

(SEQ ID NO: 5541)

QMPNT,

(SEQ ID NO: 5542)

RGLQT,

(SEQ ID NO: 5543)

QKQQT,

(SEQ ID NO: 5544)

PSLQT,

(SEQ ID NO: 5545)

QRDQT,

(SEQ ID NO: 5546)

QAKGT,

(SEQ ID NO: 5547)

QSAHT,

(SEQ ID NO: 5548)

QSTMT,

(SEQ ID NO: 5549)

QREMT,

(SEQ ID NO: 5550)

QYRAT,

(SEQ ID NO: 5551)

QWQQT,

(SEQ ID NO: 5552)

QRMNT,

(SEQ ID NO: 5553)

GDSQT,

(SEQ ID NO: 5554)

QKIST,

(SEQ ID NO: 5555)

PSMQT,

(SEQ ID NO: 5556)

SPRQT,

(SEQ ID NO: 5557)

MEQQT,

(SEQ ID NO: 5558)

QYQNT,

(SEQ ID NO: 5559)

QHQQT,

(SEQ ID NO: 5560)

INQQT,

(SEQ ID NO: 5561)

PNQQH,

(SEQ ID NO: 5562)

ENRQT,

(SEQ ID NO: 5563)

QTQQA,

or

(SEQ ID NO: 5564)

QNQAT;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, or 4 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  193. The AAV capsid variant of embodiment 191 or 192, wherein [N1]-[N2]-[N3]-[N4] is or comprises:
- (i) the amino acid sequence of any of SEQ ID NOs: 200 or 2887-3076;
- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  194. The AAV capsid variant of any one of embodiments 191-193, wherein [N1]-[N2]-[N3]-[N4] is or comprises GSGSPHSKAQNQQT (SEQ ID NO: 200).
  
  195. The AAV capsid variant of any one of embodiments 191-193, wherein [N1]-[N2]-[N3]-[N4] is or comprises VSGSPHSKAQNQQT (SEQ ID NO: 903).
  
  196. The AAV capsid variant of any one of embodiments 168-195, which comprises an amino acid other than K at position 449 (e.g., T, E, or N), T at position 450 (e.g., S, E, A, N, V, Q, or G), an amino acid other than I at position 451 (e.g., F, E, V, L, D, S, C, T, A, N, H, R, G, or W), and/or an amino acid other than N at position 452 (e.g., I, P, K, R, H, S, M, Q, D, T, L, A, Y, V, F, E, W, or G), relative to a reference sequence numbered according to SEQ ID NO: 138 or 981.
  
  197. The AAV capsid variant of any one of embodiments 168-196, which comprises the amino acid K at position 449, the amino acid T at position 450, the amino acid I at position 451, and/or the amino acid N at position 452, relative to a reference sequence numbered according to SEQ ID NO: 138 or 981.
  
  198. The AAV capsid variant of any one of embodiments 168-197, which further comprises [N0], wherein [N0] comprises X_A, X_B, X_C, and X_D, wherein:
- (a) X_Ais K, T, E, or N;
- (b) X_bis T, S, E, A, N, V, Q, or G;
- (c) X_Cis I, F, E, V, L, D, S, C, T, A, N, H, R, G, or W; and
- (d) X_Dis N, I, P, K, R, H, S, M, Q, D, T, L, A, Y, V, F, E, W, or G;
- optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (a)-(d).
  
  199. The AAV capsid variant of embodiment 198, wherein [N0] is or comprises:

(i)

(SEQ ID NO: 5565)

KTII,

(SEQ ID NO: 5566)

KTFP,

(SEQ ID NO: 5567)

KTEK,

(SEQ ID NO: 5568)

KTVN,

(SEQ ID NO: 5569)

KTFN,

(SEQ ID NO: 5570)

KTIN,

(SEQ ID NO: 5571)

TTIN,

(SEQ ID NO: 5572)

KSIN,

(SEQ ID NO: 5573)

KTER,

(SEQ ID NO: 5574)

KELH,

(SEQ ID NO: 5575)

KAIN,

(SEQ ID NO: 5576)

KTDN,

(SEQ ID NO: 5577)

KTFH,

(SEQ ID NO: 5578)

KTSN,

(SEQ ID NO: 5579)

ETIN,

(SEQ ID NO: 5580)

NTIN,

(SEQ ID NO: 5581)

KTEN,

(SEQ ID NO: 5582)

KTSS,

(SEQ ID NO: 5583)

KTCN,

(SEQ ID NO: 5584)

KTEH,

(SEQ ID NO: 5585)

KAEM,

(SEQ ID NO: 5586)

KATN,

(SEQ ID NO: 5587)

KAIK,

(SEQ ID NO: 5588)

KTDK,

(SEQ ID NO: 5589)

KTFK,

(SEQ ID NO: 5590)

KSDQ,

(SEQ ID NO: 5591)

KTEI,

(SEQ ID NO: 5592)

KTID,

(SEQ ID NO: 5593)

KNTN,

(SEQ ID NO: 5594)

KTET,

(SEQ ID NO: 5595)

KTEL,

(SEQ ID NO: 5596)

KNIN,

(SEQ ID NO: 5597)

KTEA,

(SEQ ID NO: 5598)

KTAN,

(SEQ ID NO: 5599)

NTIY,

(SEQ ID NO: 5600)

KTFS,

(SEQ ID NO: 5601)

KTES,

(SEQ ID NO: 5602)

KTTN,

(SEQ ID NO: 5603)

KTED,

(SEQ ID NO: 5604)

KTNN,

(SEQ ID NO: 5605)

KEVH,

(SEQ ID NO: 5606)

KTIS,

(SEQ ID NO: 5607)

KTVR,

(SEQ ID NO: 5608)

KTDR,

(SEQ ID NO: 5609)

ETIK,

(SEQ ID NO: 5610)

KNHI,

(SEQ ID NO: 5611)

KESD,

(SEQ ID NO: 5612)

KTIK,

(SEQ ID NO: 5613)

KTDL,

(SEQ ID NO: 5614)

KTVP,

(SEQ ID NO: 5615)

KTVI,

(SEQ ID NO: 5616)

KAEH,

(SEQ ID NO: 5617)

KNCL,

(SEQ ID NO: 5618)

KTVK,

(SEQ ID NO: 5619)

KNAD,

(SEQ ID NO: 5620)

KTIT,

(SEQ ID NO: 5621)

KNCV,

(SEQ ID NO: 5622)

KNAL,

(SEQ ID NO: 5623)

KVIN,

(SEQ ID NO: 5624)

KTEF,

(SEQ ID NO: 5625)

KTRE,

(SEQ ID NO: 5626)

KQGE,

(SEQ ID NO: 5627)

KSEK,

(SEQ ID NO: 5628)

KNVN,

(SEQ ID NO: 5629)

KGGE,

(SEQ ID NO: 5630)

KEFV,

(SEQ ID NO: 5631)

KSDK,

(SEQ ID NO: 5632)

KTEQ,

(SEQ ID NO: 5633)

KEVQ,

(SEQ ID NO: 5634)

KTEY,

(SEQ ID NO: 5635)

KNCW,

(SEQ ID NO: 5636)

KTDV,

(SEQ ID NO: 5637)

KSDI,

(SEQ ID NO: 5638)

KNSI,

(SEQ ID NO: 5639)

KNSL,

(SEQ ID NO: 5640)

KEVV,

(SEQ ID NO: 5641)

KTEP,

(SEQ ID NO: 5642)

KSEL,

(SEQ ID NO: 5643)

KTWQ,

(SEQ ID NO: 5644)

KTEV,

(SEQ ID NO: 5645)

KAVN,

(SEQ ID NO: 5646)

KGVL,

(SEQ ID NO: 5647)

KTEG,

(SEQ ID NO: 5648)

KTRD,

(SEQ ID NO: 5649)

KTGN,

(SEQ ID NO: 5650)

KNAI,

(SEQ ID NO: 5651)

KAEN,

(SEQ ID NO: 5652)

KAET,

(SEQ ID NO: 5653)

KTVH,

(SEQ ID NO: 5654)

KETA,

(SEQ ID NO: 5655)

KNNL,

(SEQ ID NO: 5656)

EAIN,

(SEQ ID NO: 5657)

KSLN,

(SEQ ID NO: 5658)

KTIP,

or

(SEQ ID NO: 5659)

KTIH;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, or 3 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  200. The AAV capsid variant of embodiment 198 or 199, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or comprises:
- (i) the amino acid sequence of any one of SEQ ID NOs: 3239-3526 or 3591-3605;
- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  201. The AAV capsid variant of any one of embodiments 198-200, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or comprises KTINGSGSPHSKAQNQQT (SEQ ID NO: 5660).
  
  202. The AAV capsid variant of any one of embodiments 198-200, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or comprises KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589).
  
  203. An AAV capsid variant comprising an amino acid sequence having the following formula: [N1]-[N2]-[N3](SEQ ID NO: 6408), wherein:
- (i) [N1] comprises positions X1, X2, and X3, wherein position X2 is an amino acid other than S and position X3 is an amino acid other than G;
- (ii) [N2] comprises the amino acid sequence SPH; and
- (iii) [N3] comprises positions X4, X5, and X6, wherein position X4 is K.
  
  204. The AAV capsid variant of embodiment 203, wherein:
- (i) X5 of [N3] is S, I, T, R, H, Y, L, or M; and
- (ii) X6 of [N3] is G, A, L, E, V, R, W, N, Q, or K;
- optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (i) or (ii).
  
  205. The AAV capsid variant of embodiment 203 or 204, wherein X5 is S and/or X₆is G.
  
  206. The AAV capsid variant of any one of embodiments 203-205, wherein [N3] comprises KS, KI, KT, KR, KH, KY, KL, KM, SG, IG, TG, RG, SA, SL, SE, SV, SR, SW, SN, HG, YG, SQ, IV, SK, LW, MG, or MA.
  
  207. The AAV capsid variant of any one of embodiments 203-206, wherein [N3] is or comprises KSG, KIG, KTG, KRG, KSA, KSL, KSE, KSV, KSR, KSW, KSN, KHG, KYG, KSQ, KIV, KSK, KLW, KMG, or KMA.
  
  208. The AAV capsid variant of any one of embodiments 203-207, wherein [N3] is or comprises KSG.
  
  209. The AAV capsid variant of any one of embodiments 203-208, wherein [N2]-[N3] comprises SPHKS (SEQ ID NO: 4704), SPHKI (SEQ ID NO: 4713), SPHKT (SEQ ID NO: 4711), SPHKR (SEQ ID NO: 4717), NPHKS (SEQ ID NO: 5661), SPHKH (SEQ ID NO: 4728), SPHKY (SEQ ID NO: 4715), SPHKL (SEQ ID NO: 4714), or SPHKM (SEQ ID NO: 4729).
  
  210. The AAV capsid variant of any one of embodiments 203-209, wherein [N2]-[N3] is or comprises:

(i)

(SEQ ID NO: 946)

SPHKSG,

(SEQ ID NO: 958)

SPHKIG,

(SEQ ID NO: 4738)

SPHKTG,

(SEQ ID NO: 974)

SPHKRG,

(SEQ ID NO: 5662)

NPHKSG,

(SEQ ID NO: 977)

SPHKSA,

(SEQ ID NO: 4740)

SPHKSL,

(SEQ ID NO: 4741)

SPHKSE,

(SEQ ID NO: 4742)

SPHKSV,

(SEQ ID NO: 951)

SPHKSR,

(SEQ ID NO: 4743)

SPHKSW,

(SEQ ID NO: 4744)

SPHKSN,

(SEQ ID NO: 4745)

SPHKHG,

(SEQ ID NO: 966)

SPHKYG,

(SEQ ID NO: 4746)

SPHKSQ,

(SEQ ID NO: 5663)

SPHKIV,

(SEQ ID NO: 4747)

SPHKSK,

(SEQ ID NO: 4748)

SPHKLW,

(SEQ ID NO: 4750)

SPHKMG,

or

(SEQ ID NO: 4751)

SPHKMA;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, or 5 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  211. The AAV capsid variant of any one of embodiments 203-210, wherein [N2]-[N3] is or comprises SPHKSG (SEQ ID NO: 946).
  
  212. The AAV capsid variant of any one of embodiments 203-211, which comprises an amino acid other than G at position 453 (e.g., A, K, W, R, L, I, M, N, T, E, Q, Y, H, F, or V), numbered according to SEQ ID NO: 138 or 981.
  
  213. The AAV capsid variant of any one of embodiments 203-212, which comprises the amino acid G at position 453, numbered according to SEQ ID NO: 138 or 981.
  
  214. The AAV capsid variant of any one of embodiments 203-214, wherein:
- (i) position X1 of [N1] is G, A, K, W, R, L, I, M, N, T, E, Q, Y, H, F, or V;
- (ii) position X2 of [N1] is H, Y, R, Q, N, P, or D;
- (iii) position X3 of [N1] is D, E, G, V, or N;
- optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (i), (ii), or (iii).
  
  215. The AAV capsid variant of any one of embodiments 203-214, wherein position X2 of [N1] is H and position X3 of [N1] is D.
  
  216. The AAV capsid variant of any one of embodiments 203-215, wherein position X1 of [N1] is G, position X2 of [N1] is H and position X3 of [N1] is D.
  
  217. The AAV capsid variant of any one of embodiments 203-216, wherein [N1] comprises GH, HD, GY, GR, GQ, AH, GN, KH, GP, WH, RH, LH, JH, MH, GD, NH, TH, EH, QH, YH, HH, FH, VH, YD, HE, RG, QD, RD, ND, PD, QV, DD, HN, or NG
  
  218. The AAV capsid variant of any one of embodiments 203-217, wherein [N1] is or comprises GHD, GYD, GHE, GRG, GQD, GRD, AHD, GND, KHD, GPD, WHD, RHD, LHD, GQV, IHD, MHD, GDD, GHN, NHD, THD, GNG, EHD, QHD, YHD, HHD, FHD, or VHD.
  
  219. The AAV capsid variant of any one of embodiments 203-218, wherein [N1] is or comprises GHD.
  
  220. The AAV capsid variant of any one of embodiments 203-219, wherein [N1]-[N2] comprises HDSPH (SEQ ID NO: 4703).
  
  221. The AAV capsid variant of any one of embodiments 203-220, wherein [N1]-[N2] is or comprises:

(i)

(SEQ ID NO: 4784)

GHDSPH,

(SEQ ID NO: 4829)

GYDSPH,

(SEQ ID NO: 4793)

GHESPH,

(SEQ ID NO: 4788)

GRGSPH,

(SEQ ID NO: 5664)

GHDNPH,

(SEQ ID NO: 4785)

GQDSPH,

(SEQ ID NO: 4831)

GRDSPH,

(SEQ ID NO: 5665)

AHDSPH,

(SEQ ID NO: 4832)

GNDSPH,

(SEQ ID NO: 5666)

KHDSPH,

(SEQ ID NO: 4833)

GPDSPH,

(SEQ ID NO: 5667)

WHDSPH,

(SEQ ID NO: 5668)

RHDSPH,

(SEQ ID NO: 5669)

LHDSPH,

(SEQ ID NO: 4835)

GQVSPH,

(SEQ ID NO: 5670)

IHDSPH,

(SEQ ID NO: 5671)

MHDSPH,

(SEQ ID NO: 4792)

GDDSPH,

(SEQ ID NO: 4836)

GHNSPH,

(SEQ ID NO: 5672)

NHDSPH,

(SEQ ID NO: 5673)

THDSPH,

(SEQ ID NO: 4805)

GNGSPH,

(SEQ ID NO: 5674)

EHDSPH,

(SEQ ID NO: 5675)

QHDSPH,

(SEQ ID NO: 5676)

YHDSPH,

(SEQ ID NO: 5677)

HHDSPH,

(SEQ ID NO: 5678)

FHDSPH,

or

(SEQ ID NO: 5679)

VHDSPH;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, or 5 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  222. The AAV capsid variant of any one of embodiments 203-221, wherein [N1]-[N2]-[N3] is or comprises:

(i)

(SEQ ID NO: 4698)

GHDSPHKSG,

(SEQ ID NO: 4996)

GHDSPHKIG,

(SEQ ID NO: 4997)

GYDSPHKSG,

(SEQ ID NO: 4998)

GHESPHKSG,

(SEQ ID NO: 4999)

GHDSPHKTG,

(SEQ ID NO: 5000)

GRGSPHKRG,

(SEQ ID NO: 5680)

GHDNPHKSG,

(SEQ ID NO: 4908)

GQDSPHKSG,

(SEQ ID NO: 4940)

GHDSPHKSA,

(SEQ ID NO: 5001)

GHDSPHKSL,

(SEQ ID NO: 5003)

GHDSPHKSE,

(SEQ ID NO: 5004)

GRDSPHKSG,

(SEQ ID NO: 5681)

AHDSPHKSG,

(SEQ ID NO: 5005)

GNDSPHKSV,

(SEQ ID NO: 5682)

AHDSPHKIG,

(SEQ ID NO: 4939)

GHESPHKSA,

(SEQ ID NO: 5006)

GQDSPHKIG,

(SEQ ID NO: 5007)

GHDSPHKSV,

(SEQ ID NO: 4942)

GHDSPHKSR,

(SEQ ID NO: 5683)

KHDSPHKSG,

(SEQ ID NO: 5008)

GPDSPHKIG,

(SEQ ID NO: 5009)

GPDSPHKSG,

(SEQ ID NO: 5010)

GHDSPHKSW,

(SEQ ID NO: 5684)

WHDSPHKSG,

(SEQ ID NO: 5685)

RHDSPHKSG,

(SEQ ID NO: 5011)

GHDSPHKSN,

(SEQ ID NO: 4937)

GHDSPHKRG,

(SEQ ID NO: 5013)

GHDSPHKHG,

(SEQ ID NO: 5686)

LHDSPHKSG,

(SEQ ID NO: 5014)

GQVSPHKSG,

(SEQ ID NO: 5687)

IHDSPHKSG,

(SEQ ID NO: 5688)

MHDSPHKSG,

(SEQ ID NO: 5015)

GDDSPHKSV,

(SEQ ID NO: 5016)

GHNSPHKSG,

(SEQ ID NO: 5689)

NHDSPHKSG,

(SEQ ID NO: 5690)

THDSPHKSG,

(SEQ ID NO: 5017)

GNGSPHKRG,

(SEQ ID NO: 5691)

EHDSPHKSG,

(SEQ ID NO: 5018)

GHDSPHKYG,

(SEQ ID NO: 5019)

GHDSPHKSQ,

(SEQ ID NO: 5692)

QHDSPHKSG,

(SEQ ID NO: 5693)

RHDSPHKIV,

(SEQ ID NO: 5694)

YHDSPHKSG,

(SEQ ID NO: 5020)

GNDSPHKIG,

(SEQ ID NO: 5695)

HHDSPHKSG,

(SEQ ID NO: 5021)

GHDSPHKSK,

(SEQ ID NO: 5696)

FHDSPHKSG,

(SEQ ID NO: 5022)

GHDSPHKLW,

(SEQ ID NO: 5697)

VHDSPHKSG,

(SEQ ID NO: 5024)

GHDSPHKMG,

(SEQ ID NO: 5025)

GHDSPHKMA,

or

(SEQ ID NO: 4938)

GDDSPHKSG;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, 6, 7, 8, or 9 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  223. The AAV capsid variant of any one of embodiments 203-222, wherein [N1]-[N2]-[N3] is or comprises GHDSPHKSG (SEQ ID NO: 4698).
  
  224. The AAV capsid variant of any one of embodiments 203-223, which comprises an amino acid other than Q at position 456 (e.g., R, P, H, K, L, V, A, E, or I), an amino acid other than N at position
  
  457 (e.g., I, K, S, H, R, T, D, Y, L, W, F, A, Q, or M), an amino acid other than Q at position 458 (e.g., R, V, K, P, Y, H, L, I, E, or M), an amino acid other than Q at position 459 (e.g., H, L, E, P, W, D, I, V, S, K, R, C, M, or N), and/or an amino acid other than T at position 460 (e.g., A, E, K, S, I, P, G, or N), relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  225. The AAV capsid variant of any one of embodiments 203-224, which comprises an amino acid other than Q at position 462 (e.g., R, P, H, K, L, V, A, E, or I), an amino acid other than N at position 463 (e.g., I, K, S, H, R, T, D, Y, L, W, F, A, Q, or M), an amino acid other than Q at position 464 (e.g., R, V, K, P, Y, H, L, I, E, or M), an amino acid other than Q at position 465 (e.g., H, L, E, P, W, D, I, V, S, K, R, C, M, or N), and/or an amino acid other than T at position 466 (e.g., A, E, K, S, I, P, G, or N), relative to a reference sequence numbered according to SEQ ID NO: 982.
  
  226. The AAV capsid variant of any one of embodiments 203-225, which comprises the amino acid Q at position 456, the amino acid N at position 457, the amino acid Q at position 458, the amino acid Q at position 459, and/or the amino acid T at position 460, relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  227. The AAV capsid variant of any one of embodiments 203-226, which comprises the amino acid Q at position 462, the amino acid N at position 463, the amino acid Q at position 464, the amino acid Q at position 465, and/or the amino acid T at position 466, relative to a reference sequence numbered according to SEQ ID NO: 138.
  
  228. The AAV capsid variant of any one of embodiments 203-227, further comprising [N4] wherein [N4] comprises X7, X8, X9, X10, and X11, wherein:
- (a) X7 is Q, R, P, H, L, K, I, G, S, M, or E;
- (b) X8 is N, D, V, S, P, T, G, Y, W, E, R, H, K, F, A, I, L, or M;
- (c) X9 is Q, R, L, A, P, H, T, I, F, K, V, M, G, W, Y, S, E, N, D;
- (d) X10 is Q, H, K, A, L, P, E, M, I, S, N, R, Y, C, V, T, W, D, G; and
- (e) X11 is T, I, N, S, H, R, L, D, Y, A, Q;
- optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (a)-(e).
  
  229. The AAV capsid variant of embodiment 228, wherein [N4] is or comprises:

(i)

(SEQ ID NO: 5412)

QNQQT,

(SEQ ID NO: 5698)

QIRQT,

(SEQ ID NO: 5699)

QNQHA,

(SEQ ID NO: 5543)

QKQQT,

(SEQ ID NO: 5700)

QSVQT,

(SEQ ID NO: 5701)

RSQQT,

(SEQ ID NO: 5702)

QNKLE,

(SEQ ID NO: 5703)

QNQQK,

(SEQ ID NO: 5704)

QHQQA,

(SEQ ID NO: 5705)

QIQHT,

(SEQ ID NO: 5706)

PRQQT,

(SEQ ID NO: 5707)

HTQQT,

(SEQ ID NO: 5708)

QRQHT,

(SEQ ID NO: 5418)

QSQQT,

(SEQ ID NO: 5709)

QNQQS,

(SEQ ID NO: 5710)

RNQET,

(SEQ ID NO: 5478)

QTQLT,

(SEQ ID NO: 5711)

KNQQT,

(SEQ ID NO: 5432)

QDQQT,

(SEQ ID NO: 5426)

HNQQT,

(SEQ ID NO: 5423)

QNQLT,

(SEQ ID NO: 5712)

QTQQT,

(SEQ ID NO: 5713)

QTQQI,

(SEQ ID NO: 5714)

QSKQA,

(SEQ ID NO: 5715)

QNQPP,

(SEQ ID NO: 5716)

QSPQT,

(SEQ ID NO: 5493)

QNYQT,

(SEQ ID NO: 5431)

QNHQT,

(SEQ ID NO: 5446)

QNRQT,

(SEQ ID NO: 5717)

QNQQG,

(SEQ ID NO: 5482)

QNHLT,

(SEQ ID NO: 5447)

QYQHT,

(SEQ ID NO: 5503)

QNQWT,

(SEQ ID NO: 5718)

QNQHT,

(SEQ ID NO: 5719)

QTRQT,

(SEQ ID NO: 5720)

QNLHT,

(SEQ ID NO: 5471)

LNQQT,

(SEQ ID NO: 5721)

QNQET,

(SEQ ID NO: 5513)

QHLQT,

(SEQ ID NO: 5722)

LNQPT,

(SEQ ID NO: 5723)

QNQDT,

(SEQ ID NO: 5724)

RNQQT,

(SEQ ID NO: 5725)

QNLLT,

(SEQ ID NO: 5726)

QLVIT,

(SEQ ID NO: 5727)

RTQET,

(SEQ ID NO: 5728)

QTHQT,

(SEQ ID NO: 5729)

QNQPA,

(SEQ ID NO: 5730)

QDQHT,

(SEQ ID NO: 5731)

QSQHT,

(SEQ ID NO: 5732)

RNQQI,

(SEQ ID NO: 5733)

VRQQT,

(SEQ ID NO: 5734)

QNQHS,

(SEQ ID NO: 5735)

AWQQT,

(SEQ ID NO: 5736)

QSVPT,

(SEQ ID NO: 5737)

QNIQP,

(SEQ ID NO: 5738)

QNHLN,

(SEQ ID NO: 5739)

LDQQT,

(SEQ ID NO: 5740)

PDQQS,

(SEQ ID NO: 5741)

ESQQT,

(SEQ ID NO: 5742)

QNKQT,

(SEQ ID NO: 5743)

QRQLT,

(SEQ ID NO: 5744)

QIIVT,

(SEQ ID NO: 5745)

QKQST,

(SEQ ID NO: 5746)

QSHQT,

(SEQ ID NO: 5747)

QFVVT,

(SEQ ID NO: 5428)

QNLQT,

(SEQ ID NO: 5419)

QNQQI,

(SEQ ID NO: 5748)

QSQPT,

(SEQ ID NO: 5749)

QNEQT,

(SEQ ID NO: 5516)

QSLQT,

(SEQ ID NO: 5750)

RNRQT,

(SEQ ID NO: 5751)

QSKQT,

(SEQ ID NO: 5752)

QNPLT,

(SEQ ID NO: 5753)

RDQKT,

(SEQ ID NO: 5754)

HNQQN,

(SEQ ID NO: 5755)

QWKRT,

(SEQ ID NO: 5476)

QSQQI,

(SEQ ID NO: 5462)

QAQQT,

(SEQ ID NO: 5756)

QNHQI,

(SEQ ID NO: 5757)

QNQQA,

(SEQ ID NO: 5758)

QNQLN,

(SEQ ID NO: 5759)

QTQPT,

(SEQ ID NO: 5560)

INQQT,

(SEQ ID NO: 5760)

QKQLT,

(SEQ ID NO: 5761)

RNQLA,

(SEQ ID NO: 5762)

RNQQS,

(SEQ ID NO: 5763)

ISIQT,

(SEQ ID NO: 5764)

QNQQN,

(SEQ ID NO: 5765)

QSQQS,

(SEQ ID NO: 5766)

QTVCT,

(SEQ ID NO: 5767)

QYQQI,

(SEQ ID NO: 5768)

QQIMT,

(SEQ ID NO: 5769)

QNEQS,

(SEQ ID NO: 5770)

LNHQT,

(SEQ ID NO: 5771)

QMIHT,

(SEQ ID NO: 5772)

RNHQS,

(SEQ ID NO: 5773)

QKMNT,

(SEQ ID NO: 5774)

QSQQN,

(SEQ ID NO: 5465)

QYQHA;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, or 4 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  230. The AAV capsid variant of embodiment 228 or 229, wherein [N1]-[N2]-[N3]-[N4] is or comprises:
- (i) the amino acid sequence of any of SEQ ID NOs: 201 or 3160-3237;
- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  231. The AAV capsid variant of any one of embodiments 228-230, wherein [N1]-[N2]-[N3]-[N4] is or comprises GHDSPHKSGQNQQT (SEQ ID NO: 201).
  
  232. The AAV capsid variant of any one of embodiments 203-231, which comprises an amino acid other than K at position 449 (e.g., T), T at position 450 (e.g., A, S, I, V, N, E, Y, C, G, W, or Q), an amino acid other than I at position 451 (e.g., E, V, S, T, N, D, C, G, Q, L, P, A), and/or an amino acid other than N at position 452 (e.g., S, Y, I, K, F, T, D, E, G, V, L, A, M, Q, H, P, or R), relative to a reference sequence numbered according to SEQ ID NO: 138 or 982.
  
  233. The AAV capsid variant of any one of embodiments 203-232, which comprises the amino acid K at position 449, the amino acid T at position 450, the amino acid I at position 451, and/or the amino acid N at position 452, relative to a reference sequence numbered according to SEQ ID NO: 138 or 982.
  
  234. The AAV capsid variant of any one of embodiments 203-233, which further comprises [N0], wherein [N0] comprises X_A, X_B, X_C, and X_D, wherein:
- (a) X_Ais K or T;
- (b) X_bis T, A, S, I, V, N, E, Y, C, G, W, or Q;
- (c) X_Cis I, E, V, S, T, N, D, C, G, Q, L, P, A; and
- (d) X_Dis N, S, Y, I, K, F, T, D, E, G, V, L, A, M, Q, H, P, or R;
- optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (a)-(d).
  
  235. The AAV capsid variant of embodiment 234, wherein [N0] is or comprises:

(i)

(SEQ ID NO: 5575)

KAIN,

(SEQ ID NO: 5570)

KTIN,

(SEQ ID NO: 5601)

KTES,

(SEQ ID NO: 5571)

TTIN,

(SEQ ID NO: 5572)

KSIN,

(SEQ ID NO: 5568)

KTVN,

(SEQ ID NO: 5775)

KSIY,

(SEQ ID NO: 5578)

KTSN,

(SEQ ID NO: 5602)

KTTN,

(SEQ ID NO: 5776)

KIIN,

(SEQ ID NO: 5606)

KTIS,

(SEQ ID NO: 5777)

KAII,

(SEQ ID NO: 5612)

KTIK,

(SEQ ID NO: 5624)

KTEF,

(SEQ ID NO: 5620)

KTIT,

(SEQ ID NO: 5604)

KTNN,

(SEQ ID NO: 5592)

KTID,

(SEQ ID NO: 5778)

KAIS,

(SEQ ID NO: 5779)

KTVD,

(SEQ ID NO: 5780)

KTIE,

(SEQ ID NO: 5647)

KTEG,

(SEQ ID NO: 5623)

KVIN,

(SEQ ID NO: 5645)

KAVN,

(SEQ ID NO: 5781)

KTIY,

(SEQ ID NO: 5576)

KTDN,

(SEQ ID NO: 5583)

KTCN,

(SEQ ID NO: 5782)

KNVV,

(SEQ ID NO: 5595)

KTEL,

(SEQ ID NO: 5783)

KTDA,

(SEQ ID NO: 5644)

KTEV,

(SEQ ID NO: 5642)

KSEL,

(SEQ ID NO: 5784)

KTEM,

(SEQ ID NO: 5632)

KTEQ,

(SEQ ID NO: 5565)

KTII,

(SEQ ID NO: 5785)

KIVN,

(SEQ ID NO: 5567)

KTEK,

(SEQ ID NO: 5581)

KTEN,

(SEQ ID NO: 5786)

KIGN,

(SEQ ID NO: 5787)

KEVM,

(SEQ ID NO: 5788)

KYQV,

(SEQ ID NO: 5597)

KTEA,

(SEQ ID NO: 5586)

KATN,

(SEQ ID NO: 5584)

KTEH,

(SEQ ID NO: 5789)

KTVE,

(SEQ ID NO: 5790)

KAID,

(SEQ ID NO: 5791)

KTIM,

(SEQ ID NO: 5792)

KEVG,

(SEQ ID NO: 5793)

KSEM,

(SEQ ID NO: 5794)

KAQQ,

(SEQ ID NO: 5795)

KCGE,

(SEQ ID NO: 5796)

KASN,

(SEQ ID NO: 5594)

KTET,

(SEQ ID NO: 5797)

KTIG,

(SEQ ID NO: 5798)

KTDP,

(SEQ ID NO: 5799)

KELV,

(SEQ ID NO: 5800)

KELM,

(SEQ ID NO: 5801)

KNEI,

(SEQ ID NO: 5802)

KTPN,

(SEQ ID NO: 5803)

KITN,

(SEQ ID NO: 5804)

KTDI,

(SEQ ID NO: 5805)

KTDQ,

(SEQ ID NO: 5806)

KGIN,

(SEQ ID NO: 5807)

KSEI,

(SEQ ID NO: 5627)

KSEK,

(SEQ ID NO: 5808)

KWSA,

(SEQ ID NO: 5809)

KELA,

(SEQ ID NO: 5810)

KQTQ,

(SEQ ID NO: 5811)

KGAD,

(SEQ ID NO: 5812)

KVGE,

(SEQ ID NO: 5813)

KANE,

(SEQ ID NO: 5814)

KTDT,

(SEQ ID NO: 5815)

KTCI,

(SEQ ID NO: 5816)

KELR,

(SEQ ID NO: 5817)

KCQI,

(SEQ ID NO: 5818)

KGVM,

(SEQ ID NO: 5819)

KACD,

(SEQ ID NO: 5820)

KNEL,

(SEQ ID NO: 5821)

KAAE,

(SEQ ID NO: 5822)

KGQN,

(SEQ ID NO: 5823)

KNEF,

(SEQ ID NO: 5824)

KTSI,

(SEQ ID NO: 5616)

KAEH,

(SEQ ID NO: 5825)

KCDQ,

(SEQ ID NO: 5826)

KEIL,

(SEQ ID NO: 5573)

KTER,

(SEQ ID NO: 5650)

KNAI,

(SEQ ID NO: 5588)

KTDK,

(SEQ ID NO: 5827)

KTPD,

(SEQ ID NO: 5659)

KTIH,

or

(SEQ ID NO: 5591)

KTEI;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, or 3 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  236. The AAV capsid variant of embodiment 234 or 235, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or comprises:
- (i) the amino acid sequence of any one of SEQ ID NOs: 3606-3836;
- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  237. The AAV capsid variant of any one of embodiments 234-236, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or comprises KTINGHDSPHKSGQNQQT (SEQ ID NO: 5828).
  
  238. The AAV capsid variant of any one of embodiments 234-236, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or comprises KAEIGHDSPHKSGQNQQT (SEQ ID NO: 1754).
  
  239. The AAV capsid variant of any one of embodiments 234-236, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or comprises KTEKMSGSPHSKAQNQQT (SEQ ID NO: 3241).
  
  240. The AAV capsid variant of any one of embodiments 168-239, wherein [N1]-[N2]-[N3] is present in loop IV.
  
  241. The AAV capsid variant of any one of embodiments 198-202 or 234-240, wherein [N0] and [N4] are present in loop IV.
  
  242. The AAV capsid variant of any one of embodiments 198-202 or 234-241, wherein [N0] is present immediately subsequent to position 448, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138, 981, or 982.
  
  243. The AAV capsid variant of any one of embodiments 198-202 or 234-242, wherein [N0] replaces positions 449-452 (e.g., K449, T450, I451, and N452), relative to a reference sequence numbered according to SEQ ID NO: 138, 981, or 982.
  
  244. The AAV capsid variant of any one of embodiments 198-202 or 234-243, wherein [N0] is present immediately subsequent to position 448 and wherein [N0] replaces positions 449-452 (e.g., K449, T450, 1451, and N452), relative to a reference sequence numbered according to SEQ ID NO: 138, 981, or 982.
  
  245. The AAV capsid variant of any one of embodiments 198-202 or 234-244, wherein [N0] corresponds to positions 449-452 (e.g., K449, T450, 1451, and N452) of any one of SEQ ID NOs: 138, 981, or 982.
  
  246. The AAV capsid variant of any one of embodiments 168-245, wherein [N1] is present immediately subsequent to position 452, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138, 981, or 982.
  
  247. The AAV capsid variant of any one of embodiments 168-246, wherein [N1] replaces positions 453-455 (e.g., G453, S454, and G455), relative to a reference sequence numbered according to SEQ ID NO: 138, 981, or 982.
  
  248. The AAV capsid variant of any one of embodiments 168-246, wherein [N1] replaces position 453 (e.g., G453), relative to a reference sequence numbered according to SEQ ID NO: 138, 981, or 982.
  
  249. The AAV capsid variant of any one of embodiments 168-177, 179-181, 183-185, 187-193, 195-200, 202, or 240-246, wherein:
- (i) position X1 of [N1] replaces position 453 (e.g., G453);
- (ii) position X2 of [N1] corresponds to position 454 (e.g., S454); and
- (iii) position X3 of [N1] corresponds to position 455 (e.g., G455),
- wherein (i), (ii), and (iii) are numbered according to SEQ ID NO: 138 or SEQ ID NO: 981.
  
  250. The AAV capsid variant of any one of embodiments 168-176, 178-201, or 240-246, wherein:
- (i) position X1 of [N1] corresponds to position 453 (e.g., G453);
- (ii) position X2 of [N1] corresponds to position 454 (e.g., S454); and
- (iii) position X3 of [N1] corresponds to position 455 (e.g., G455);
- wherein (i), (ii), and (iii) are numbered according to SEQ ID NO: 138 or SEQ ID NO: 981.
  
  251. The AAV capsid variant of any one of embodiments 203-248, wherein:
- (i) position X1 of [N1] corresponds to position 453 (e.g., G453);
- (ii) position X2 of [N1] replaces position 454 (e.g., S454); and
- (iii) position X3 of [N1] replaces position 455 (e.g., G455),
- wherein (i), (ii), and (iii) are numbered according to SEQ ID NO: 138 or SEQ ID NO: 982.
  
  252. The AAV capsid variant of any one of embodiments 203-248 or 251, wherein [N1] corresponds to positions 453-455 (e.g., G453, H454, D455) of SEQ ID NO 982.
  
  253. The AAV capsid variant of any one of embodiments 168-176, 178-201, 240-247, or 250, wherein [N1] corresponds to positions 453-455 (e.g., G453, S454, G455) of SEQ ID NO: 138, or 981.
  
  254. The AAV capsid variant of any one of embodiments 168-253, wherein [N2] is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138, 981, or 982.
  
  255. The AAV capsid variant of any one of embodiments 168-254, wherein [N2] corresponds to positions 456-458 (e.g., S456, P457, and H458) of SEQ ID NO: 981 or 982.
  
  256. The AAV capsid variant of any one of embodiments 168-254, wherein [N2] corresponds to positions 456-458 (e.g., S456, P457, and H458) of any one of SEQ ID NOs: 36-59.
  
  257. The AAV capsid variant of any one of embodiments 168-256, wherein [N2] is present immediately subsequent to [N1].
  
  258. The AAV capsid variant of any one of embodiments 168-202, 240-247, 249-257, wherein [N3] corresponds to positions 459-460 (e.g., S459, K460, A461) of SEQ ID NO: 981.
  
  259. The AAV capsid variant of any one of embodiments 168-202, 240-247, 249-257, wherein [N3] corresponds to positions 459-460 (e.g., S459, K460, A461) of SEQ ID NO: 36, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 57, or 59.
  
  260. The AAV capsid variant of any one of embodiments 168-259, wherein [N2]-[N3] is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of any one of SEQ ID NOs: 138, 981, or 982.
  
  261. The AAV capsid variant of any one of embodiments 168-259, wherein [N2]-[N3] is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981.
  
  262. The AAV capsid variant of any one of embodiments 168-261, wherein [N2]-[N3] corresponds to positions 456-461 (e.g., S456, P457, H458, S459, K460, A461) of SEQ ID NO: 981.
  
  263. The AAV capsid variant of any one of embodiments 168-262, wherein [N2]-[N3] corresponds to positions 456-461 (e.g., S456, P457, H458, S459, K460, A461) of any one of SEQ ID NOs: 36, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 57, or 59.
  
  264. The AAV capsid variant of any one of embodiments 203-257 or 259-261, wherein [N3] corresponds to positions 459-460 (e.g., K459, S460, G461) of SEQ ID NO: 982.
  
  265. The AAV capsid variant of any one of embodiments 203-257 or 259-261, wherein [N3] corresponds to positions 459-460 (e.g., K459, S460, G461) of SEQ ID NO: 37.
  
  266. The AAV capsid variant of any one of embodiments 203-265, wherein [N2]-[N3] is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 982.
  
  267. The AAV capsid variant of any one of embodiments 203-246, 248, 252, 255, 257, 260, 263-266, wherein [N3] replaces positions 454 and 455 (e.g., S454 and G455), numbered according to SEQ ID NO: 138.
  
  268. The AAV capsid variant of any one of embodiments 203-246, 248, 252, 255, 257, 260, 263-267, wherein [N3] is present immediately subsequent to [N2] and replaces positions 454 and 455 (e.g., S454 and G455), numbered according to SEQ ID NO: 138.
  
  269. The AAV capsid variant of any one of embodiments 203-246, 248, 252, 255, 257, 260, 263-268, wherein [N3] is present immediately subsequent to [N1]-[N2] and replaces positions 454 and 455 (e.g., S454 and G455), numbered according to SEQ ID NO: 138.
  
  270. The AAV capsid variant of any one of embodiments 203-257, 260, 264-269, wherein [N2]-[N3] corresponds to positions 456-461 (e.g., S456, P457, H458, K459, S460, G461) of SEQ ID NO: 982.
  
  271. The AAV capsid variant of any one of embodiments 203-257, 260, 264-270, wherein [N2]-[N3] corresponds to positions 456-461 (e.g., S456, P457, H458, K459, S460, G461) of SEQ ID NO: 37.
  
  272. The AAV capsid variant of any one of embodiments 191-202 or 228-271, wherein [N4] is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  273. The AAV capsid variant of any one of embodiments 191-202 or 228-272, wherein [N4] replaces positions 456-460 (e.g., Q456, N457, Q458, Q459, and T460), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  274. The AAV capsid variant of any one of embodiments 191-202 or 228-273, wherein [N4] corresponds to positions 462-466 (e.g., Q462, N463, Q464, Q465, and T466) of SEQ ID NO: 981 or 982.
  
  275. The AAV capsid variant of any one of embodiments 191-202 or 228-273, wherein [N4] corresponds to positions 462-466 of any one of SEQ ID NOs: 36-59.
  
  276. The AAV capsid variant of any one of embodiments 191-202 or 228-274, wherein [N4] corresponds to positions 456-460 (e.g., Q456, N457, Q458, Q459, and T460) of SEQ ID NO: 138.
  
  277. The AAV capsid variant of any one of embodiments 191-202 or 228-276, wherein [N2]-[N3]-[N4] replaces positions 456-460 (e.g., Q456, N457, Q458, Q459, and T460), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  278. The AAV capsid variant of any one of embodiments 191-202 or 228-277, wherein [N2]-[N3]-[N4] is present immediately subsequent to position 455, and wherein [N2]-[N3]-[N4] replaces positions 456-460 (e.g., Q456, N457, Q458, Q459, and T460), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  279. The AAV capsid variant of any one of embodiments 191-202 or 228-278, wherein [N1]-[N2]-[N3]-[N4] replaces positions 453-460 (e.g., G453, S454, G455, Q456, N457, Q458, Q459, and T460), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  280. The AAV capsid variant of any one of embodiments 191-202 or 228-279, wherein [N1]-[N2]-[N3]-[N4] is present immediately subsequent to position 452, and wherein [N1]-[N2]-[N3]-[N4] replaces positions 453-460 (e.g., G453, S454, G455, Q456, N457, Q458, Q459, and T460), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  281. The AAV capsid variant of any one of embodiments 191-202, 240-247, 249, 250, 253-263, 266, or 272-280, wherein [N1]-[N2]-[N3]-[N4] corresponds to positions 453-466 (e.g., G453, S454, G455, S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465, and T466) of SEQ ID NO: 981.
  
  282. The AAV capsid variant of any one of embodiments 168-202, 240-247, 249, 250, 253-263, 266, or 272-280, wherein [N1]-[N2]-[N3] corresponds to positions 453-461 (e.g., G453, S454, G455, S456, P457, H458, S459, K460, A461) of SEQ ID NO: 981.
  
  283. The AAV capsid variant of any one of embodiments 228-257, 260, 261, 264-282, wherein [N1]-[N2]-[N3]-[N4] corresponds to positions 453-466 (e.g., G453, H454, D455, S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465, T466) of SEQ ID NO: 982.
  
  284. The AAV capsid variant of any one of embodiments 203-257, 260, 261, 264-283, wherein [N1]-[N2]-[N3] corresponds to positions 453-461 (e.g., G453, H454, D455, S456, P457, H458, K459, S460, G461) of SEQ ID NO: 982.
  
  285. The AAV capsid variant of any one of embodiments 228-257, 260, 261, 264-282, wherein [N1]-[N2]-[N3]-[N4] corresponds to positions 453-466 of any one of SEQ ID NOs: 36-59.
  
  286. The AAV capsid variant of any one of embodiments 198-202 or 234-286, wherein [N0]-[N1]-[N2]-[N3]-[N4] replaces positions 449-460 (e.g., K449, T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, Q459, and T460), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  287. The AAV capsid variant of any one of embodiments 198-202 or 234-286, wherein [N0]-[N1]-[N2]-[N3]-[N4] is present immediately subsequent to position 449, and wherein [N0]-[N1]-[N2]-[N3]-[N4] replaces positions 449-460 (e.g., K449, T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, Q459, and T460), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  288. The AAV capsid variant of any one of embodiments 198-202, 240-247, 249, 250, 253-263, 266, 272-281, 286, or 287, wherein [N0]-[N1]-[N2]-[N3]-[N4] corresponds to positions 449-466 (e.g., K449, T450, 1451, N452, G453, S454, G455, S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465, T466) of SEQ ID NO: 981.
  
  289. The AAV capsid variant of any one of embodiments 234-257, 260, 261, 264-284, 286, or 287, wherein [N0]-[N1]-[N2]-[N3]-[N4] corresponds to positions 449-466 (e.g., K449, T450, 1451, N452, G453, H454, D455, S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465, T466) of SEQ ID NO: 982.
  
  290. The AAV capsid variant of any one of embodiments 234-257, 260, 261, 264-284, 286, or 287, wherein [N0]-[N1]-[N2]-[N3]-[N4] corresponds to positions 449-466 of any one of SEQ ID NOs: 36-59.
  
  291. The AAV capsid variant of any one of embodiments 191-202 or 228-290, wherein [N4] is present immediately subsequent to position 461, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981 or 982.
  
  292. The AAV capsid variant of any one of embodiments 191-202 or 228-291, wherein [N4] replaces positions 462-466 (e.g., Q462, N463, Q464, Q465, and T466), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981 or 982.
  
  293. The AAV capsid variant of any one of embodiments 191-202 or 228-292, wherein [N2]-[N3]-[N4] replaces positions 462-466 (e.g., Q462, N463, Q464, Q465, and T466), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981 or 982.
  
  294. The AAV capsid variant of any one of embodiments 191-202 or 228-293, wherein [N2]-[N3]-[N4] is present immediately subsequent to position 455, and wherein [N2]-[N3]-[N4] replaces positions 462-466 (e.g., Q462, N463, Q464, Q465, and T466), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981 or 982.
  
  295. The AAV capsid variant of any one of embodiments 168-294, which comprises from N-terminus to C-terminus, [N2]-[N3].
  
  296. The AAV capsid variant of any one of embodiments 168-295, which comprises from N-terminus to C-terminus, [N1]-[N2]-[N3].
  
  297. The AAV capsid variant of any one of embodiments 168-296, which comprises from N-terminus to C-terminus, [N0]-[N1]-[N2]-[N3].
  
  298. The AAV capsid variant of any one of embodiments 168-297, which comprises from N-terminus to C-terminus, [N1]-[N2]-[N3]-[N4].
  
  299. The AAV capsid variant of any one of embodiments 168-298, which comprises from N-terminus to C-terminus, [N0]-[N1]-[N2]-[N3]-[N4].
  
  300. An AAV capsid variant comprising the formula [A]-[B](SEQ ID NO: 4696), wherein:
- (i) [A] comprises GSGSPH (SEQ ID NO: 4695); and
- (ii) [B] comprises X1 X2, X3, X4, and X5, wherein:
  - (a) position X1 is S, I, F, V, C, Y, W, R, P, L, Q, M, K, or G;
  - (b) position X2 is K, M, R, F, V, C, P, Y, L, W, G, N, S, T, I, or A;
  - (c) position X3 is A, Y, L, R, W, C, T, F, H, I, P, M, K, S, V, G, Q, or N;
  - (d) position X4 is Q, M, F, K, H, R, C, W, P, V, L, G, S, Y, I, A, T, D, N, or E; and
  - (e) position X5 is A, N, Y, R, K, L, I, M, Q, S, C, W, F, T, G, V, or P; optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (a)-(e).
    
    301. The AAV capsid variant of embodiment 300, wherein:
- (a) position X1 is S, L, R, V, or P;
- (b) position X2 is K, C, F, L, P, R, S, or V;
- (c) position X3 is A, C, F, I, K, L, M, P, R, T, W, or Y;
- (d) position X4 is Q, R, S, T, C, F, K, L, P or Y; and
- (e) position X5 is N, R, S, T, K, M, Q or Y;
- optionally wherein the AAV capsid variant comprises an amino acid modification, e.g., a conservative substitution, of any of the aforesaid amino acids in (a)-(e).
  
  302. The AAV capsid variant of embodiment 300 or 301, wherein [B] comprises SKA, SMY, SKL, SKR, SKW, SRC, SFT, SKF, IVW, SKY, SCH, FPW, SKI, VYY, SLY, SKP, SRF, SRM, SVK, SWA, SLW, SFR, SKK, SYA, SCS, SGA, SFP, SFF, SMC, SKT, SGK, FYR, CRV, YGI, VNC, SLA, WSY, RWL, PSC, SSW, SKG, VPW, SGC, STT, PKR, SKC, WVP, SFW, RIK, SKM, LRW, LPT, SYM, LLC, RCC, LCV, SYL, QGC, MAF, SFQ, SLC, RPW, RPR, SCP, SVR, SLP, VYH, SYT, LVY, YRY, SWL, CPA, SPP, RWT, PRK, PFV, SKS, WVA, SKV, CAL, SSC, SKN, LCT, STC, SKQ, KSG, SYY, SLT, SCQ, FPF, SVF, GRY, AQA, AQN, YMN, AFY, LKR, RHR, AQK, WRL, CRN, TCN, FFI, AQY, WQN, YFM, ARQ, HQN, IRR, YQN, YWN, AFS, FWN, AQC, MRN, KKN, APN, WKN, ARW, RPN, KVF, AFN, ACS, RLW, SRN, CPN, ACN, FRQ, PFN, FGN, CQN, LFW, TRK, KRN, RQN, VQN, IQN, AQR, PFR, AWN, RSY, LQN, WLN, RRA, AQT, GCT, RYT, TPN, ARM, CFL, PQN, WSN, FKN, KQN, APR, RYN, MIC, TQN, WKS, AAR, LTR, IRG, LVN, FQN, ACQ, WGL, ILR, QIN, ACI, ALR, AHA, CLN, AFV, AQF, RCN, MPC, KTS, PYN, AQS, TRN, LKN, AQM, CTN, PDN, RNY, ACR, CSV, ARI, LPK, SEQ, VRM, NSR, RKR, ARN, QRP, RVV, GQN, YSN, QSN, AKG, CTS, FEN, AKK, KAQ, MYM, KAF, KLK, KRH, KWR, RCR, FTC, KFF, VWQ, KYF, KAR, CHQ, PWQ, KIR, YYQ, LYW, KPK, RFW, RMR, VKK, WAP, LWK, FRP, KKV, YAF, KAC, KRL, CSR, RCP, GAC, KFR, FFG, MCQ, KLF, KTR, GKR, YRQ, RVQ, GIQ, NCQ, KPF, LAW, KRS, SYQ, WLQ, KRR, KGC, KRY, GCQ, FTP, TTC, KRQ, KCF, VPQ, FWS, KFK, IKQ, KAP, FRY, KMI, RWQ, PTQ, KWK, YMR, KAA, LCQ, CCQ, CVQ, KLT, KLC, YLV, AFQ, KWG, KIL, FQI, KAL, KAH, LCL, PRQ, CPQ, VRY, VRC, KMP, KKT, LPY, YHQ, YTR, VYQ, RYQ, WLK, PAQ, MCT, PPD, WTQ, RKQ, KCS, FVQ, KLP, KSE, VAQ, LYQ, KVR, ALQ, SCT, KNS, KRK, CTQ, TCL, YAR, KQR, KRV, SGQ, YYS, LTC, CQS, KAK, KPQ, PFQ, KCT, or VFE.
  
  303. The AAV capsid variant of any one of embodiments 300-302, wherein [B] comprises SKAQ (SEQ ID NO: 5829), SMYM (SEQ ID NO: 5830), SKAF (SEQ ID NO: 5831), SKLK (SEQ ID NO: 5832), SKRH (SEQ ID NO: 5833), SKWR (SEQ ID NO: 5834), SRCR (SEQ ID NO: 5835), SFTC (SEQ ID NO: 5836), SKFF (SEQ ID NO: 5837), IVWQ (SEQ ID NO: 5838), SKYF (SEQ ID NO: 5839), SKAR (SEQ ID NO: 5840), SCHQ (SEQ ID NO: 5841), FPWQ (SEQ ID NO: 5842), SKIR (SEQ ID NO: 5843), VYYQ (SEQ ID NO: 5844), SLYW (SEQ ID NO: 5845), SKPK (SEQ ID NO: 5846), SRFW (SEQ ID NO: 5847), SRMR (SEQ ID NO: 5848), SVKK (SEQ ID NO: 5849), SWAP (SEQ ID NO: 5850), SLWK (SEQ ID NO: 5851), SFRP (SEQ ID NO: 5852), SKKV (SEQ ID NO: 5853), SYAF (SEQ ID NO: 5854), SKAC (SEQ ID NO: 5855), SKRL (SEQ ID NO: 5856), SCSR (SEQ ID NO: 5857), SRCP (SEQ ID NO: 5858), SGAC (SEQ ID NO: 5859), SKFR (SEQ ID NO: 5860), SFPF (SEQ ID NO: 5861), SFFG (SEQ ID NO: 5862), SMCQ (SEQ ID NO: 5863), SKLF (SEQ ID NO: 5864), SKTR (SEQ ID NO: 5865), SGKR (SEQ ID NO: 5866), FYRQ (SEQ ID NO: 5867), CRVQ (SEQ ID NO: 5868), YGIQ (SEQ ID NO: 5869), VNCQ (SEQ ID NO: 5870), SKPF (SEQ ID NO: 5871), SLAW (SEQ ID NO: 5872), SKRS (SEQ ID NO: 5873), WSYQ (SEQ ID NO: 5874), RWLQ (SEQ ID NO: 5875), PSCQ (SEQ ID NO: 5876), SSWL (SEQ ID NO: 5877), SKRR (SEQ ID NO: 5878), SKGC (SEQ ID NO: 5879), VPWQ (SEQ ID NO: 5880), SKRY (SEQ ID NO: 5881), SGCQ (SEQ ID NO: 5882), SFTP (SEQ ID NO: 5883), STTC (SEQ ID NO: 5884), PKRQ (SEQ ID NO: 5885), SKCF (SEQ ID NO: 5886), WVPQ (SEQ ID NO: 5887), SFWS (SEQ ID NO: 5888), SKFK (SEQ ID NO: 5889), RIKQ (SEQ ID NO: 5890), SKAP (SEQ ID NO: 5891), SFRY (SEQ ID NO: 5892), SKMI (SEQ ID NO: 5893), LRWQ (SEQ ID NO: 5894), LPTQ (SEQ ID NO: 5895), SKWK (SEQ ID NO: 5896), SYMR (SEQ ID NO: 5897), SKAA (SEQ ID NO: 5898), LLCQ (SEQ ID NO: 5899), RCCQ (SEQ ID NO: 5900), LCVQ (SEQ ID NO: 5901), SKLT (SEQ ID NO: 5902), SKLC (SEQ ID NO: 5903), SYLV (SEQ ID NO: 5904), QGCQ (SEQ ID NO: 5905), MAFQ (SEQ ID NO: 5906), SKWG (SEQ ID NO: 5907), SKIL (SEQ ID NO: 5908), SFQI (SEQ ID NO: 5909), SKAL (SEQ ID NO: 5910), SKAH (SEQ ID NO: 5911), SLCL (SEQ ID NO: 5912), RPWQ (SEQ ID NO: 5913), RPRQ (SEQ ID NO: 5914), SCPQ (SEQ ID NO: 5915), SVRY (SEQ ID NO: 5916), SVRC (SEQ ID NO: 5917), SKMP (SEQ ID NO: 5918), SKKT (SEQ ID NO: 5919), SLPY (SEQ ID NO: 5920), VYHQ (SEQ ID NO: 5921), SYTR (SEQ ID NO: 5922), LVYQ (SEQ ID NO: 5923), YRYQ (SEQ ID NO: 5924), SWLK (SEQ ID NO: 5925), CPAQ (SEQ ID NO: 5926), SMCT (SEQ ID NO: 5927), SPPD (SEQ ID NO: 5928), SKRN (SEQ ID NO: 5929), RWTQ (SEQ ID NO: 5930), PRKQ (SEQ ID NO: 5931), SKCS (SEQ ID NO: 5932), PFVQ (SEQ ID NO: 5933), SKLP (SEQ ID NO: 5934), SKSE (SEQ ID NO: 5935), WVAQ (SEQ ID NO: 5936), SLYQ (SEQ ID NO: 5937), SKVR (SEQ ID NO: 5938), CALQ (SEQ ID NO: 5939), SSCT (SEQ ID NO: 5940), SKNS (SEQ ID NO: 5941), SKRK (SEQ ID NO: 5942), LCTQ (SEQ ID NO: 5943), STCL (SEQ ID NO: 5944), SYAR (SEQ ID NO: 5945), SKQR (SEQ ID NO: 5946), SKRV (SEQ ID NO: 5947), KSGQ (SEQ ID NO: 5948), SYYS (SEQ ID NO: 5949), SLTC (SEQ ID NO: 5950), SCQS (SEQ ID NO: 5951), SKAK (SEQ ID NO: 5952), SKPQ (SEQ ID NO: 5953), FPFQ (SEQ ID NO: 5954), SKCT (SEQ ID NO: 5955), SVFE (SEQ ID NO: 5956), GRYQ (SEQ ID NO: 5957), KAQA (SEQ ID NO: 5958), KAQN (SEQ ID NO: 5959), MYMN (SEQ ID NO: 5960), KAFY (SEQ ID NO: 5961), KLKR (SEQ ID NO: 5962), KRHR (SEQ ID NO: 5963), KAQK (SEQ ID NO: 5964), KWRL (SEQ ID NO: 5965), RCRN (SEQ ID NO: 5966), FTCN (SEQ ID NO: 5967), KFFI (SEQ ID NO: 5968), KAQY (SEQ ID NO: 5969), VWQN (SEQ ID NO: 5970), KYFM (SEQ ID NO: 5971), KARQ (SEQ ID NO: 5972), CHQN (SEQ ID NO: 5973), PWQN (SEQ ID NO: 5974), KIRR (SEQ ID NO: 5975), YYQN (SEQ ID NO: 5976), LYWN (SEQ ID NO: 5977), KPKR (SEQ ID NO: 5978), KAFS (SEQ ID NO: 5979), RFWN (SEQ ID NO: 5980), KAQC (SEQ ID NO: 5981), RMRN (SEQ ID NO: 5982), VKKN (SEQ ID NO: 5983), WAPN (SEQ ID NO: 5984), LWKN (SEQ ID NO: 5985), KARW (SEQ ID NO: 5986), FRPN (SEQ ID NO: 5987), KKVF (SEQ ID NO: 5988), YAFN (SEQ ID NO: 5989), KACS (SEQ ID NO: 5990), KRLW (SEQ ID NO: 5991), CSRN (SEQ ID NO: 5992), RCPN (SEQ ID NO: 5993), GACN (SEQ ID NO: 5994), KFRQ (SEQ ID NO: 5995), FPFN (SEQ ID NO: 5996), FFGN (SEQ ID NO: 5997), MCQN (SEQ ID NO: 5998), KLFW (SEQ ID NO: 5999), KTRK (SEQ ID NO: 6000), GKRN (SEQ ID NO: 6001), YRQN (SEQ ID NO: 6002), RVQN (SEQ ID NO: 6003), GIQN (SEQ ID NO: 6004), KAQR (SEQ ID NO: 6005), NCQN (SEQ ID NO: 6006), KPFR (SEQ ID NO: 6007), LAWN (SEQ ID NO: 6008), KRSY (SEQ ID NO: 6009), SYQN (SEQ ID NO: 6010), WLQN (SEQ ID NO: 6011), SCQN (SEQ ID NO: 6012), SWLN (SEQ ID NO: 6013), KRRA (SEQ ID NO: 6014), KAQT (SEQ ID NO: 6015), KGCT (SEQ ID NO: 6016), KRYT (SEQ ID NO: 6017), GCQN (SEQ ID NO: 6018), FTPN (SEQ ID NO: 6019), TTCN (SEQ ID NO: 6020), KARM (SEQ ID NO: 6021), KRQN (SEQ ID NO: 6022), KCFL (SEQ ID NO: 6023), VPQN (SEQ ID NO: 6024), FWSN (SEQ ID NO: 6025), KFKN (SEQ ID NO: 6026), IKQN (SEQ ID NO: 6027), KAPR (SEQ ID NO: 6028), FRYN (SEQ ID NO: 6029), KMIC (SEQ ID NO: 6030), RWQN (SEQ ID NO: 6031), PTQN (SEQ ID NO: 6032), KWKS (SEQ ID NO: 6033), YMRN (SEQ ID NO: 6034), KAAR (SEQ ID NO: 6035), LCQN (SEQ ID NO: 6036), CCQN (SEQ ID NO: 6037), CVQN (SEQ ID NO: 6038), KLTR (SEQ ID NO: 6039), KLCT (SEQ ID NO: 6040), KIRG (SEQ ID NO: 6041), YLVN (SEQ ID NO: 6042), AFQN (SEQ ID NO: 6043), KACQ (SEQ ID NO: 6044), KWGL (SEQ ID NO: 6045), KILR (SEQ ID NO: 6046), FQIN (SEQ ID NO: 6047), KACI (SEQ ID NO: 6048), KALR (SEQ ID NO: 6049), KAHA (SEQ ID NO: 6050), LCLN (SEQ ID NO: 6051), KAFV (SEQ ID NO: 6052), PRQN (SEQ ID NO: 6053), CPQN (SEQ ID NO: 6054), KAQF (SEQ ID NO: 6055), VRYN (SEQ ID NO: 6056), VRCN (SEQ ID NO: 6057), KMPC (SEQ ID NO: 6058), KKTS (SEQ ID NO: 6059), LPYN (SEQ ID NO: 6060), YHQN (SEQ ID NO: 6061), KAQS (SEQ ID NO: 6062), YTRN (SEQ ID NO: 6063), VYQN (SEQ ID NO: 6064), RYQN (SEQ ID NO: 6065), WLKN (SEQ ID NO: 6066), KAQM (SEQ ID NO: 6067), PAQN (SEQ ID NO: 6068), MCTN (SEQ ID NO: 6069), PPDN (SEQ ID NO: 6070), KRNY (SEQ ID NO: 6071), WTQN (SEQ ID NO: 6072), KACR (SEQ ID NO: 6073), RKQN (SEQ ID NO: 6074), KCSV (SEQ ID NO: 6075), KARI (SEQ ID NO: 6076), FVQN (SEQ ID NO: 6077), KLPK (SEQ ID NO: 6078), KSEQ (SEQ ID NO: 6079), VAQN (SEQ ID NO: 6080), LYQN (SEQ ID NO: 6081), KVRM (SEQ ID NO: 6082), ALQN (SEQ ID NO: 6083), SCTN (SEQ ID NO: 6084), KNSR (SEQ ID NO: 6085), KRKR (SEQ ID NO: 6086), CTQN (SEQ ID NO: 6087), TCLN (SEQ ID NO: 6088), YARN (SEQ ID NO: 6089), KQRP (SEQ ID NO: 6090), KRVV (SEQ ID NO: 6091), SGQN (SEQ ID NO: 6092), YYSN (SEQ ID NO: 6093), LTCN (SEQ ID NO: 6094), CQSN (SEQ ID NO: 6095), KAKG (SEQ ID NO: 6096), KPQN (SEQ ID NO: 6097), PFQN (SEQ ID NO: 6098), KCTS (SEQ ID NO: 6099), VFEN (SEQ ID NO: 6100), or KAKK (SEQ ID NO: 6101).
  
  304. The AAV capsid variant of any one of embodiments 300-303, wherein [B] is or comprises:

(i)

(SEQ ID NO: 6102)

SKAQA,

(SEQ ID NO: 6103)

SKAQN,

(SEQ ID NO: 6104)

SMYMN,

(SEQ ID NO: 6105)

SKAFY,

(SEQ ID NO: 6106)

SKLKR,

(SEQ ID NO: 6107)

SKRHR,

(SEQ ID NO: 6108)

SKAQK,

(SEQ ID NO: 6109)

SKWRL,

(SEQ ID NO: 6110)

SRCRN,

(SEQ ID NO: 6111)

SFTCN,

(SEQ ID NO: 6112)

SKFFI,

(SEQ ID NO: 6113)

SKAQY,

(SEQ ID NO: 6114)

IVWQN,

(SEQ ID NO: 6115)

SKYFM,

(SEQ ID NO: 6116)

SKARQ,

(SEQ ID NO: 6117)

SCHQN,

(SEQ ID NO: 6118)

FPWQN,

(SEQ ID NO: 6119)

SKIRR,

(SEQ ID NO: 6120)

VYYQN,

(SEQ ID NO: 6121)

SLYWN,

(SEQ ID NO: 6122)

SKPKR,

(SEQ ID NO: 6123)

SKAFS,

(SEQ ID NO: 6124)

SRFWN,

(SEQ ID NO: 6125)

SKAQC,

(SEQ ID NO: 6126)

SRMRN,

(SEQ ID NO: 6127)

SVKKN,

(SEQ ID NO: 6128)

SWAPN,

(SEQ ID NO: 6129)

SLWKN,

(SEQ ID NO: 6130)

SKARW,

(SEQ ID NO: 6131)

SFRPN,

(SEQ ID NO: 6132)

SKKVF,

(SEQ ID NO: 6133)

SYAFN,

(SEQ ID NO: 6134)

SKACS,

(SEQ ID NO: 6135)

SKRLW,

(SEQ ID NO: 6136)

SCSRN,

(SEQ ID NO: 6137)

SRCPN,

(SEQ ID NO: 6138)

SGACN,

(SEQ ID NO: 6139)

SKFRQ,

(SEQ ID NO: 6140)

SFPFN,

(SEQ ID NO: 6141)

SFFGN,

(SEQ ID NO: 6142)

SMCQN,

(SEQ ID NO: 6143)

SKLFW,

(SEQ ID NO: 6144)

SKTRK,

(SEQ ID NO: 6145)

SGKRN,

(SEQ ID NO: 6146)

FYRQN,

(SEQ ID NO: 6147)

CRVQN,

(SEQ ID NO: 6148)

YGIQN,

(SEQ ID NO: 6149)

SKAQR,

(SEQ ID NO: 6150)

VNCQN,

(SEQ ID NO: 6151)

SKPFR,

(SEQ ID NO: 6152)

SLAWN,

(SEQ ID NO: 6153)

SKRSY,

(SEQ ID NO: 6154)

WSYQN,

(SEQ ID NO: 6155)

RWLQN,

(SEQ ID NO: 6156)

PSCQN,

(SEQ ID NO: 6157)

SSWLN,

(SEQ ID NO: 6158)

SKRRA,

(SEQ ID NO: 6159)

SKAQT,

(SEQ ID NO: 6160)

SKGCT,

(SEQ ID NO: 6161)

VPWQN,

(SEQ ID NO: 6162)

SKRYT,

(SEQ ID NO: 6163)

SGCQN,

(SEQ ID NO: 6164)

SFTPN,

(SEQ ID NO: 6165)

STTCN,

(SEQ ID NO: 6166)

SKARM,

(SEQ ID NO: 6167)

PKRQN,

(SEQ ID NO: 6168)

SKCFL,

(SEQ ID NO: 6169)

WVPQN,

(SEQ ID NO: 6170)

SFWSN,

(SEQ ID NO: 6171)

SKFKN,

(SEQ ID NO: 6172)

RIKQN,

(SEQ ID NO: 6173)

SKAPR,

(SEQ ID NO: 6174)

SFRYN,

(SEQ ID NO: 6175)

SKMIC,

(SEQ ID NO: 6176)

LRWQN,

(SEQ ID NO: 6177)

LPTQN,

(SEQ ID NO: 6178)

SKWKS,

(SEQ ID NO: 6179)

SYMRN,

(SEQ ID NO: 6180)

SKAAR,

(SEQ ID NO: 6181)

LLCQN,

(SEQ ID NO: 6182)

RCCQN,

(SEQ ID NO: 6183)

LCVQN,

(SEQ ID NO: 6184)

SKLTR,

(SEQ ID NO: 6185)

SKLCT,

(SEQ ID NO: 6186)

SKIRG,

(SEQ ID NO: 6187)

SYLVN,

(SEQ ID NO: 6188)

QGCQN,

(SEQ ID NO: 6189)

MAFQN,

(SEQ ID NO: 6190)

SKACQ,

(SEQ ID NO: 6191)

SKWGL,

(SEQ ID NO: 6192)

SKILR,

(SEQ ID NO: 6193)

SFQIN,

(SEQ ID NO: 6194)

SKACI,

(SEQ ID NO: 6195)

SKALR,

(SEQ ID NO: 6196)

SKAHA,

(SEQ ID NO: 6197)

SLCLN,

(SEQ ID NO: 6198)

SKAFV,

(SEQ ID NO: 6199)

RPWQN,

(SEQ ID NO: 6200)

RPRQN,

(SEQ ID NO: 6201)

SCPQN,

(SEQ ID NO: 6202)

SKAQF,

(SEQ ID NO: 6203)

SVRYN,

(SEQ ID NO: 6204)

SVRCN,

(SEQ ID NO: 6205)

SKMPC,

(SEQ ID NO: 6206)

SKKTS,

(SEQ ID NO: 6207)

SLPYN,

(SEQ ID NO: 6208)

VYHQN,

(SEQ ID NO: 6209)

SKAQS,

(SEQ ID NO: 6210)

SYTRN,

(SEQ ID NO: 6211)

LVYQN,

(SEQ ID NO: 6212)

YRYQN,

(SEQ ID NO: 6213)

SWLKN,

(SEQ ID NO: 6214)

SKAQM,

(SEQ ID NO: 6215)

CPAQN,

(SEQ ID NO: 6216)

SMCTN,

(SEQ ID NO: 6217)

SPPDN,

(SEQ ID NO: 6218)

SKRNY,

(SEQ ID NO: 6219)

RWTQN,

(SEQ ID NO: 6220)

SKACR,

(SEQ ID NO: 6221)

PRKQN,

(SEQ ID NO: 6222)

SKCSV,

(SEQ ID NO: 6223)

SKARI,

(SEQ ID NO: 6224)

PFVQN,

(SEQ ID NO: 6225)

SKLPK,

(SEQ ID NO: 6226)

SKSEQ,

(SEQ ID NO: 6227)

WVAQN,

(SEQ ID NO: 6228)

SLYQN,

(SEQ ID NO: 6229)

SKVRM,

(SEQ ID NO: 6230)

CALQN,

(SEQ ID NO: 6231)

SSCTN,

(SEQ ID NO: 6232)

SKNSR,

(SEQ ID NO: 6233)

SKRKR,

(SEQ ID NO: 6234)

LCTQN,

(SEQ ID NO: 6235)

STCLN,

(SEQ ID NO: 6236)

SYARN,

(SEQ ID NO: 6237)

SKQRP,

(SEQ ID NO: 6238)

SKRVV,

(SEQ ID NO: 6239)

KSGQN,

(SEQ ID NO: 6240)

SYYSN,

(SEQ ID NO: 6241)

SLTCN,

(SEQ ID NO: 6242)

SCQSN,

(SEQ ID NO: 6243)

SKAKG,

(SEQ ID NO: 6244)

SKPQN,

(SEQ ID NO: 6245)

FPFQN,

(SEQ ID NO: 6246)

SKCTS,

(SEQ ID NO: 6247)

SVFEN,

(SEQ ID NO: 6248)

SKAKK,

or

(SEQ ID NO: 6249)

GRYQN;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, or 4 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  305. The AAV capsid variant of any one of embodiments 300-304, wherein [A]-[B] is or comprises:

(i)

(SEQ ID NO: 6250)

GSGSPHSKAQA,

(SEQ ID NO: 6251)

GSGSPHSKAQN,

(SEQ ID NO: 6252)

GSGSPHSMYMN,

(SEQ ID NO: 6253)

GSGSPHSKAFY,

(SEQ ID NO: 6254)

GSGSPHSKLKR,

(SEQ ID NO: 6255)

GSGSPHSKRHR,

(SEQ ID NO: 6256)

GSGSPHSKAQK,

(SEQ ID NO: 6257)

GSGSPHSKWRL,

(SEQ ID NO: 6258)

GSGSPHSRCRN,

(SEQ ID NO: 6259)

GSGSPHSFTCN,

(SEQ ID NO: 6260)

GSGSPHSKFFI,

(SEQ ID NO: 6261)

GSGSPHSKAQY,

(SEQ ID NO: 6262)

GSGSPHIVWQN,

(SEQ ID NO: 6263)

GSGSPHSKYFM,

(SEQ ID NO: 6264)

GSGSPHSKARQ,

(SEQ ID NO: 6265)

GSGSPHSCHON,

(SEQ ID NO: 6266)

GSGSPHFPWQN,

(SEQ ID NO: 6267)

GSGSPHSKIRR,

(SEQ ID NO: 6268)

GSGSPHVYYQN,

(SEQ ID NO: 6269)

GSGSPHSLYWN,

(SEQ ID NO: 6270)

GSGSPHSKPKR,

(SEQ ID NO: 6271)

GSGSPHSKAFS,

(SEQ ID NO: 6272)

GSGSPHSRFWN,

(SEQ ID NO: 6273)

GSGSPHSKAQC,

(SEQ ID NO: 6274)

GSGSPHSRMRN,

(SEQ ID NO: 6275)

GSGSPHSVKKN,

(SEQ ID NO: 6276)

GSGSPHSWAPN,

(SEQ ID NO: 6277)

GSGSPHSLWKN,

(SEQ ID NO: 6278)

GSGSPHSKARW,

(SEQ ID NO: 6279)

GSGSPHSFRPN,

(SEQ ID NO: 6280)

GSGSPHSKKVF,

(SEQ ID NO: 6281)

GSGSPHSYAFN,

(SEQ ID NO: 6282)

GSGSPHSKACS,

(SEQ ID NO: 6283)

GSGSPHSKRLW,

(SEQ ID NO: 6284)

GSGSPHSCSRN,

(SEQ ID NO: 6285)

GSGSPHSRCPN,

(SEQ ID NO: 6286)

GSGSPHSGACN,

(SEQ ID NO: 6287)

GSGSPHSKFRQ,

(SEQ ID NO: 6288)

GSGSPHSFPFN,

(SEQ ID NO: 6289)

GSGSPHSFFGN,

(SEQ ID NO: 6290)

GSGSPHSMCQN,

(SEQ ID NO: 6291)

GSGSPHSKLFW,

(SEQ ID NO: 6292)

GSGSPHSKTRK,

(SEQ ID NO: 6293)

GSGSPHSGKRN,

(SEQ ID NO: 6294)

GSGSPHFYRQN,

(SEQ ID NO: 6295)

GSGSPHCRVQN,

(SEQ ID NO: 6296)

GSGSPHYGIQN,

(SEQ ID NO: 6297)

GSGSPHSKAQR,

(SEQ ID NO: 6298)

GSGSPHVNCQN,

(SEQ ID NO: 6299)

GSGSPHSKPFR,

(SEQ ID NO: 6300)

GSGSPHSLAWN,

(SEQ ID NO: 6301)

GSGSPHSKRSY,

(SEQ ID NO: 6302)

GSGSPHWSYQN,

(SEQ ID NO: 6303)

GSGSPHRWLQN,

(SEQ ID NO: 6304)

GSGSPHPSCQN,

(SEQ ID NO: 6305)

GSGSPHSSWLN,

(SEQ ID NO: 6306)

GSGSPHSKRRA,

(SEQ ID NO: 6307)

GSGSPHSKAQT,

(SEQ ID NO: 6308)

GSGSPHSKGCT,

(SEQ ID NO: 6309)

GSGSPHVPWQN,

(SEQ ID NO: 6310)

GSGSPHSKRYT,

(SEQ ID NO: 6311)

GSGSPHSGCQN,

(SEQ ID NO: 6312)

GSGSPHSFTPN,

(SEQ ID NO: 6313)

GSGSPHSTTCN,

(SEQ ID NO: 6314)

GSGSPHSKARM,

(SEQ ID NO: 6315)

GSGSPHPKRQN,

(SEQ ID NO: 6316)

GSGSPHSKCFL,

(SEQ ID NO: 6317)

GSGSPHWVPQN,

(SEQ ID NO: 6318)

GSGSPHSFWSN,

(SEQ ID NO: 6319)

GSGSPHSKFKN,

(SEQ ID NO: 6320)

GSGSPHRIKQN,

(SEQ ID NO: 6321)

GSGSPHSKAPR,

(SEQ ID NO: 6322)

GSGSPHSFRYN,

(SEQ ID NO: 6323)

GSGSPHSKMIC,

(SEQ ID NO: 6324)

GSGSPHLRWQN,

(SEQ ID NO: 6325)

GSGSPHLPTQN,

(SEQ ID NO: 6326)

GSGSPHSKWKS,

(SEQ ID NO: 6327)

GSGSPHSYMRN,

(SEQ ID NO: 6328)

GSGSPHSKAAR,

(SEQ ID NO: 6329)

GSGSPHLLCQN,

(SEQ ID NO: 6330)

GSGSPHRCCQN,

(SEQ ID NO: 6331)

GSGSPHLCVQN,

(SEQ ID NO: 6332)

GSGSPHSKLTR,

(SEQ ID NO: 6333)

GSGSPHSKLCT,

(SEQ ID NO: 6334)

GSGSPHSKIRG,

(SEQ ID NO: 6335)

GSGSPHSYLVN,

(SEQ ID NO: 6336)

GSGSPHQGCON,

(SEQ ID NO: 6337)

GSGSPHMAFQN,

(SEQ ID NO: 6338)

GSGSPHSKACQ,

(SEQ ID NO: 6339)

GSGSPHSKWGL,

(SEQ ID NO: 6340)

GSGSPHSKILR,

(SEQ ID NO: 6341)

GSGSPHSFQIN,

(SEQ ID NO: 6342)

GSGSPHSKACI,

(SEQ ID NO: 6343)

GSGSPHSKALR,

(SEQ ID NO: 6344)

GSGSPHSKAHA,

(SEQ ID NO: 6345)

GSGSPHSLCLN,

(SEQ ID NO: 6346)

GSGSPHSKAFV,

(SEQ ID NO: 6347)

GSGSPHRPWQN,

(SEQ ID NO: 6348)

GSGSPHRPRQN,

(SEQ ID NO: 6349)

GSGSPHSCPQN,

(SEQ ID NO: 6350)

GSGSPHSKAQF,

(SEQ ID NO: 6351)

GSGSPHSVRYN,

(SEQ ID NO: 6352)

GSGSPHSVRCN,

(SEQ ID NO: 6353)

GSGSPHSKMPC,

(SEQ ID NO: 6354)

GSGSPHSKKTS,

(SEQ ID NO: 6355)

GSGSPHSLPYN,

(SEQ ID NO: 6356)

GSGSPHVYHQN,

(SEQ ID NO: 6357)

GSGSPHSKAQS,

(SEQ ID NO: 6358)

GSGSPHSYTRN,

(SEQ ID NO: 6359)

GSGSPHLVYQN,

(SEQ ID NO: 6360)

GSGSPHYRYQN,

(SEQ ID NO: 6361)

GSGSPHSWLKN,

(SEQ ID NO: 6362)

GSGSPHSKAQM,

(SEQ ID NO: 6363)

GSGSPHCPAQN,

(SEQ ID NO: 6364)

GSGSPHSMCTN,

(SEQ ID NO: 6365)

GSGSPHSPPDN,

(SEQ ID NO: 6366)

GSGSPHSKRNY,

(SEQ ID NO: 6367)

GSGSPHRWTQN,

(SEQ ID NO: 6368)

GSGSPHSKACR,

(SEQ ID NO: 6369)

GSGSPHPRKQN,

(SEQ ID NO: 6370)

GSGSPHSKCSV,

(SEQ ID NO: 6371)

GSGSPHSKARI,

(SEQ ID NO: 6372)

GSGSPHPFVQN,

(SEQ ID NO: 6373)

GSGSPHSKLPK,

(SEQ ID NO: 6374)

GSGSPHSKSEQ,

(SEQ ID NO: 6375)

GSGSPHWVAQN,

(SEQ ID NO: 6376)

GSGSPHSLYQN,

(SEQ ID NO: 6377)

GSGSPHSKVRM,

(SEQ ID NO: 6378)

GSGSPHCALQN,

(SEQ ID NO: 6379)

GSGSPHSSCTN,

(SEQ ID NO: 6380)

GSGSPHSKNSR,

(SEQ ID NO: 6381)

GSGSPHSKRKR,

(SEQ ID NO: 6382)

GSGSPHLCTQN,

(SEQ ID NO: 6383)

GSGSPHSTCLN,

(SEQ ID NO: 6384)

GSGSPHSYARN,

(SEQ ID NO: 6385)

GSGSPHSKQRP,

(SEQ ID NO: 6386)

GSGSPHSKRVV,

(SEQ ID NO: 6387)

GSGSPHKSGQN,

(SEQ ID NO: 6388)

GSGSPHSYYSN,

(SEQ ID NO: 6389)

GSGSPHSLTCN,

(SEQ ID NO: 6390)

GSGSPHSCQSN,

(SEQ ID NO: 6391)

GSGSPHSKAKG,

(SEQ ID NO: 6392)

GSGSPHSKPQN,

(SEQ ID NO: 6393)

GSGSPHFPFQN,

(SEQ ID NO: 6394)

GSGSPHSKCTS,

(SEQ ID NO: 6395)

GSGSPHSVFEN,

(SEQ ID NO: 6396)

GSGSPHSKAKK,

or

(SEQ ID NO: 6397)

GSGSPHGRYQN;

- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i)
  
  306. The AAV capsid of any one of embodiments 300-305, wherein [A]-[B] does not comprise the amino acid sequence of GSGSPHSKAQN (SEQ ID NO: 6251).
  
  307. The AAV capsid variant of any one of embodiments 300-306, which comprises one, two, or all of an amino acid other than Q at position 458 (e.g., R, C, S, W, L, F, Y, H, I, V, A, or P), an amino acid other than Q at position 459 (e.g., K, I, R, L or S), and/or an amino acid other than T at position 460 (e.g., R), numbered relative to SEQ ID NO: 138.
  
  308. The AAV capsid variant of any one of embodiments 300-307, which comprises:
- (i) the amino acid R at position 458;
- (ii) the amino acid W at position 458;
- (iii) the amino acid Y at position 458;
- (iv) the amino acid F at position 458;
- (v) the amino acid S at position 458;
- (vi) the amino acid C at position 458;
- (vii) the amino acid I at position 458;
- (viii) the amino acid L at position 458;
- (ix) the amino acid P at position 458;
- (x) the amino acid I at position 459;
- (xi) the amino acid H at position 458; or
- (xii) the amino acid V at position 458;
- wherein (i)-(xii) are numbered according to SEQ ID NO: 138.
  
  309. The AAV capsid variant of any one of embodiments 300-307, which comprises:
- (i) the amino acid R at position 458 and the amino acid K at position 459;
- (ii) the amino acid C at position 458 and the amino acid I at position 459;
- (iii) the amino acid S at position 458 and the amino acid R at position 459′
- (iv) the amino acid L at position 458 and the amino acid K at position 459;
- (v) the amino acid F at position 458 and the amino acid K at position 459;
- (vi) the amino acid C at position 458 and the amino acid R at position 459;
- (vii) the amino acid H at position 458 and the amino acid R at position 459;
- (viii) the amino acid I at position 458 and the amino acid L at position 459;
- (ix) the amino acid V at position 458 and the amino acid R at position 459;
- (x) the amino acid A at position 458 and the amino acid K at position 459;
- (xi) the amino acid I at position 458 and the amino acid K at position 459;
- (xii) the amino acid C at position 458 and the amino acid S at position 459; or
- (xiii) the amino acid C at position 458 and the amino acid L at position 459
- wherein (i)-(xiii) are numbered according to SEQ ID NO: 138.
  
  310. The AAV capsid variant of any one of embodiments 300-307, which comprises the amino acid F at position 458, the amino acid K at position 459, and the amino acid R at position 460, numbered relative to SEQ ID NO: 138.
  
  311. The AAV capsid variant of any one of embodiments 300-310, which comprises one, two, or all of an amino acid other than T at position 450 (e.g., Y, P, W, R, K, S, or F), an amino acid other than I at position 451 (e.g., R, S, Y, L, V, H, P, A, or F), and/or an amino acid other than N at position 452 (e.g., V, W, A, T, F, Y, L, R, H, S, or M), numbered relative to SEQ ID NO: 138.
  
  312. The AAV capsid variant of any one of embodiments 300-311, which comprises the amino acid V at position 452, numbered according to SEQ ID NO: 138.
  
  313. The AAV capsid variant of any one of embodiments 300-312, which comprises the amino acid Y at position 450 and the amino acid V at position 452, numbered according to SEQ ID NO: 138.
  
  314. The AAV capsid variant of any one of embodiments 300-312, which comprises the amino acid R at position 450 and the amino acid Y at position 451, numbered according to SEQ ID NO: 138.
  
  315. The AAV capsid variant of any one of embodiments 300-311, which comprises:
- (i) the amino acid P at position 450, the amino acid R at position 451, and the amino acid W at position 452;
- (ii) the amino acid Y at position 450, the amino acid S at position 451, and the amino acid A at position 452;
- (iii) the amino acid Y at position 450, the amino acid Y at position 451, and the amino acid T at position 452;
- (iv) the amino acid P at position 450, the amino acid R at position 451, and the amino acid F at position 452;
- (v) the amino acid W at position 450, the amino acid L at position 451, and the amino acid T at position 452;
- (vi) the amino acid R at position 450, the amino acid S at position 451, and the amino acid Y at position 452;
- (vii) the amino acid Y at position 450, the amino acid V at position 451, and the amino acid F at position 452;
- (viii) the amino acid K at position 450, the amino acid H at position 451, and the amino acid L at position 452;
- (ix) the amino acid P at position 450, the amino acid P at position 451, and the amino acid L at position 452;
- (x) the amino acid P at position 450, the amino acid A at position 451, and the amino acid R at position 452;
- (xi) the amino acid S at position 450, the amino acid R at position 451, and the amino acid R at position 452;
- (xii) the amino acid F at position 450, the amino acid F at position 451, and the amino acid H at position 452;
- (xiii) the amino acid R at position 450, the amino acid F at position 451, and the amino acid S at position 452;
- (xiv) the amino acid Y at position 450, the amino acid S at position 451, and the amino acid M at position 452; or
- (xv) the amino acid P at position 450, the amino acid F at position 451, and the amino acid L at position 452;
- wherein (i)-(xv) is numbered according to SEQ ID NO: 138.
  
  316. The AAV capsid variant of any one of embodiments 300-315, which comprises:
- (i) the amino acid sequence of any one of SEQ ID NOs: 3849-3982, 2984-4010, 4681-4693;
- (ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 amino acids, e.g., consecutive amino acids, thereof;
- (iii) an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (i); or
- (iv) an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the amino acid sequences in (i).
  
  317. The AAV capsid variant of any one of embodiments 300-316, which does not comprise the amino acid sequence of GSGSPHSKAQNQQ (SEQ ID NO: 1801) or GSGSPHSKAQNQQT (SEQ ID NO: 200).
  
  318. The AAV capsid variant of any one of embodiments 300-317, wherein [A]-[B] is present in loop IV.
  
  319. The AAV capsid variant of any one of embodiments 300-318, wherein [A] is present immediately subsequent to position 452, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138 or 981.
  
  320. The AAV capsid variant of any one of embodiments 300-319, wherein [A] replaces positions 453-455 (e.g., G453, S454, G455), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138 or 981.
  
  321. The AAV capsid variant of any one of embodiments 300-320, wherein [A] is present immediately subsequent to position 452, and wherein [A] replaces positions 453-455 (e.g., G453, S454, G455), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138 or 981.
  
  322. The AAV capsid variant of any one of embodiments 300-321, wherein [B] is present immediately subsequent to [A].
  
  323. The AAV capsid variant of any one of embodiments 300-322, wherein [B] replaces positions 456 and 457 (e.g., Q456, N457), numbered relative to SEQ ID NO: 138.
  
  324. The AAV capsid variant of any one of embodiments 300-323, wherein [A]-[B] replaces positions 453-457 (e.g., G453, S454, G455, Q456, N457), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  325. The AAV capsid variant of any one of embodiments 300-324, wherein [A]-[B] is present immediately subsequent to position 452, and wherein [A]-[B] replaces positions 453-457 (e.g., G453, S454, G455, Q456, N457), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  326. The AAV capsid variant of any one of embodiments 300-325, which comprises from N-terminus to C-terminus, [A][B].
  
  327. The AAV capsid variant of any one of the preceding embodiments, which comprises at least one, two, three or four (e.g., from 1-4 to 1-5) charged amino acid residues (e.g., acidic and/or basic amino acid residues) relative to SEQ ID NO: 138, which is present N-terminal to the amino acid sequence of SPH (e.g., within 1, 2, 3, 4, 5, or 6 amino acids from the start of the SPH amino acid sequence (e.g., within positions 450-455 numbered according to SEQ ID NO: 138)), optionally wherein the amino acid sequence of SPH is present at positions 456-458 numbered according to any one of SEQ ID NOs: 36-59, 981, or 982.
  
  328. The AAV capsid variant of embodiment 327, wherein the amino acid sequence of SPH is present at positions 456-458 numbered according to any one of SEQ ID NOs: 36-59, 981, or 982.
  
  329. The AAV capsid variant of embodiment 327 or 328, which comprises less than four, less than three, less than two (e.g., two or one) charged amino acid residues (e.g., acidic and/or basic amino acid residues) relative to SEQ ID NO: 138.
  
  330. The AAV capsid variant of any one of embodiments 327-329, which comprises one charged amino acid residues (e.g., an acidic or basic amino acid residue) relative to SEQ ID NO: 138, optionally at any one of positions 450-455 numbered relative to SEQ ID NO: 138.
  
  331. The AAV capsid variant of any one of embodiments 327-330, wherein the charged amino acid residue is an acidic amino acid (e.g., D or E).
  
  332. The AAV capsid variant of any one of embodiments 327-331, wherein the charged amino acid residue is a negatively charged amino acid (e.g., D or E).
  
  333. The AAV capsid variant of any one of embodiments 327-332, wherein the charged amino acid residue is D.
  
  334. The AAV capsid variant of any one of embodiments 327-333, wherein the charged amino acid residue is E.
  
  335. The AAV capsid variant of any one of embodiments 327-334, wherein the charged amino acid residue is a basic amino acid (e.g., K, R, or H).
  
  336. The AAV capsid variant of any one of embodiments 327-335, wherein the charged amino acid residue is a positively charged amino acid (e.g., K, R, or H).
  
  337. The AAV capsid variant of any one of embodiments 327-336, wherein the charged amino acid residue is H.
  
  338. The AAV capsid variant of any one of embodiments 327-337, wherein the charged amino acid residue is R.
  
  339. The AAV capsid variant of any one of embodiments 327-338, wherein the charged amino acid residue is K.
  
  340. The AAV capsid variant of any one of embodiments 327-339, wherein the AAV capsid variant comprises an acidic amino acid (e.g., E or D) and a basic amino acid (e.g., R, K, or H).
  
  341. The AAV capsid variant of any one of embodiments 327-340, at least one, two, three or four charged amino acid residues is present within 1, 2, 3, 4, 5, or 6 (e.g., 1-6) amino acids from the start of the SPH amino acid sequence.
  
  342. The AAV capsid variant of any one of embodiments 327-341, which comprises two charged amino acid residue immediately preceding the amino acid sequence of SPH (e.g., at positions 454 and 455, numbered according to SEQ ID NO: 138 or SEQ ID NO: 982).
  
  343. The AAV capsid variant of any one of embodiments 327-342, which comprises a charged amino acid residue (e.g., E) within 1, 2, 3, 4, 5 (e.g., 5) amino acids from the start of the SPH amino acid sequence.
  
  344. The AAV capsid variant of any one of embodiments 327-343, which comprises a charged amino acid residue (e.g., E) at position 451, numbered according to any one of SEQ ID NO: 138, 981, or 982.
  
  345. The AAV capsid variant of any one of embodiments 327-344, which comprises E at position 451, numbered according to any one of SEQ ID NOs: 138, 981, or 982.
  
  346. The AAV capsid variant of any one of embodiments 327-345, which comprises a charged amino acid residue (e.g., R or K) at position 452, numbered according to any one of SEQ ID NOs: 138, 981, or 982.
  
  347. The AAV capsid variant of any one of embodiments 327-346, which comprises R at position 452, numbered according to SEQ ID NO: 138 or SEQ ID NO: 982.
  
  348. The AAV capsid variant of any one of embodiments 327-347, which comprises E at position 451 and R at position 452, numbered according to SEQ ID NO: 138 or SEQ ID NO: 982.
  
  349. The AAV capsid variant of any one of embodiments 327-348, which has decreased tropism for a liver cell or tissue, relative to the tropism of a reference sequence comprising the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981.
  
  350. The AAV capsid variant of any one of the preceding embodiments, which comprises at least one, two, three or four (e.g., from 1-4 to 1-5) charged amino acid residues (e.g., basic amino acid residues) relative to SEQ ID NO: 138, which is present C-terminal to the amino acid sequence of SPH (e.g., within 1, 2, 3, 4, 5, 6, or 7 amino acids from the end of the SPH amino acid sequence (e.g., within positions 459-465 numbered according to any one of SEQ ID NOs: 36-59, or 981)), optionally wherein the amino acid sequence of SPH is present at positions 456-458 numbered according to any one of SEQ ID NOs: 36-59, 981, or 982.
  
  351. The AAV capsid variant of embodiment 350, wherein the amino acid sequence of SPH is present at positions 456-458 numbered according to any one of SEQ ID NOs: 36-59, 981, or 982.
  
  352. The AAV capsid variant of embodiment 350 or 351, which comprises less than four, less than three, less than two (e.g., two or one) charged amino acid residues (e.g., basic amino acid residues) relative to SEQ ID NO: 138.
  
  353. The AAV capsid variant of any one of embodiments 350-352, which comprises one charged amino acid residues (e.g., an basic amino acid residue) relative to SEQ ID NO: 138, optionally at any one of positions 456-460 numbered relative to SEQ ID NO: 138 or at positions 462-466 numbered according to any one of SEQ ID NOs: 36-59, 981, or 982.
  
  354. The AAV capsid variant of any one of embodiments 350-353, wherein the charged amino acid residue is a basic amino acid (e.g., R or K).
  
  355. The AAV capsid variant of any one of embodiments 350-354, wherein the charged amino acid residue is a positively charged amino acid (e.g., R or K).
  
  356. The AAV capsid variant of any one of embodiments 350-355, wherein the charged amino acid residue is R.
  
  357. The AAV capsid variant of any one of embodiments 350-355, wherein the charged amino acid residue is K.
  
  358. The AAV capsid variant of any one of embodiments 350-357, at least one, two, three or four charged amino acid residues is present within 1, 2, 3, 4, 5, 6, 7 (e.g., 1-7) amino acids from the end of the SPH amino acid sequence.
  
  359. The AAV capsid variant of any one of embodiments 350-358, which comprises a charged amino acid residue (e.g., K or R) immediately after the SPH sequence (e.g., at position 459 numbered according to SEQ ID NO: 981).
  
  360. The AAV capsid variant of any one of embodiments 350-359, which comprises a charged amino acid residue (e.g., K or R) at position 459, numbered according to SEQ ID NO: 138 or SEQ ID NO: 982.
  
  361. The AAV capsid variant of any one of embodiments 350-360, which comprises K at position 459, numbered according to SEQ ID NO: 981.
  
  362. The AAV capsid variant of any one of embodiments 350-360, which comprises R at position 459, numbered according to SEQ ID NO: 981.
  
  363. The AAV capsid variant of any one of embodiments 350-362, which comprises a charged amino acid residue (e.g., R or K) at one, two three, four, five, or all of positions 460, 461, 462, 463, 464, and/or 465, numbered according to SEQ ID NO: 138 or 981.
  
  364. The AAV capsid variant of any one of embodiments 300-326 or 350-363, which has increased tropism for a liver cell or tissue, relative to the tropism of a reference sequence comprising the amino acid sequence of SEQ ID NO: 138.
  
  365. The AAV capsid variant of any one of embodiments 300-326 or 350-364, which is enriched at least about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 150, 160, 170, 180, 190, or 200-fold, in the liver compared to a reference sequence of SEQ ID NO: 138, e.g., when measured by an assay as described in Example 4.
  
  366. The AAV capsid variant of any one of embodiments 300-326, 364, or 365, which has reduced tropism for a CNS cell or tissue, e.g., a brain cell, brain tissue, spinal cord cell, or spinal cord tissue, relative to the tropism of a reference sequence comprising the amino acid sequence of SEQ ID NO: 138.
  
  367. The AAV capsid variant of any one of embodiments 300-326 or 364-366, which shows preferential transduction in a liver region relative to the transduction in the brain and/or dorsal root ganglia (DRG).
  
  368. The AAV capsid variant of any one of embodiments 300-326 or 364-367, which shows preferential transduction in a liver region relative to the transduction in the heart and/or muscle (e.g., quadriceps).
  
  369. An AAV capsid variant comprising:
- (a) the amino acid sequence of any of the sequences provided in Tables 1, 2A, 2B, 13-19;
- (b) an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 consecutive amino acids from any one of the sequences provided in Tables 1, 2A, 2B, 13-19; or
- (c) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids, relative to any one of the sequences provided in Tables 1, 2A, 2B, 13-19; or
- (d) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of any one of the sequences provided in Tables 1, 2A, 2B, 13-19.
  
  370. An AAV capsid variant comprising:
- (a) the amino acid sequence of any of SEQ ID NOs: 945-980 or 985-986;
- (b) an amino acid sequence comprising at least 3, 4, or 5 consecutive amino acids from any one of SEQ ID NOs: 945-980 or 985-986; or
- (c) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids, relative to the amino acid sequence of any one of SEQ ID NOs: 945-980 or 985-986;
- (d) an amino sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of any one of SEQ ID NOs: 945-980 or 985-986.
  
  371. An AAV capsid variant comprising:
- (a) the amino acid sequence of any of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909;
- (b) an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 consecutive amino acids from any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909;
- (c) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids, relative to the amino acid sequence of any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909; or
- (d) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909.
  
  372. An AAV capsid variant comprising:
- (a) the amino acid sequence of any of SEQ ID NOs: 3849-4051 or 4681-4693;
- (b) an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17 consecutive amino acids from any one of SEQ ID NOs: 3849-4051 or 4681-4693;
- (c) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids, relative to the amino acid sequence of any one of SEQ ID NOs: 3849-4051 or 4681-4693; or
- (d) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of any one of SEQ ID NOs: 3849-4051 or 4681-4693.
  
  373. An AAV capsid variant comprising:
- (a) the amino acid sequence of any of SEQ ID NOs: 4052-4092;
- (b) an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17 consecutive amino acids from any one of SEQ ID NOs: 4052-4092;
- (c) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids, relative to the amino acid sequence of any one of SEQ ID NOs: 4052-4092; or
- (d) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of any one of SEQ ID NOs: 4052-4092.
  
  374. An AAV capsid variant comprising:
- (a) the amino acid sequence of any of SEQ ID NOs: 4056, 4058, 4059, 4062-4064, 4066, 4067, 4080, 4084, 4090, or 4095-4097;
- (b) an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17 consecutive amino acids from any one of SEQ ID NOs: 4056, 4058, 4059, 4062-4064, 4066, 4067, 4080, 4084, 4090, or 4095-4097;
- (c) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids, relative to the amino acid sequence of any one of SEQ ID NOs: 4056, 4058, 4059, 4062-4064, 4066, 4067, 4080, 4084, 4090, or 4095-4097; or
- (d) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of any one of SEQ ID NOs: 4056, 4058, 4059, 4062-4064, 4066, 4067, 4080, 4084, 4090, or 4095-4097.
  
  375. The AAV capsid variant of embodiment 369 or 371, comprising an amino acid sequence comprising at least 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 consecutive amino acids from any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909.
  
  376. The AAV capsid variant of any one of embodiments 369-375, wherein the 3 consecutive amino acids comprise SPH.
  
  377. The AAV capsid variant of any one of embodiments 369-371 or 376, wherein the 4 consecutive amino acids comprise SPHS (SEQ ID NO: 4700).
  
  378. The AAV capsid variant of any one of embodiments 369-371, 376, or 377, wherein the 5 consecutive amino acids comprise SPHSK (SEQ ID NO: 4701).
  
  379. The AAV capsid variant of any one of embodiments 369-371 or 376-378, wherein the 6 consecutive amino acids comprise SPHSKA (SEQ ID NO: 941).
  
  380. The AAV capsid variant of embodiment 369-371, wherein the 3 consecutive amino acids comprise HDS.
  
  381. The AAV capsid variant of any one of embodiments 369-371 or 380, wherein the 4 consecutive amino acids comprise HDSP (SEQ ID NO: 4702).
  
  382. The AAV capsid variant of any one of embodiments 369-371, 380, or 381, wherein the 5 consecutive amino acids comprise HDSPH (SEQ ID NO: 4703).
  
  383. The AAV capsid variant of any one of embodiments 369-371 or 380-382, wherein the 6 consecutive amino acids comprise HDSPHK (SEQ ID NO: 2).
  
  384. The AAV capsid variant of any one of embodiments 369-371, wherein:
- (i) the 3 consecutive amino acids comprise SPH; (ii) the 4 consecutive amino acids comprise SPHK (SEQ ID NO: 6398);
- (iii) the 5 consecutive amino acids comprise SPHKY (SEQ ID NO: 4715); and/or
- (iv) the 6 consecutive amino acids comprise SPHKYG (SEQ ID NO: 966).
  
  385. The AAV capsid variant of embodiment 369 or 371, which comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909.
  
  386. The AAV capsid variant of any one of embodiments 369,371, or 385, which comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941).
  
  387. The AAV capsid variant of any one of embodiments 369, 371, or 385, which comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2).
  
  388. The AAV capsid variant of any one of embodiments 369-371, 384, or 385, which comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of SPHKYG (SEQ ID NO: 966).
  
  389. The AAV capsid variant of embodiment 370, which comprises:
- (i) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589);
- (ii) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of KAEIGHDSPHKSGQNQQT (SEQ ID NO: 1754)
- (iii) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of KTEKMSGSPHSKAQNQQT (SEQ ID NO: 3241);
- (iv) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of KTINGHDSPHSKAQNLQT (SEQ ID NO: 4100); or
- (v) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of KTVNGHDSPHSKAQNQQT (SEQ ID NO: 4062).
  
  390. The AAV capsid variant of embodiment 369 or 371, which comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909.
  
  391. The AAV capsid variant of any one of embodiments 369,371, or 390, which comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941).
  
  392. The AAV capsid variant of any one of embodiments 369, 371, or 390, which comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2).
  
  393. The AAV capsid variant of any one of embodiments 369, 371, 384, or 390, which comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of SPHKYG (SEQ ID NO: 966).
  
  394. The AAV capsid variant of embodiment 369, which comprises:
- (i) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589);
- (ii) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of KAEIGHDSPHKSGQNQQT (SEQ ID NO: 1754);
- (iii) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of KTEKMSGSPHSKAQNQQT (SEQ ID NO: 3241);
- (iv) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of KTINGHDSPHSKAQNLQT (SEQ ID NO: 4100); or
- (v) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of KTVNGHDSPHSKAQNQQT (SEQ ID NO: 4062).
  
  395. The AAV capsid variant of any one of embodiments 1-129, 269, 271, 375-388, or 390-394, which comprises the amino acid sequence of any of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909.
  
  396. The AAV capsid variant of any one of embodiments 295-297, 301-305, 313, 314, 318, 319, or 323, which comprises the amino acid sequence of ERVSGSPHSKA (SEQ ID NO: 6399), optionally wherein the amino acid sequence is present immediately subsequent to position 450 and replaces positions 451-455 (e.g., 1451, N542, G453, S454, G455), numbered according to SEQ ID NO: 138.
  
  397. The AAV capsid variant of any one of embodiments 369-371, 375-379, 385, 386, 389-391, or 394-396, which comprises the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589), optionally wherein the amino acid sequence is present immediately subsequent to position 448 and replaces positions 449-460 (e.g., K449, T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, Q459, T460), numbered according to SEQ ID NO: 138.
  
  398. The AAV capsid variant of any one of embodiments 269-371, 375, 380-383, 385, 387, 389, 390, 393, or 394, which comprises the amino acid sequence of AEIGHDSPHKSG (SEQ ID NO: 6400), optionally wherein the amino acid sequence is present immediately subsequent to position 449 and replaces positions 450-455 (e.g., T450, 1451, N452, G453, S454, G455), numbered according to SEQ ID NO: 138.
  
  399. The AAV capsid variant of any one of embodiments 369-371, 375, 380-383, 385, 387, 389, 390, 393, 394, or 398, which comprises the amino acid sequence of KAEIGHDSPHKSGQNQQT (SEQ ID NO: 1754), optionally wherein the amino acid sequence is present immediately subsequent to position 448 and replaces positions 449-460 (e.g., K449, T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, Q459, T460), numbered according to SEQ ID NO: 138.
  
  400. The AAV capsid variant of any one of embodiments 369-371, 375-379, 390, 391, or 395, which comprises the amino acid sequence of EKMSGSPHSKA (SEQ ID NO: 6401), optionally wherein the amino acid sequence is present immediately subsequent to position 450 and replaces positions 451-455 (e.g., 1451, N452, G453, S454, G455), numbered according to SEQ ID NO: 138.
  
  401. The AAV capsid variant of any one of embodiments 369-371, 375-379, 390, 391, or 395, which comprises the amino acid sequence of KTEKMSGSPHSKAQNQQT (SEQ ID NO: 3241), optionally wherein the amino acid sequence is present immediately subsequent to position 448 and replaces positions 449-460 (e.g., K449, T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, Q459, T460), numbered according to SEQ ID NO: 138.
  
  402. The AAV capsid variant of any one of embodiments 369-371, 375-379, 390, 391, or 395, which comprises the amino acid sequence of HDSPHSKAQNL (SEQ ID NO: 6402), optionally wherein the amino acid sequence is present immediately subsequent to position 453 and replaces positions 456-458 (e.g., Q456, N457, Q458), numbered according to SEQ ID NO: 138.
  
  403. The AAV capsid variant of any one of embodiments 369-371, 375-379, 390, 391, or 395, which comprises the amino acid sequence of KTINGHDSPHSKAQNLQT (SEQ ID NO: 4100), optionally wherein the amino acid sequence is present immediately subsequent to position 448 and replaces positions 449-460 (e.g., K449, T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, Q459, T460), numbered according to SEQ ID NO: 138.
  
  404. The AAV capsid variant of any one of embodiments 369-371, 375-379, 390, 391, or 395, which comprises the amino acid sequence of VNGHDSPHSKA (SEQ ID NO: 6403), optionally wherein the amino acid sequence is present immediately subsequent to position 450 and replaces positions 451-455 (e.g., 1451, N452, G453, S454, G455), numbered according to SEQ ID NO: 138.
  
  405. The AAV capsid variant of any one of embodiments 369-371, 375-379, 390, 391, or 395, which comprises the amino acid sequence of KTVNGHDSPHSKAQNQQT (SEQ ID NO: 4062), optionally wherein the amino acid sequence is present immediately subsequent to position 448 and replaces positions 449-460 (e.g., K449, T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, Q459, T460), numbered according to SEQ ID NO: 138.
  
  406. The AAV capsid variant of any one of embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, or 395, comprising an amino acid sequence encoded by: the nucleotide sequence of SEQ ID NO: 942; a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of SEQ ID NO: 942; or a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides relative to the nucleotide sequence of SEQ ID NO: 942.
  
  407. The AAV capsid variant of any one of embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, or 395, comprising an amino acid sequence encoded by: the nucleotide sequence of SEQ ID NO: 3; a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of SEQ ID NO: 3; or a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides relative to the nucleotide sequence of SEQ ID NO: 3.
  
  408. The AAV capsid variant of any one of embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, or 406, wherein the nucleotide sequence encoding the capsid variant comprises the nucleotide sequence of SEQ ID NO: 942; a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of SEQ ID NO: 942; or a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides relative to the nucleotide sequence of SEQ ID NO: 942.
  
  409. The AAV capsid variant of any one of embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, or 407, wherein the nucleotide sequence encoding the capsid variant comprises the nucleotide sequence of SEQ ID NO: 3; a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of SEQ ID NO: 3; or a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides relative to the nucleotide sequence of SEQ ID NO: 3.
  
  410. The AAV capsid variant of any one of embodiments 369-409, wherein the amino acid sequence is present in loop IV.
  
  411. The AAV capsid variant of any one of embodiments 369-410, wherein the amino acid sequence is present immediately subsequent to position 448, 449, 450, 451, 452, 453, 454, or 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  412. The AAV capsid variant of any one of embodiments 369-411, wherein the amino acid sequence replaces amino acids 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, and/or 460 (e.g., K449, T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, Q459, and/or T460), numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  413. The AAV capsid variant of any one of embodiments 369-412, wherein the amino acid sequence is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  414. The AAV capsid variant of any one of embodiments 369-413, wherein the amino acid sequence is present immediately subsequent to position 453, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  415. The AAV capsid variant of any one of embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, or 410-413, comprising the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  416. The AAV capsid variant of any one of embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, or 415, comprising the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981.
  
  417. The AAV capsid variant of embodiment 415 or 416, which further comprises an amino acid other than I at position 451, an amino acid other than N at position 452, and an amino acid other than G at position 453, numbered according to any one of SEQ ID NOs: 36, 138, or 981.
  
  418. The AAV capsid variant of any one of embodiments 415-417, which further comprises E at position 451, R at position 452, and V at position 453, numbered according to any one of SEQ ID NOs: 36, 138, or 981.
  
  419. The AAV capsid variant of any one of embodiments 415-418, which further comprises the substitutions 1451E, N452R, and G453V, numbered according to any one of SEQ ID NOs: 36, 138, or 981.
  
  420. The AAV capsid variant of any one of embodiments 415-419, which comprises:
- (i) E at position 451, R at position 452, and V at position 453, numbered according to any one of SEQ ID NOs: 36, 138, or 981; and
- (ii) the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to position 455, numbered according to any one of SEQ ID NOs: 36, 138, or 981.
  
  421. The AAV capsid variant of embodiment 415 or 416, which further comprises an amino acid other than I at position 451, an amino acid other than N at position 452, and/or G at position 453, numbered according to SEQ ID NO: 39 or 138.
  
  422. The AAV capsid variant of any one of embodiments 415, 416, or 421, which further comprises E at position 451, K at position 452, and/or M at position 453, numbered according to SEQ ID NO: 138 or 39.
  
  423. The AAV capsid variant of any one of embodiments 415, 416, 421, or 422, which further comprises the substitutions I451E, N452K, and G453M, numbered according to SEQ ID NO: 39 or 138.
  
  424. The AAV capsid variant of any one of embodiments 415, 416, or 421-423, which comprises:
- (i) E at position 451, K at position 452, and M at position 453, numbered according to SEQ ID NO: 39 or 138; and
- (ii) the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to position 455, numbered according to SEQ ID NO: 39 or 138.
  
  425. The AAV capsid variant of embodiment 415 or 416, which further comprises an amino acid other than S at position 454, an amino acid other than G at position 455, and/or Q at position 458, numbered according to SEQ ID NO: 138.
  
  426. The AAV capsid variant of any one of embodiments 415, 416, or 425, which further comprises H at position 454, D at position 455, and/or L at position 458, numbered according to SEQ ID NO: 138.
  
  427. The AAV capsid variant of any one of embodiments 415, 416, 425, or 426, which further comprises the substitutions S454H, G455D, and Q458L, numbered according to SEQ ID NO: 138.
  
  428. The AAV capsid variant of any one of embodiments 415, 416, or 425-427, which comprises:
- (i) H at position 454, D at position 455, and/or L at position 458, numbered according to SEQ ID NO: 138; and
- (ii) the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to position 455, numbered according to SEQ ID NO: 138.
  
  429. The AAV capsid variant of embodiment 415 or 416, which further comprises an amino acid other than I at position 451, an amino acid other than S at position 454, and/or an amino acid other than G at position 455, numbered according to SEQ ID NO: 52 or 138.
  
  430. The AAV capsid variant of any one of embodiments 415, 416, or 429, which further comprises V at position 451, H at position 454, and/or D at position 455, numbered according to SEQ ID NO: 52 or 138.
  
  431. The AAV capsid variant of any one of embodiments 415, 416, 429, or 430, which further comprises the substitutions 1451V, S454H, and/or G455D, numbered according to SEQ ID NO: 52 or 138.
  
  432. The AAV capsid variant of any one of embodiments 415, 416, or 429-431, which comprises:
- (i) V at position 451, H at position 454, and/or D at position 455, numbered according to SEQ ID NO: 52 or 138; and
- (ii) the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to position 455, numbered according to SEQ ID NO: 52 or 138.
  
  433. The AAV capsid variant of any one of embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, or 414, comprising the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein the amino acid sequence is present immediately subsequent to position 453, numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  434. The AAV capsid variant of any one of embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, or 433, comprising the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein the amino acid sequence is present immediately subsequent to position 453, numbered according to the amino acid sequence of SEQ ID NO: 982.
  
  435. The AAV capsid variant of any one of embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433, or 434, comprising the amino acid sequence of SPHKSG (SEQ ID NO: 946), wherein the amino acid sequence is present immediately subsequent to position 455, numbered relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 982.
  
  436. The AAV capsid variant of any one of embodiments 369-435, which comprises:
- (i) the amino acid sequence of HDSPHSKA (SEQ ID NO: 4486), which is present immediately subsequent to position 453; and
- (ii) a deletion of amino acids SG at position 454 and 455;
- wherein (i) and (ii) are numbered according to SEQ ID NO: 138.
  
  437. The AAV capsid variant of any one of embodiments 369-436, which comprises the amino acids HD at position 454 and 455, and further comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941), which is present immediately subsequent to position 455, numbered relative to SEQ ID NO: 138.
  
  438. The AAV capsid variant of any one of embodiments 433-435, which further comprises an amino acid other than T at position 450, an amino acid other than I at position 451, and an amino acid other than N at position 452, numbered according to SEQ ID NO: 138 or 982.
  
  439. The AAV capsid variant of any one of embodiments 433-435 or 438, which further comprises A at position 450, E at position 451, and I at position 452, numbered according to SEQ ID NO: 138 or 982.
  
  440. The AAV capsid variant of any one of embodiments 433-435, 438, or 439, which further comprises the substitutions T450A, I451E, and N452I, numbered according to SEQ ID NO: 138 or 982.
  
  441. The AAV capsid variant of any one of embodiments 433, 434, or 438-440, which comprises:
- (i) A at position 450, E at position 451, and I at position 452, numbered according to SEQ ID NO: 138 or 982; and
- (ii) the amino acid sequence of HDSPHK (SEQ ID NO: 2), which is present immediately subsequent to positions 453, numbered according to SEQ ID NO: 138 or 982.
  
  442. The AAV capsid variant of any one of embodiments 1-22, 25-27, 31, 34-42, 45-50, 53-63, 69, 79, 83-86, 91-98, 102, 103, 110, 111, 118-129, 369-371, 384, 385, 390, 393, 395, 410-413, comprising the amino acid sequence of SPHKYG (SEQ ID NO: 966), wherein the amino acid sequence is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  443. An AAV capsid variant comprising the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein the amino acid sequence is present immediately subsequent to position 453, numbered according to the amino acid sequence of SEQ ID NO: 982.
  
  444. An AAV capsid variant comprising the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to position 455, numbered according to the amino acid sequence of SEQ ID NO: 981.
  
  445. An AAV capsid variant comprising the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein the amino acid sequence is present immediately subsequent to position 453, numbered according to the amino acid sequence of SEQ ID NO: 37, and optionally further comprising:
- (i) one, two, or all of an amino acid other than T at position 450, an amino acid other than I at position 541, and/or an amino acid other than N at position 452, numbered according to SEQ ID NO: 138 or 37;
- (ii) one, two, or all of A at position 450, E at position 451, and/or I at position 452, numbered according to SEQ ID NO: 138 or 37.
  
  446. An AAV capsid variant comprising the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to position 455, numbered according to the amino acid sequence of any one of SEQ ID NO: 36, 38-55, 57, or 59.
  
  447. The AAV capsid variant of any one of the preceding embodiments, which further comprises:
- (i) a modification, e.g., an insertion, substitution (e.g., conservative substitution), and/or deletion, in loop I, II, VI and/or VIII; and/or
- (ii) a substitution at position K449, e.g., a K449R substitution, numbered according to SEQ ID NO: 138.
  
  448. The AAV capsid variant of any one of the preceding embodiments, which comprises an amino acid sequence comprising at least one, two or three modifications, e.g., substitutions (e.g., conservative substitutions), but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of SEQ ID NO: 138.
  
  449. The AAV capsid variant of any one of the preceding embodiments, which comprises an amino acid sequence comprising at least one, two or three, but no more than 30, 20 or 10 different amino acids relative to the amino acid sequence of SEQ ID NO: 138.
  
  450. The AAV capsid variant of any one of the preceding embodiments, which comprises the amino acid sequence of SEQ ID NO: 138, or an amino acid sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  451. The AAV capsid variant of any one of the preceding embodiments, which comprises the amino acid sequence of SEQ ID NO: 138.
  
  452. The AAV capsid variant of any one of the preceding embodiments, which comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 137, or a sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  453. The AAV capsid variant of any one of the preceding embodiments, wherein the nucleotide sequence encoding the capsid variant comprises the nucleotide sequence of SEQ ID NO: 137, or a sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  454. The AAV capsid variant of any one of the preceding embodiments, which comprises a VP1 protein, a VP2 protein, a VP3 protein, or a combination thereof.
  
  455. The AAV capsid variant of any one of embodiments 1-454, which comprises the amino acid sequence corresponding to positions 138-742, e.g., a VP2, of SEQ ID NO: 981 or 982, or a sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  456. The AAV capsid variant of any one of embodiments 1-455, which comprises the amino acid sequence corresponding to positions 203-742, e.g., a VP3, of SEQ ID NO: 981 or 982, or a sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  457. The AAV capsid variant of any one of embodiments 1-456, which comprises the amino acid sequence corresponding to positions 138-736, e.g., a VP2, of SEQ ID NO: 138, or a sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  458. The AAV capsid variant of any one of embodiments 1-457, which comprises the amino acid sequence corresponding to positions 203-736, e.g., a VP3, of SEQ ID NO: 138, or a sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  459. The AAV capsid variant of any one of embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, or 446-458, comprising an amino acid sequence comprising at least 3, 4, 5, or 6 consecutive amino acids from the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein:
- (i) the 3 consecutive amino acids comprise SPH;
- (ii) the 4 consecutive amino acids comprise SPHS (SEQ ID NO: 4700);
- (iii) the 5 consecutive amino acids comprise SPHSK (SEQ ID NO: 4701); or
- (iv) the 6 consecutive amino acids comprise SPHSKA (SEQ ID NO: 941);
- wherein the AAV capsid variant comprises: (a) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981; (b) a VP2 protein comprising the amino acid sequence of positions 138-736 of SEQ ID NO: 138 or positions 138-742 of SEQ ID NO: 981; (c) a VP3 protein comprising the amino acid sequence of positions 203-736 of SEQ ID NO: 138 or positions 203-742 of SEQ ID NO: 981; or (d) an amino acid sequence with at least 90% (e.g., at least about 95, 96, 97, 98, or 99%) sequence identity to any of the amino acid sequences in (a)-(c).
  
  460. The AAV capsid variant of any one of embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, or 446-459, comprising an amino acid sequence comprising at least 3, 4, 5, or 6 consecutive amino acids from the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein:
- (i) the 3 consecutive amino acids comprise SPH;
- (ii) the 4 consecutive amino acids comprise SPHS (SEQ ID NO: 4700);
- (iii) the 5 consecutive amino acids comprise SPHSK (SEQ ID NO: 4701); or
- (iv) the 6 consecutive amino acids comprise SPHSKA (SEQ ID NO: 941);
- wherein the AAV capsid variant comprises an amino acid sequence at least 90% (e.g., at least about 95, 96, 97, 98, or 99%) identical to the amino acid sequence of SEQ ID NO: 981.
  
  461. The AAV capsid variant of any one of embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, or 446-460, comprising one or two, but no more than three substitutions relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the AAV capsid variant comprises:
- (a) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981;
- (b) a VP2 protein comprising the amino acid sequence of positions 138-736 of SEQ ID NO: 138 or positions 138-742 of SEQ ID NO: 981;
- (c) a VP3 protein comprising the amino acid sequence of positions 203-736 of SEQ ID NO: 138 or positions 203-742 of SEQ ID NO: 981; or
- (d) an amino acid sequence with at least 90% (e.g., at least about 95, 96, 97, 98, or 99%) sequence identity to any of the amino acid sequences in (a)-(c).
  
  462. The AAV capsid variant of any one of embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, or 446-461, comprising one or two, but no more than three substitutions relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the AAV capsid variant comprises an amino acid sequence at least 90% (e.g., at least about 95, 96, 97, 98, or 99%) identical to the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981.
  
  463. The AAV capsid variant of any one of embodiments 459-462, wherein the amino acid sequence is present immediately subsequent to position 455, numbered according to SEQ ID NO: 138 or 981.
  
  464. The AAV capsid variant of any one of embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, or 447-458, comprising an amino acid sequence comprising at least 3, 4, 5, or 6 consecutive amino acids from the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein:
- (i) the 3 consecutive amino acids comprise HDS;
- (ii) the 4 consecutive amino acids comprise HDSP (SEQ ID NO: 4702);
- (iii) the 5 consecutive amino acids comprise HDSPH (SEQ ID NO: 4703); or
- (iv) the 6 consecutive amino acids comprise HDSPHK (SEQ ID NO: 2);
- wherein the AAV capsid variant comprises: (a) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 982; (b) a VP2 protein comprising the amino acid sequence of positions 138-736 of SEQ ID NO: 138 or positions 138-742 of SEQ ID NO: 982; (c) a VP3 protein comprising the amino acid sequence of positions 203-736 of SEQ ID NO: 138 or positions 203-742 of SEQ ID NO: 982; or (d) an amino acid sequence with at least 90% (e.g., at least about 95, 96, 97, 98, or 99%) sequence identity to any of the amino acid sequences in (a)-(c).
  
  465. The AAV capsid variant of any one of embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, or 464, comprising an amino acid sequence comprising at least 3, 4, 5, or 6 consecutive amino acids from the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein:
- (i) the 3 consecutive amino acids comprise HDS;
- (ii) the 4 consecutive amino acids comprise HDSP (SEQ ID NO: 4702);
- (iii) the 5 consecutive amino acids comprise HDSPH (SEQ ID NO: 4703); or
- (iv) the 6 consecutive amino acids comprise HDSPHK (SEQ ID NO: 2);
- wherein the AAV capsid variant comprises an amino acid sequence at least 90% (e.g., at least about 95, 96, 97, 98, or 99%) identical to the amino acid sequence of SEQ ID NO: 982.
  
  466. The AAV capsid variant of any one of embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, 464, or 465, comprising one or two, but no more than three substitutions relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein the AAV capsid variant comprises:
- (a) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 982;
- (b) a VP2 protein comprising the amino acid sequence of positions 138-736 of SEQ ID NO: 138 or positions 138-742 of SEQ ID NO: 982;
- (c) a VP3 protein comprising the amino acid sequence of positions 203-736 of SEQ ID NO: 138 or positions 203-742 of SEQ ID NO: 982; or
- (d) an amino acid sequence with at least 90% (e.g., at least about 95, 96, 97, 98, or 99%) sequence identity to any of the amino acid sequences in (a)-(c).
  
  467. The AAV capsid variant of any one of embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, or 464-466, comprising one or two, but no more than three substitutions relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein the AAV capsid variant comprises an amino acid sequence at least 90% (e.g., at least about 95, 96, 97, 98, or 99%) identical to the amino acid sequence of SEQ ID NO: 982.
  
  468. The AAV capsid variant of any one of embodiments 464-468, wherein the amino acid sequence is present immediately subsequent to position 453, numbered according to SEQ ID NO: 138 or 982.
  
  469. The AAV capsid variant of any one of embodiments 1-468, which comprises the amino acid sequence of any one of SEQ ID NO: 981 or 982, or an amino acid sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  470. The AAV capsid variant of any one of embodiments 1-469, which comprises an amino acid sequence comprising at least one, two or three modifications, e.g., substitutions (e.g., conservative substitutions), but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of SEQ ID NO: 981 or 982.
  
  471. The AAV capsid variant of any one of embodiments, 1-470, which comprises an amino acid sequence comprising at least one, two or three, but not more than 30, 20 or 10 different amino acids, relative to the amino acid sequence of SEQ ID NO: 981 or 982.
  
  472. The AAV capsid variant of any one of embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, 446-463, or 469-471, which comprises the amino acid sequence of SEQ ID NO: 981, or an amino acid sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  473. The AAV capsid variant of any one of embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, 446-463, or 469-472, which comprises an amino acid sequence comprising at least one, two or three modifications, e.g., substitutions (e.g., conservative substitutions), but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of SEQ ID NO: 981.
  
  474. The AAV capsid variant of any one of embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, 446-463, or 469-473, which comprises an amino acid sequence comprising at least one, two or three, but not more than 30, 20 or 10 different amino acids, relative to the amino acid sequence of SEQ ID NO: 981.
  
  475. The AAV capsid variant of any one of embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, or 464-471, which comprises the amino acid sequence of SEQ ID NO: 982, or an amino acid sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  476. The AAV capsid variant of any one of embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, 464-471, or 475, which comprises an amino acid sequence comprising at least one, two or three modifications, e.g., substitutions (e.g., conservative substitutions), but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of SEQ ID NO: 982.
  
  477. The AAV capsid variant of any one of embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, 464-471, 475, or 476, which comprises an amino acid sequence comprising at least one, two or three, but not more than 30, 20 or 10 different amino acids, relative to the amino acid sequence of SEQ ID NO: 982.
  
  478. The AAV capsid variant of any one of embodiments 1-477, which comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 983 or 984, or a nucleotide sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  479. The AAV capsid variant of any one of the preceding embodiments 1-478, wherein the nucleotide sequence encoding the capsid variant comprises the nucleotide sequence of SEQ ID NOs: 983 or 984, or a nucleotide sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  480. The AAV capsid variant of any one of the preceding embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, 446-463, 469-474, 478, or 479, wherein the nucleotide sequence encoding the capsid variant comprises the nucleotide sequence of SEQ ID NO: 983, or a nucleotide sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  481. The AAV capsid variant of any one of the preceding embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, 464-471, or 475-479, wherein the nucleotide sequence encoding the capsid variant comprises the nucleotide sequence of SEQ ID NO: 984, or a nucleotide sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  482. The AAV capsid variant of any one of the preceding embodiments, wherein the nucleotide sequence encoding the capsid variant is codon optimized.
  
  483. An AAV capsid variant, comprising the amino acid sequence of any one of embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, 446-463, 469-474, 478-480, or 482, and further comprising an amino acid sequence at least 95% identical to SEQ ID NO: 981.
  
  484. An AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 981.
  
  485. The AAV capsid variant of embodiment 483 or 484, wherein the nucleotide sequence encoding the AAV capsid variant comprises the nucleotide sequence of SEQ ID NO: 983, or a nucleotide sequence at least 90%, 95%, or 99% identical thereto.
  
  486. An AAV capsid variant, comprising the amino acid sequence of any one of embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, 464-471, 475-479, 481, or 482, and further comprising an amino acid sequence at least 95% identical to SEQ ID NO: 982.
  
  487. An AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 982.
  
  488. The AAV capsid variant of embodiment 486 or 487, wherein the nucleotide sequence encoding the AAV capsid variant comprises the nucleotide sequence of SEQ ID NO: 984, or a nucleotide sequence at least 90%, 95%, or 99% identical thereto.
  
  489. An AAV capsid variant comprising an amino acid sequence encoded by the nucleotide sequence of SEQ ID NOs: 983 or 984, or a nucleotide sequence at least 95% identical thereto.
  
  490. An AAV capsid variant comprising the amino acid sequence of any one of SEQ ID NOs: 36-59.
  
  491. An AAV capsid variant comprising an amino acid sequence encoded by the nucleotide sequence of any one of SEQ ID NOs: 12-35, or a nucleotide sequence at least 95% identical thereto.
  
  492. The AAV capsid variant of 490 or 491, wherein the nucleotide sequence encoding the AAV capsid variant comprises the nucleotide sequence of any one of SEQ ID NOs: 12-35, or a nucleotide sequence at least 95% identical thereto.
  
  493. The AAV capsid variant of any one of embodiments 1-299, 369-371, or 375-492, which has an increased tropism for a CNS cell or tissue, e.g., a brain cell, brain tissue, spinal cord cell, or spinal cord tissue, relative to the tropism of a reference sequence comprising the amino acid sequence of SEQ ID NO: 138.
  
  494. The AAV capsid variant of any one of embodiments 1-129, 168-299, 369-371, or 375-493, which transduces a brain region, e.g., a midbrain region (e.g., the hippocampus, or thalamus) or the brain stem, optionally wherein the level of transduction is at least 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, or 65-fold greater as compared to a reference sequence of SEQ ID NO: 138, e.g., when measured by an assay, e.g., an immunohistochemistry assay or a qPCR assay, e.g., as described in Example 2.
  
  495. The AAV capsid variant of any one of embodiments 1-129, 168-299, 369-371, or 375-494, which transduces a brain region, e.g., a midbrain region (e.g., the hippocampus, or thalamus) or the brain stem, optionally wherein the level of transduction is at least 30, 35, 40, 45, 50, 55, 60, or 65-fold greater as compared to a reference sequence of SEQ ID NO: 138, e.g., when measured by an assay, e.g., an immunohistochemistry assay or a qPCR assay, e.g., as described in Example 2.
  
  496. The AAV capsid variant of any one of embodiments 1-129, 168-299, 369-371, or 375-495, which is enriched at least about 3, 4, 5, 6, 7, 8, 9, or 10-fold, in the brain compared to a reference sequence of SEQ ID NO: 138, e.g., when measured by an assay as described in Example 1.
  
  497. The AAV capsid variant of any one of embodiments 1-129, 168-299, 369-371, or 375-496, which is enriched at least about 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80 or 85-fold, in the brain compared to a reference sequence of SEQ ID NO: 138, e.g., when measured by an assay as described in Example 1.
  
  498. The AAV capsid variant of any one of embodiments 1-129, 168-299, 369-371, or 375-497, which is enriched in the brain of at least two to three species, e.g., a non-human primate and rodent (e.g., mouse), e.g., as compared to a reference sequence of SEQ ID NO: 138.
  
  499. The AAV capsid variant of any one of embodiments 1-129, 168-299, 369-371, or 375-498, which is enriched at least about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 190, 200, 205, or 210-fold, in the brain of at least two to three species, e.g., a non-human primate and rodent (e.g., mouse), compared to a reference sequence of SEQ ID NO: 138, e.g., when measured by an assay as described in Example 1 or 5.
  
  500. The AAV capsid variant of embodiment 498 or 499, wherein the at least two to three species are Macaca fascicularis, Chlorocebus sabaeus, Callithrix jacchus, and/or mouse (e.g., BALB/c mice, C57B1/6 mice, and/or CD-1 outbred mice).
  
  501. The AAV capsid variant of any one of embodiments 130-146, 369, 410-414, 447-454, 457, 458, 482, or 493, which is enriched at least about 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, or 8-fold, in the brain compared to a reference sequence of SEQ ID NO: 981, e.g., when measured by an assay as described in Example 3.
  
  502. The AAV capsid variant of any one of embodiments 147-167, 369, 410-414, 447-454, 457, 458, 482, or 493, which is enriched at least about 2, 2.5, 3, 3.5, 4, 4.5, 5, or 5.5-fold, in the brain compared to a reference sequence of SEQ ID NO: 982, e.g., when measured by an assay as described in Example 3.
  
  503. The AAV capsid variant of any one of embodiments 1-129, 168-299, 369-371, or 375-500, which delivers an increased level of a payload to a brain region, optionally wherein the level of the payload is increased by at least 10, 12, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, or 70-fold, as compared to a reference sequence of SEQ ID NO: 138, e.g., when measured by an assay, e.g., a qRT-PCR or a qPCR assay (e.g., as described in Example 2 or 8).
  
  504. The AAV capsid variant of any one of embodiments 1-129, 168-299, 369-371, 375-500, or 503, which delivers an increased level of viral genomes to a brain region, optionally wherein the level of viral genomes is increased by at least 5, 10, 15, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50-fold, as compared to a reference sequence of SEQ ID NO: 138, e.g., when measured by an assay, e.g., a qRT-PCR or a qPCR assay (e.g., as described in Example 2 or 8).
  
  505. The AAV capsid variant of embodiment 504 or 504, wherein the brain region is a midbrain region (e.g., the hippocampus or thalamus), frontal cortex, temporal cortex, motor cortex, cerebral cortex, caudate, putamen, dentate nucleus, substantia nigra, or the brainstem.
  
  506. The AAV capsid variant of any one of embodiments 1-129, 168-299, 369-371, 375-500, or 503-505, which is enriched at least about 4, 5, 10, 15, 20, 25, 30, or 35-fold, in the spinal cord compared to a reference sequence of SEQ ID NO: 138, e.g., when measured by an assay as described in Example 1 or 8, optionally wherein the region of the spinal cord is a thoracic spinal cord region, cervical spinal cord region, C5 ventral horn region, lumbar spinal cord region, or L5 ventral horn region.
  
  507. The AAV capsid variant of any one of embodiments 1-129, 168-299, 369-371, 375-500, or 503-506, which shows preferential transduction in a brain region relative to the transduction in the dorsal root ganglia (DRG).
  
  508. The AAV capsid variant of any one of embodiments 1-129, 168-299, 369-371, 375-500, or 503-507, which shows preferential transduction in a brain region relative to the liver.
  
  509. The AAV capsid variant of any one of embodiments 1-129, 168-299, 369-371, 375-500, or 503-508, which shows preferential transduction in a brain region relative to the transduction in the heart.
  
  510. The AAV capsid variant of any one of embodiments 1-129, 168-299, 369-371, 375-500, or 503-509, which shows preferential transduction in a brain region relative to the transduction in the dorsal root ganglia (DRG) and the heart.
  
  511. The AAV capsid variant of any one of the preceding embodiments, which is capable of transducing non-neuronal cells, e.g., glial cells (e.g., oligodendrocytes or astrocytes).
  
  512. The AAV capsid variant of embodiment 511, wherein the non-neuronal cells comprise glial cells, oligodendrocytes (e.g., Olig2 positive oligodendrocytes), or astrocytes (e.g., Olig2 positive astrocytes).
  
  513. The AAV capsid variant of any one of the preceding embodiments, which is capable of transducing Olig2 positive cells, e.g., Olig2 positive astrocytes or Olig2 positive oligodendrocytes.
  
  514. The AAV capsid variant of any one of embodiments 369, 373, 447-454, 457, 458, or 482, which has increased tropism for a heart cell or tissue, e.g., a heart ventricle or heart atrium, relative to the tropism of a reference sequence of SEQ ID NO: 138.
  
  515. The AAV capsid variant of any one of embodiments 369, 373, 447-454, 457, 458, 482, or 514, which is enriched at least about 4, 5, 8, 10, 11, 12, 13, 14, 18, 19, 20, 21, 22, 24, 25, 27, 31, 33, or 34-fold, in the heart compared to a reference sequence of SEQ ID NO: 138, e.g., when measured by an assay as described in Example 4.
  
  516. The AAV capsid variant of any one of embodiments 369, 374, 447-454, 457, 458, 482, which has an increased tropism for a muscle cell or tissue (e.g., a quadriceps cell or a quadriceps tissue), relative to the tropism of a reference sequence comprising the amino acid sequence of SEQ ID NO: 138.
  
  517. The AAV capsid variant of any one of embodiments 369, 374, 447-454, 457, 458, 482, which is enriched at least about 4, 5, 8, 12, 17, 18, 20, 26, 27, 28, 30, or 36-fold, in the muscle compared to a reference sequence of SEQ ID NO: 138, e.g., when measured by an assay as described in Example 4.
  
  518. The AAV capsid variant of embodiment 516 or 517, wherein the muscle cell or tissue is a heart muscle (e.g., a heart ventricle or a heart atrium, or both), a quadriceps muscle, or both.
  
  519. The AAV capsid variant of any one of the preceding embodiments, which is isolated, e.g., recombinant.
  
  520. A polynucleotide encoding the AAV capsid variant of any one of embodiments 1-519.
  
  521. The polynucleotide of embodiment 520, which comprises:
- (i) a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequences of SEQ ID NO: 3 or 942;
- (ii) a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides, relative to the nucleotide sequences of SEQ ID NO: 3 or 942; or
- (iii) the nucleotide sequence of SEQ ID NOs: 3 or 942, or nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto.
  
  522. The polynucleotide of embodiment 520 or 521, which comprises the nucleotide sequence of SEQ ID NO: 983 or 984, or a nucleotide sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  523. The polynucleotide of any one of embodiments 520-522, which comprises a nucleotide sequence that is codon optimized.
  
  524. A peptide comprising an amino acid sequence having the following formula: [N1]-[N2]-[N3] according to any one of embodiments 1-8, 12-35, 39-45, 48-53, 91, or 118-123.
  
  525. A peptide comprising an amino acid sequence having the following formula: [A][B] according to any one of embodiments 130-133, 142, or 146.
  
  526. A peptide comprising an amino acid sequence having the following formula: [A][B] according to any one of embodiments 147-150, 166, or 167.
  
  527. A peptide comprising an amino acid sequence having the following formula: [N1]-[N2]-[N3] according to any one of embodiments 168-176, 179-186, 191-195, 198-202, or 295-299.
  
  528. A peptide comprising an amino acid sequence having the following formula: [N1]-[N2]-[N3] according to any one of embodiments 203-211, 214-223, 228-231, 234-239, or 295-299.
  
  529. A peptide comprising an amino acid sequence having the following formula: [A]-[B], according to any one of embodiments 300-306, 316, 317, 322, or 326.
  
  530. A peptide comprising:
- (a) the amino acid sequence of any of the sequences provided in Tables 1, 2A, 2B, 13-19;
- (b) an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 consecutive amino acids from any one of the sequences provided in Tables 1, 2A, 2B, 13-19;
- (c) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of any one of the sequences provided in Tables 1, 2A, 2B, 13-19; or
- (d) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of any one of the sequences provided in Tables 1, 2A, 2B, 13-19.
  
  531. A peptide comprising:
- (a) the amino acid sequence of any of SEQ ID NOs: 3849-4051 or 4681-4693;
- (b) an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 consecutive amino acids from any one of SEQ ID NOs: 3849-4051 or 4681-4693;
- (c) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of any one of SEQ ID NOs: 3849-4051 or 4681-4693; or
- (d) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of any one of SEQ ID NOs: 3849-4051 or 4681-4693.
  
  532. A peptide comprising:
- (a) the amino acid sequence of any of SEQ ID NOs: 4052-4092;
- (b) an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 consecutive amino acids from any one of SEQ ID NOs: 4052-4092;
- (c) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of any one of SEQ ID NOs: 4052-4092; or
- (d) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of any one of SEQ ID NOs: 4052-4092.
  
  533. A peptide comprising:
- (a) the amino acid sequence of any of SEQ ID NOs: 4056, 4058, 4059, 4062-4064, 4066, 4067, 4080, 4084, 4090, or 4095-4097;
- (b) an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 consecutive amino acids from any one of SEQ ID NOs: 4056, 4058, 4059, 4062-4064, 4066, 4067, 4080, 4084, 4090, or 4095-4097;
- (c) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of any one of SEQ ID NOs: 4056, 4058, 4059, 4062-4064, 4066, 4067, 4080, 4084, 4090, or 4095-4097; or
- (d) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of any one of SEQ ID NOs: 4056, 4058, 4059, 4062-4064, 4066, 4067, 4080, 4084, 4090, or 4095-4097.
  
  534. A peptide comprising:
- (a) the amino acid sequence of any of SEQ ID NOs: 945-980 or 985-986;
- (b) an amino acid sequence comprising at least 3, 4, or 5 consecutive amino acids from any one of SEQ ID NOs: 945-980 or 985-986;
- (c) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of any one of SEQ ID NOs: 945-980 or 985-986; or
- (d) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of any one of SEQ ID NOs: 945-980 or 985-986.
  
  535. A peptide comprising:
- (a) the amino acid sequence of any of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909;
- (b) an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 consecutive amino acids from any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909;
- (c) an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of any one of SEQ ID NOs: 2,200, 201, 941, 943, 204, 208, 404, or 903-909; or
- (d) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909.
  
  536. A peptide comprising:
- (i) the amino acid sequence of SPHSKA (SEQ ID NO: 941);
- (ii) an amino acid sequence comprising at least one, two, or three, but no more than four different amino acids relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941);
- (iii) an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941); or
- (iv) at least 3, 4, or 5 consecutive amino acids from the amino acid sequence of SPHSKA (SEQ ID NO: 941).
  
  537. A peptide encoded by:
- (i) the nucleotide sequence of SEQ ID NO: 942, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto;
- (ii) a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides relative to the nucleotide sequence of SEQ ID NO: 942; or
- (iii) a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of SEQ ID NO: 942.
  
  538. A peptide wherein the nucleotide sequence encoding the peptide comprises:
- (i) the nucleotide sequence of SEQ ID NO: 942, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto;
- (ii) a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but not more than 10 different nucleotides, relative to the nucleotide sequence of SEQ ID NO: 942; or
- (iii) a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of SEQ ID NO: 942.
  
  539. A peptide comprising:
- (i) the amino acid sequence of HDSPHK (SEQ ID NO: 2);
- (ii) an amino acid sequence comprising at least one, two, or three, but no more than four different amino acids, relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2);
- (iii) an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2); or
- (iv) at least 3, 4, or 5 consecutive amino acids from the amino acid sequence of HDSPHK (SEQ ID NO: 2).
  
  540. A peptide encoded by:
- (i) the nucleotide sequence of SEQ ID NO: 3, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or
- (ii) a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides, relative to the nucleotide sequence of SEQ ID NO: 3
- (iii) a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of SEQ ID NO: 3.
  
  541. A peptide wherein the nucleotide sequence encoding the peptide comprises:
- (i) the nucleotide sequence of SEQ ID NO: 3, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto;
- (ii) a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides relative to the nucleotide sequence of SEQ ID NO: 3; or
- (iii) a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of SEQ ID NO: 3.
  
  542. A peptide comprising:
- (i) the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589);
- (ii) an amino acid sequence comprising at least one, two, or three, but no more than four different amino acids, relative to the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589);
- (iii) an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589); or
- (iv) at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17 consecutive amino acids from the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589).
  
  543. A peptide comprising:
- (i) the amino acid sequence of KAEIGHDSPHKSGQNQQT (SEQ ID NO: 1754);
- (ii) an amino acid sequence comprising at least one, two, or three, but no more than four different amino acids, relative to the amino acid sequence of KAEIGHDSPHKSGQNQQT (SEQ ID NO: 1754);
- (iii) an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of KAEIGHDSPHKSGQNQQT (SEQ ID NO: 1754); or
- (iv) at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17 consecutive amino acids from the amino acid sequence of KAEIGHDSPHKSGQNQQT (SEQ ID NO: 1754).
  
  544. The peptide of any one of embodiments 524-543, which is fused or coupled, e.g., conjugated, to an active agent, e.g., a therapeutic agent or a diagnostic agent.
  
  545. The peptide of any one of embodiments 524-544, wherein at least 1-5, e.g., at least 1, 2, 3, 4, or 5, peptides are fused or coupled, e.g., conjugated, to an active agent, e.g., a therapeutic agent or a diagnostic agent.
  
  546. The peptide of embodiment 545, wherein the at least 1-5, e.g., at least 1, 2, 3, 4, or 5, peptides comprise the same amino acid sequence.
  
  547. The peptide of embodiment 454, wherein the at least 1-5, e.g., at least 1, 2, 3, 4, or 5, peptides comprise different amino acid sequences.
  
  548. The peptide of any one of embodiments 545-547, wherein the at least 1-5, e.g., at least 1, 2, 3, 4, or 5, peptides are present in tandem (e.g., connected directly or indirectly via a linker) or in a multimeric configuration.
  
  549. The peptide of any one of embodiments 524-548, wherein the peptide comprises an amino acid sequence of at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 20, 25, 30, or 35 amino acids in length.
  
  550. The peptide of any one of embodiments 544-549, wherein the active agent is or comprises a therapeutic agent chosen from a protein (e.g., an enzyme), an antibody molecule, a nucleic acid molecule (e.g., an RNAi agent), or a small molecule.
  
  551. The peptide of any one of embodiments 544-549, wherein the active agent is or comprises a ribonucleic acid complex (e.g., a Cas9/gRNA complex), a plasmid, a closed-end DNA, a circ-RNA, or an mRNA.
  
  552. The peptide of any one of embodiments 544-549, wherein the active agent is an RNAi agent.
  
  553. The peptide of embodiment 552, wherein the RNAi agent is a dsRNA, a siRNA, a shRNA, a pre-miRNA, a pri-miRNA, a miRNA, a stRNA, a lncRNA, a piRNA, an antisense oligonucleotide agent (ASO), or a snoRNA, optionally wherein the RNAi agent is an siRNA or an ASO, which further optionally comprises at least one modified nucleotide.
  
  554. The peptide of any one of embodiments 544-553, wherein the active agent modulates, e.g., inhibits, decreases or increases, expression of, a CNS related gene, mRNA, and/or protein.
  
  555. The peptide of any one of embodiments 544-449, wherein the active agent is a diagnostic agent is or comprises an imaging agent (e.g., a protein or small molecule compound coupled to a detectable moiety).
  
  556. The peptide of any one of embodiments 544-555, wherein the peptide covalently linked, e.g., directly or indirectly via a linker, to the active agent.
  
  557. The peptide of any one of embodiments 544-556, wherein the peptide is conjugated to the active agent via a linker.
  
  558. The peptide of embodiment 557, wherein the linker is a cleavable linker or a non-cleavable linker.
  
  559. The peptide of embodiment 558, wherein the cleavable linker is a pH sensitive linker or an enzyme sensitive linker.
  
  560. The peptide of embodiment 558 or 559, wherein:
- (i) the pH sensitive linker comprises a hydrazine/hydrazone linker or a disulfide linker;
- (ii) the enzyme sensitive linker comprises a peptide based linker, e.g., a peptide linker sensitive to a protease (e.g., a lysosomal protease); or a beta-glucuronide linker; or
- (iii) the non-cleavable linker is a linker comprising a thioether group or a maleimidocaproyl group.
  
  561. The peptide of any one of embodiments 544-560, wherein:
- (i) the peptide and the active agent are fused or coupled post-translationally, e.g., using click chemistry; or
- (ii) the peptide and the active agent are fused or couple via chemically induced dimerization.
  
  562. The peptide of any one of embodiments 544-561, wherein the peptide is present N-terminal relative to the active agent.
  
  563. The peptide of any one of embodiments 544-561, wherein the peptide is present C-terminal relative to the active agent.
  
  564. The peptide of any one of embodiment 544-549, 554, or 556-563, wherein the peptide is present or coupled to a carrier, e.g., an exosome, a microvesicle, or a lipid nanoparticle (LNP), optionally, wherein the carrier comprises a therapeutic agent (e.g., an RNAi agent (e.g., an dsRNA, a siRNA, a shRNA, a pre-miRNA, a pri-miRNA, a miRNA, a stRNA, a lncRNA, a piRNA, an antisense oligonucleotide agent (ASO), or a snoRNA), an mRNA, a ribonucleoprotein complex (e.g., a Cas9/gRNA complex), or a circRNA).
  
  565. The peptide of embodiment 564, wherein the peptide is present on the surface of the carrier, optionally wherein at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, or 80% of the surface of the carrier comprises at least 1-5, e.g., at least 1, 2, 3, 4, or 5 peptides according to any one of embodiments 422-436.
  
  566. An AAV capsid variant, comprising the peptide of any one of embodiments 524-543.
  
  567. A polynucleotide encoding an AAV capsid variant comprising:
- (a) the amino acid sequence of any of the sequences provided in Tables 1, 2A, 2B, 13-19;
- (b) an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 consecutive amino acids from any one of the sequences provided in Tables 1, 2A, 2B, 13-19;
- (c) an amino a sequence comprising at least one, two, or three but no more than four different amino acids, relative to the amino acid sequence of any one of the sequences provided in Tables 1, 2A, 2B, 13-19; or
- (d) an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (conservative substitutions), relative to the amino acid sequence of any one of the sequences provided in Tables 1, 2A, 2B, 13-19;
- optionally wherein the amino acid sequence of (a), (b), (c), and/or (d) is present immediately subsequent to position 448, 449, 450, 451, 452, 453, 454, or 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138, 981, or 982.
  
  568. A polynucleotide encoding an AAV capsid variant, wherein the AAV capsid variant comprises:
- (i) the amino acid sequence of SPHSKA (SEQ ID NO: 941);
- (ii) an amino acid sequence comprising at least one, two, or three, but no more than four different amino acids, relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941);
- (iii) an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941); or
- (iv) at least 3, 4, 5, 6, 7, 8, or 9 consecutive amino acids from the amino acid sequence of SPHSKA (SEQ ID NO: 941);
- optionally wherein the amino acid sequence of (i), (ii), (iii), and/or (iv) is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  569. The polynucleotide of embodiment 567 or 568, which comprises:
- (i) the nucleotide sequence of SEQ ID NO: 942, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto;
- (ii) a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of SEQ ID NO: 942;
- (iii) a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of SEQ ID NO: 942.
  
  570. A polynucleotide encoding an AAV capsid variant, wherein the AAV capsid variant comprises:
- (i) the amino acid sequence of HDSPHK (SEQ ID NO: 2);
- (ii) an amino acid sequence comprising at least one, two, or three, but no more than four different amino acids, relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2);
- (iii) an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2); or
- (iv) at least 3, 4, or 5 consecutive amino acids from the amino acid sequence of HDSPHK (SEQ ID NO: 2);
- optionally wherein the amino acid sequence of (i), (ii), (iii), and/or (iv) is present immediately subsequent to position 453, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.
  
  571. The polynucleotide of any one of embodiments 567 or 570, which comprises:
- (i) the nucleotide sequence of SEQ ID NO: 3, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto;
- (ii) a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides, relative to the nucleotide sequence of SEQ ID NO: 3; or
- (iii) a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of SEQ ID NO: 3.
  
  572. The polynucleotide of any one of embodiments 567-571, wherein the AAV capsid variant comprises:
- (i) the amino acid sequence of SEQ ID NO: 981 or 982, or an amino acid sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto;
- (ii) an amino acid sequence having at least one, two or three, but no more than four different amino acids, relative to the amino acid sequence of SEQ ID NO: 981 or 982; or
- (iii) an amino acid sequence having at least one, two or three modifications, e.g., substitutions (e.g., conservative substitutions), but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of SEQ ID NO: 981 or 982.
  
  573. The polynucleotide of any one of embodiments 567-572, comprising the nucleotide sequence of SEQ ID NO: 983 or 984, or a nucleotide sequence with at least 80% (e.g., at least about 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.
  
  574. The polynucleotide, peptide, or AAV capsid variant, of any one of embodiments 1-573, which is isolated, e.g., recombinant.
  
  575. An AAV particle comprising the AAV capsid variant of any one of embodiments 1-519, 566, or 574.
  
  576. The AAV particle of embodiment 575, which comprises a nucleotide sequence encoding a payload.
  
  577. The AAV particle of embodiment 576, wherein the encoded payload comprises a therapeutic protein or functional variant thereof; an antibody or antibody fragment; an enzyme; a component of a gene editing system; an RNAi agent (e.g., a dsRNA, siRNA, shRNA, pre-miRNA, pri-miRNA, miRNA, stRNA, lncRNA, piRNA, or snoRNA); or a combination thereof.
  
  578. The AAV particle of embodiment 577, wherein the therapeutic protein or functional variant thereof, e.g., a recombinant protein, is associated with (e.g., aberrantly expressed in) a neurological or neurodegenerative disorder, a muscular or neuromuscular disorder, or a neuro-oncological disorder.
  
  579. The AAV particle of embodiment 577 or 578, the therapeutic protein or functional variant thereof is chosen from apolipoprotein E (APOE) (e.g., ApoE2, ApoE3 and/or ApoE4); human survival of motor neuron (SMN) 1 or SMN2; glucocerebrosidase (GBA1); aromatic L-amino acid decarboxylase (AADC); aspartoacylase (ASPA); tripeptidyl peptidase I (CLN2); beta-galactosidase (GLB1); N-sulphoglucosamine sulphohydrolase (SGSH); N-acetyl-alpha-glucosaminidase (NAGLU); iduronate 2-sulfatase (IDS); intracellular cholesterol transporter (NPC1); gigaxonin (GAN); or a combination thereof.
  
  580. The AAV particle of embodiment 577, wherein the antibody or antibody binding fragment binds to:
- (i) a CNS related target, e.g., an antigen associated with a neurological or neurodegenerative disorder, e.g., β-amyloid, APOE, tau, SOD1, TDP-43, huntingtin (HTT), and/or synuclein;
- (ii) a muscular or neuromuscular related target, e.g., an antigen associated with a muscular or neuromuscular disorder; or
- (iii) a neuro-oncology related target, e.g., an antigen associated with a neuro-oncological disorder, e.g., HER2, or EGFR (e.g., EGFRvIII).
  
  581. The AAV particle of embodiment 577, wherein the enzyme comprises a meganuclease, a zinc finger nuclease, a TALEN, a recombinase, integrase, a base editor, a Cas9, or a fragment thereof.
  
  582. The AAV particle of embodiment 577, wherein the component of a gene editing system comprises one or more components of a CRISPR-Cas system.
  
  583. The AAV particle of embodiment 582, wherein the one or more components of the CRISPR-Cas system comprises a Cas9, e.g., a Cas9 ortholog or a Cpf1, and a single guide RNA (sgRNA), optionally wherein:
- (i) the sgRNA is located upstream (5′) of the cas9 enzyme; or
- (ii) the sgRNA is located downstream (3′) of the cas9 enzyme.
  
  584. The AAV particle of embodiment 577, wherein the RNAi agent (e.g., a dsRNA, siRNA, shRNA, pre-miRNA, pri-miRNA, miRNA, stRNA, lncRNA, piRNA, or snoRNA), modulates, e.g., inhibits, expression of, a CNS related gene, mRNA, and/or protein.
  
  585. The AAV particle of embodiment 584, wherein the CNS related gene is chosen from SOD1, MAPT, APOE, HTT, C90RF72, TDP-43, APP, BACE, SNCA, ATXN1, ATXN3, ATXN7, SCN1A-SCN5A, SCN8A-SCN11A, or a combination thereof.
  
  586. The AAV particle of any one of embodiments 575-585, which comprises a viral genome comprising a promoter operably linked to the nucleic acid sequence encoding the payload.
  
  587. The AAV particle of embodiment 586, wherein the promoter is chosen from human elongation factor 1α-subunit (EF1α), cytomegalovirus (CMV) immediate-early enhancer and/or promoter, chicken β-actin (CBA) and its derivative CAG, β glucuronidase (GUSB), or ubiquitin C (UBC), neuron-specific enolase (NSE), platelet-derived growth factor (PDGF), platelet-derived growth factor B-chain (PDGF-β), intercellular adhesion molecule 2 (ICAM-2), synapsin (Syn), methyl-CpG binding protein 2 (MeCP2), Ca2+/calmodulin-dependent protein kinase II (CaMKII), metabotropic glutamate receptor 2 (mGluR2), neurofilament light (NFL) or heavy (NFH), β-globin minigene nβ2, preproenkephalin (PPE), enkephalin (Enk) and excitatory amino acid transporter 2 (EAAT2), glial fibrillary acidic protein (GFAP), myelin basic protein (MBP), a cardiovascular promoter (e.g., αMHC, cTnT, and CMV-MLC2k), a liver promoter (e.g., hAAT, TBG), a skeletal muscle promoter (e.g., desmin, MCK, C512) or a fragment, e.g., a truncation, or a functional variant thereof.
  
  588. The AAV particle of embodiment 586 or 587, wherein the promoter is an EF-1a promoter variant, e.g., a truncated EF-1a promoter.
  
  589. The AAV particle of any one of embodiments 586-588, wherein the promoter comprises the nucleotide sequence of any one of SEQ ID NOs: 987, 988, 990, 991, 995, 996, 998-1007 or any one of the sequences provided in Table 8, a nucleotide sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of SEQ ID NOs: 987, 988, 990, 991, 995, 996, 998-1007 or any one of the sequences provided in Table 8, or a nucleotide sequence with at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to any one of SEQ ID NOs: 987, 988, 990, 991, 995, 996, 998-1007 or any one of the sequences provided in Table 8.
  
  590. The AAV particle of any one of embodiments 586-589 wherein the viral genome further comprises a polyA signal sequence.
  
  591. The AAV particle of any one of embodiments 586-590, wherein the viral genome further comprises an inverted terminal repeat (ITR) sequence.
  
  592. The AAV particle of any one of embodiments 586-591, wherein the viral genome comprises an ITR sequence positioned 5′ relative to the encoded payload.
  
  593. The AAV particle of any one of embodiments 586-592, wherein the viral genome comprises an ITR sequence positioned 3′ relative to the encoded payload.
  
  594. The AAV particle of any one of embodiments 586-593, wherein the viral genome comprises an ITR sequence positioned 5′ relative to the encoded payload and an ITR sequence positioned 3′ relative to the encoded payload.
  
  595. The AAV particle of any one of embodiments 586-594, wherein the viral genome further comprises an enhancer, a Kozak sequence, an intron region, and/or an exon region.
  
  596. The AAV particle of any one of embodiments 586-594, wherein the viral genome further comprises a nucleotide sequence encoding a miR binding site, e.g., a miR binding site that modulates, e.g., reduces, expression of the antibody molecule encoded by the viral genome in a cell or tissue where the corresponding miRNA is expressed.
  
  597. The AAV particle of embodiment 596, wherein the encoded miRNA binding site is complementary, e.g., fully complementary or partially complementary, to a miRNA expressed in a cell or tissue of the DRG, liver, heart, hematopoietic, or a combination thereof.
  
  598. The AAV particle of embodiment 596 or 597, wherein the encoded miR binding site modulates, e.g., reduces, expression of the encoded antibody molecule in a cell or tissue of the DRG, liver, heart, hematopoietic lineage, or a combination thereof.
  
  599. The AAV particle of any one of embodiments 586-598, wherein the viral genome comprises at least 1-5 copies of the encoded miR binding site, e.g., at least 1, 2, 3, 4, or 5 copies.
  
  600. The AAV particle of any one of embodiments 586-599, wherein the viral genome comprises at least 3 copies of an encoded miR binding sites, optionally wherein all three copies comprise the same miR binding site, or at least one, two, three, or all of the copies comprise a different miR binding site.
  
  601. The AAV particle of embodiment 600, wherein the 3 copies of the encoded miR binding sites are continuous (e.g., not separated by a spacer), or are separated by a spacer, optionally wherein the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of GATAGTTA.
  
  602. The AAV particle of any one of embodiments 586-601, wherein the viral genome comprises at least 4 copies of an encoded miR binding site, optionally wherein all four copies comprise the same miR binding site, or at least one, two, three, or all of the copies comprise a different miR binding site.
  
  603. The AAV particle of embodiment 602, wherein the 4 copies of the encoded miR binding sites are continuous (e.g., not separated by a spacer), or are separated by a spacer, optionally wherein the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of GATAGTTA.
  
  604. The AAV particle of any one of embodiments 596-603, wherein the encoded miR binding site comprises a miR122 binding site, a miR183 binding site, a miR-1 binding site, a miR-142-3p, or a combination thereof, optionally wherein:
- (i) the encoded miR122 binding site comprises the nucleotide sequence of SEQ ID NO: 4673, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4673;
- (ii) the encoded miR183 binding site comprises the nucleotide sequence of SEQ ID NO: 4676, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4676;
- (iii) the encoded miR-1 binding site comprises the nucleotide sequence of SEQ ID NO: 4679, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4679; and/or
- (iv) the encoded miR-142-3p binding site comprises the nucleotide sequence of SEQ ID NO: 4675, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4675.
  
  605. The AAV particle of any one of embodiments 586-604, wherein the viral genome comprises an encoded miR122 binding site.
  
  606. The AAV particle of any one of embodiments 586-605, wherein the viral genome comprises at least 1-5 copies, e.g., 1, 2, or 3 copies of a miR122 binding site, optionally wherein each copy is continuous (e.g., not separated by a spacer), or each copy is separated by a spacer, optionally wherein the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of GATAGTTA.
  
  607. The AAV particle of embodiment 605 or 606, wherein the encoded miR122 binding site comprises the nucleotide sequence of SEQ ID NO: 4673, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4673.
  
  608. The AAV particle of any one of embodiments 586-607, wherein the viral genome comprises:
- (A) (i) a first encoded miR122 binding site comprising the nucleotide sequence of SEQ ID NO: 4673, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4673;
  - (ii) a first spacer comprising the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of GATAGTTA; and
  - (iii) a second encoded miR122 binding site comprising the nucleotide sequence of SEQ ID NO: 4673, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4673; or
- (B) (i) a first encoded miR122 binding site comprising the nucleotide sequence of SEQ ID NO: 4673, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4673;
  - (ii) a first spacer comprising the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of GATAGTTA;
  - (iii) a second encoded miR122 binding site comprising the nucleotide sequence of SEQ ID NO: 4673, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4673;
  - (iv) a second spacer comprising the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of GATAGTTA; and
  - (v) a third encoded miR122 binding site comprising the nucleotide sequence of SEQ ID NO: 4673, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4673.
    
    609. The AAV particle of any one of embodiments 586-608, wherein the viral genome comprises an encoded miR183 binding site.
    
    610. The AAV particle of any one of embodiments 586-609, wherein the viral genome comprises at least 1-5 copies, e.g., 1, 2, or 3 copies of a miR183 binding site, optionally wherein each copy is continuous (e.g., not separated by a spacer), or each copy is separated by a spacer, optionally wherein the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of GATAGTTA.
    
    611. The AAV particle of embodiment 609 or 610, wherein the encoded miR183 binding site comprises the nucleotide sequence of SEQ ID NO: 4673, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4673.
    
    612. The AAV particle of any one of embodiments 586-611, wherein the viral genome comprises:
- (A) (i) a first encoded miR183 binding site comprising the nucleotide sequence of SEQ ID NO: 4676, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4676;
  - (ii) a first spacer comprising the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of GATAGTTA; and
  - (iii) a second encoded miR183 binding site comprising the nucleotide sequence of SEQ ID NO: 4676, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4676; or
- (B) (i) a first encoded miR183 binding site comprising the nucleotide sequence of SEQ ID NO: 4676, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4676;
  - (ii) a first spacer comprising the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to GATAGTTA;
  - (iii) a second encoded miR183 binding site comprising the nucleotide sequence of SEQ ID NO: 4676, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4676;
  - (iv) a second spacer comprising the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of GATAGTTA; and
  - (v) a third encoded miR183 binding site comprising the nucleotide sequence of SEQ ID NO: 4676, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto; or a nucleotide sequence having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), relative to SEQ ID NO: 4676.
    
    613. The AAV particle of any one of embodiments 586-612, wherein the viral genome comprises an encoded miR122 binding site and a miR-1 binding site.
    
    614. The AAV particle of any one of embodiments 586-613, wherein the viral genome is single stranded.
    
    615. The AAV particle of any one of embodiments 586-613, wherein the viral genome self-complementary.
    
    616. The AAV particle of any one of embodiments 586-615, wherein the viral genome further comprises a nucleotide sequence encoding a Rep protein, e.g., a non-structural protein, wherein the Rep protein comprises a Rep78 protein, a Rep68, Rep52 protein, and/or a Rep40 protein (e.g., a Rep78 and a Rep52 protein).
    
    617. The AAV particle of any one of embodiments 586-615, wherein the AAV particle further comprises a nucleotide sequence encoding a Rep protein, e.g., a non-structural protein, wherein the Rep protein comprises a Rep78 protein, a Rep68, Rep52 protein, and/or a Rep40 protein (e.g., a Rep78 and a Rep52 protein).
    
    618. The AAV particle of embodiment 616 or 617, wherein the Rep78 protein, the Rep68 protein, the Rep52 protein, and/or the Rep40 protein are encoded by at least one Rep gene.
    
    619. The AAV particle of any one of embodiments 586-618, wherein the viral genome further comprises a nucleic acid sequence encoding the AAV capsid variant of any one of embodiments 1-519, 566, or 574.
    
    620. The AAV particle of any one of embodiments 575-519, which is isolated, e.g., recombinant.
    
    621. A vector comprising a polynucleotide encoding the AAV capsid variant of any one of embodiments 1-519, 566, or 574, the polynucleotide of any one of embodiments 520-523 or 567-574, or a polynucleotide encoding the peptide of any one of embodiments 524-543 or 574.
    
    622. A cell, e.g., a host cell, comprising the AAV capsid variant of any one of embodiments 1-519, 566, or 574, the polynucleotide of any one of embodiments 520-523 or 567-574, the peptide of any one of embodiments 524-565 or 574, the AAV particle of any one of embodiments 575-620, or the vector of embodiment 621.
    
    623. The cell of embodiment 622, wherein the cell is a mammalian cell or an insect cell.
    
    624. The cell of embodiment 622 or 623, wherein the cell is a cell of a brain region or a spinal cord region, optionally a cell of the brain stem, hippocampus, or thalamus.
    
    625. The cell of any one of embodiments 622-624, wherein the cell is a neuron, a sensory neuron, a motor neuron, an astrocyte, a glial cell, oligodendrocyte, or a muscle cell (e.g., a cell of the heart, diaphragm, or quadriceps).
    
    626. The cell of embodiment 622 or 623, wherein the cell is a liver cell.
    
    627. A method of making an AAV particle, comprising
- (i) providing a host cell comprising a viral genome; and
- (ii) incubating the host cell under conditions suitable to enclose the viral genome in the AAV capsid variant of any one of embodiments 1-519, 566, or 574 or an AAV capsid variant encoded by the polynucleotide of any one of embodiments 520-523 or 567-574;
- thereby making the AAV particle.
  
  628. The method of embodiment 627, further comprising, prior to step (i), introducing a first nucleic acid molecule comprising the viral genome into the host cell.
  
  629. The method of embodiment 628, wherein the host cell comprises a second nucleic acid encoding the capsid variant.
  
  630. The method of embodiment 629, wherein the second nucleic acid molecule is introduced into the host cell prior to, concurrently with, or after the first nucleic acid molecule.
  
  631. A pharmaceutical composition comprising the AAV particle of any one of embodiments 575-620, an AAV particle comprising the capsid variant of any one of embodiments 1-519, 566, or 574, an AAV particle comprising the peptide of any one of embodiments 524-543 or 574, and a pharmaceutically acceptable excipient.
  
  632. A method of delivering a payload to a cell or tissue (e.g., a CNS cell, CNS tissue, a liver cell, or a liver tissue), comprising administering an effective amount of the pharmaceutical composition of embodiment 631, the AAV particle of any one of embodiments 575-620, an AAV particle comprising the capsid variant of any one of embodiments 1-519, 566, or 574, or an AAV particle comprising the peptide of any one of embodiments 524-543 or 574.
  
  633. The method of embodiment 632, wherein the cell is a cell of a brain region or a spinal cord region, optionally a cell of the frontal cortex, sensory cortex, motor cortex, caudate, cerebellar cortex, cerebral cortex, brain stem, hippocampus, or thalamus.
  
  634. The method of embodiment 632 or 633, wherein the cell is a neuron, a sensory neuron, a motor neuron, an astrocyte, a glial cell, or an oligodendrocyte.
  
  635. The method of embodiment 632, wherein the cell is a liver cell.
  
  636. The method of any one of embodiments 632-635, wherein the cell or tissue is within a subject.
  
  637. The method of embodiment 636, wherein the subject has, has been diagnosed with having, or is at risk of having a genetic disorder, e.g., a monogenic disorder or a polygenic disorder.
  
  638. The method of embodiment 636 or 637, wherein the subject has, has been diagnosed with having, or is at risk of having a neurological, e.g., a neurodegenerative disorder.
  
  639. The method of embodiment 636 or 637, wherein the subject has, has been diagnosed with having, or is at risk of having a neuro-oncological disorder.
  
  640. The method of embodiment 636 or 637, wherein the subject has, has been diagnosed with having, or is at risk of having a muscular disorder or a neuromuscular disorder.
  
  641. A method of treating a subject having or diagnosed with having a genetic disorder, e.g., a monogenic disorder or a polygenic disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of embodiment 631, the AAV particle of any one of embodiments 575-620, an AAV particle comprising the capsid variant of any one of embodiments 1-519, 566, or 574, or an AAV particle comprising the peptide of any one of embodiments 524-543 or 574.
  
  642. A method of treating a subject having or diagnosed with having a neurological disorder, e.g., a neurodegenerative disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of embodiment 631, the AAV particle of any one of embodiments 575-620, an AAV particle comprising the capsid variant of any one of embodiments 1-519, 566, or 574, or an AAV particle comprising the peptide of any one of embodiments 524-543 or 574.
  
  643. A method of treating a subject having or diagnosed with having a muscular disorder or a neuromuscular disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of embodiment 631, the AAV particle of any one of embodiments 575-620, an AAV particle comprising the capsid variant of any one of embodiments 1-519, 566, or 574, or an AAV particle comprising the peptide of any one of embodiments 524-543 or 574.
  
  644. A method of treating a subject having or diagnosed with having a neuro-oncological disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of embodiment 631, the AAV particle of any one of embodiments 575-620, an AAV particle comprising the capsid variant of any one of embodiments 1-519, 566, or 574, or an AAV particle comprising the peptide of any one of embodiments 524-543 or 574.
  
  645. The method of any one of embodiments 637-645, wherein the genetic disorder, neurological disorder, neurodegenerative disorder, muscular disorder, neuromuscular disorder, or neuro-oncological disorder is Huntington's Disease, Amyotrophic Lateral Sclerosis (ALS), Gaucher Disease, Dementia with Lewy Bodies, Parkinson's disease, Spinal Muscular Atrophy, Alzheimer's Disease, a leukodystrophy (e.g., Alexander disease, autosomal dominant leukodystrophy with autonomic diseases (ADLD), Canavan disease, cerebrotendinous xanthomatosis (CTX), metachromatic leukodystrophy (MLD), Pelizaeus-Merzbacher disease, or Refsum disease), or a cancer (e.g., a HER2/neu positive cancer or a glioblastoma)′.
  
  646. The method of any one of embodiments 641-645, where treating comprises prevention of progression of the disease or disorder in the subject.
  
  647. The method of embodiment 636-646, wherein the subject is a human.
  
  648. The method of any one of embodiments 636-647, wherein the AAV particle is administered to the subject intravenously, via intra-cisterna magna injection (ICM), intracerebrally, intrathecally, intracerebroventricularly, via intraparenchymal administration, intraarterially, or intramuscularly.
  
  649. The method of any one of embodiments 636-648, wherein the AAV particle is administered to the subject via focused ultrasound (FUS), e.g., coupled with the intravenous administration of microbubbles (FUS-MB), or MRI-guided FUS coupled with intravenous administration.
  
  650. The method of any one of embodiments 636-649, wherein the AAV particle is administered to the subject intravenously.
  
  651. The method of any one of embodiments 636-650, wherein the AAV particle is administered to the subject via intra-cisterna magna injection (ICM).
  
  652. The method of any one of embodiments 636-651, wherein the AAV particle is administered to the subject intraarterially.
  
  653. The method of any one of embodiments 648-652, wherein administration of the AAV particle results in a decreased presence, level, and/or activity of a gene, mRNA, protein, or combination thereof.
  
  654. The method of any one of embodiments 648-652, wherein administration of the AAV particle results in an increased presence, level, and/or activity of a gene, mRNA, protein, or a combination thereof.
  
  655. The pharmaceutical composition of embodiment 631, the AAV particle of any one of embodiments 575-620, an AAV particle comprising the capsid variant of any one of embodiments 1-519, 566, or 574, or an AAV particle comprising the peptide of any one of embodiments 524-543 or 574, for use in a method of delivering a payload to a cell or tissue.
  
  656. The pharmaceutical composition of embodiment 631, the AAV particle of any one of embodiments 575-620, an AAV particle comprising the capsid variant of any one of embodiments 1-519, 566, or 574, or an AAV particle comprising the peptide of any one of embodiments 524-543 or 574, for use in a method of treating a genetic disorder, a neurological disorder, a neurodegenerative disorder, a muscular disorder, a neuromuscular disorder, or a neuro-oncological disorder.
  
  657. The pharmaceutical composition of embodiment 631, the AAV particle of any one of embodiments 575-620, an AAV particle comprising the capsid variant of any one of embodiments 1-519, 566, or 574, or an AAV particle comprising the peptide of any one of embodiments 524-543 or 574, for use in the manufacture of a medicament.
  
  658. Use of the pharmaceutical composition of embodiment 631, the AAV particle of any one of embodiments 575-620, an AAV particle comprising the capsid variant of any one of embodiments 1-519, 566, or 574, or an AAV particle comprising the peptide of any one of embodiments 524-543 or 574, in the manufacture of a medicament.
  
  659. Use of the pharmaceutical composition of embodiment 631, the AAV particle of any one of embodiments 575-620, an AAV particle comprising the capsid variant of any one of embodiments 1-519, 566, or 574, or an AAV particle comprising the peptide of any one of embodiments 524-543 or 574, in the manufacture of a medicament for treating a genetic disorder, a neurological disorder, a neurodegenerative disorder, a muscular disorder, a neuromuscular disorder, or a neuro-oncological disorder.

The details of one or more embodiments of the disclosure are set forth in the accompanying description below. Other features, objects and advantages of the disclosure will be apparent from the description. In the description, the singular forms also include the plural unless the context clearly dictates otherwise. Certain terms are defined in the Definition section and throughout.

DETAILED DESCRIPTION OF THE DISCLOSURE

Described herein, inter alia, are compositions comprising an AAV capsid variant, e.g., an AAV capsid variant described herein, and methods of making and using the same. Generally, the AAV capsid variant has enhanced tropism for a cell or tissue, e.g., for the delivery of a payload to said cell or tissue, for example a CNS tissue or a CNS cell or a liver cell or liver tissue.

As demonstrated in the Examples herein below, certain AAV capsid variants described herein show multiple advantages over wild-type AAV9, including (i) increased penetrance through the blood brain barrier following intravenous administration, (ii) wider distribution throughout the multiple brain regions, e.g., frontal cortex, sensory cortex, motor cortex, putamen, thalamus, cerebellar cortex, dentate nucleus, caudate, and/or hippocampus, and/or (iii) elevated payload expression in multiple brain regions. Without wishing to be being bound by theory, it is believed that these advantages may be due, in part, to the dissemination of the AAV capsid variants through the brain vasculature. In some embodiments, the AAV capsids described herein enhance the delivery of a payload to multiple regions of the brain including for example, the frontal cortex, sensory cortex, motor cortex, putamen, thalamus, cerebellar cortex, dentate nucleus, caudate, and/or hippocampus.

Several approaches have been used previously to produce AAV capsids with enhanced tropism for a cell or tissue, e.g., a CNS cell or tissue. One approach used co-infection of cultured cells (Grimm et al. In vitro and in vivo gene therapy vector evolution via multispecies interbreeding and retargeting of adeno-associated viruses. J. Virol. 2008 June 82(12):5887-5911, the contents of which are herein incorporated by reference in its entirety) or in situ animal tissue (Lisowski et al. Selection and evaluation of clinically relevant AAV variants in a xenograft liver model. Nature 2014 506:382-386, the contents of which are herein incorporated by reference in its entirety) with adenovirus, in order to trigger exponential replication of infectious AAV DNA. Another approach involved the use of cell-specific CRE transgenic mice (Deverman et al. Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain. Nat Biotechnol. 2016 Feb. 34(2)204-209; the contents of which are herein incorporated by reference in its entirety) allowing viral DNA recombination specifically in astrocytes, followed by recovery of CRE-recombined capsid variants. Other approaches apply high throughput DNA synthesis, multiplexing, sequencing technologies, and machine learning to evaluate sequencing reads of viral DNA in different tissues to engineer variant capsids. These approaches are different from the approach disclosed herein.

There are some limitations to the art-known capsid generation methods. For example, the transgenic CRE system used by Deverman et al. (2016) has limited tractable in other animal species and AAV variants selected by directed evolution in mouse tissue do not show similar properties in large animals. Previously described transduction-specific approaches are not amenable to large animal studies because: 1) many tissues of interest (e.g., CNS) are not readily accessible to adenovirus co-infection, 2) the specific adenovirus tropism itself would bias the library distribution, and 3) large animals are typically not amenable to transgenesis or genetic engineering to express CRE recombinase in defined cell types.

To address these limitations, a broadly-applicable functional AAV capsid library screening platform for cell type-specific biopanning in non-transgenic animals has been developed and is described in the appended Examples. In the TRACER (Tropism Redirection of AAV by Cell type-specific Expression of RNA) platform system, the capsid gene is placed under the control of a cell type-specific promoter to drive capsid mRNA expression in the absence of helper virus co-infection. Without wishing to be bound by theory, it is believed that this RNA-driven screen increases the selective pressure in favor of capsid variants which transduce a specific cell type. The TRACER platform allows for generation of AAV capsid libraries whereby specific recovery and subcloning of capsid mRNA expressed in transduced cells is achieved with no need for transgenic animals or helper virus co-infection. Without wishing to be bound by theory, it is believed that since mRNA transcription is a hallmark of full transduction, the methods disclosed herein allow identification of fully infectious AAV capsid mutants, and in addition to its higher stringency, this method allows identification of capsids with high tropism for particular cell types using libraries designed to express CAP mRNA under the control of any cell-specific promoter such as, but not limited to, synapsin-1 promoter (neurons), GFAP promoter (astrocytes), TBG promoter (liver), CAMK promoter (skeletal muscle), MYH6 promoter (cardiomyocytes). Described herein are novel AAV capsid variants generated using the TRACER method which demonstrate enhance tropism in for example a CNS cell, a CNS tissue, a liver cell, a liver tissue, a muscle cell, or a muscle tissue.

In some embodiments, an AAV capsid variant disclosed herein comprises a modification in loop IV of AAV9, e.g., at positions between 449-460, e.g., at position 454 and/or 455, numbered relative to SEQ ID NO: 138, 981, or 982. In some embodiments, loop (e.g., loop IV) is used interchangeably herein with the term variable region (e.g., variable region IV), or VR (e.g., VR-IV). In some embodiments loop IV comprises positions 449-475 (e.g., amino acids KTINGSGQNQQTLKFSVAGPSNMAVQG (SEQ ID NO: 6404)), numbered according to SEQ ID NO: 138. In some embodiments loop IV comprises positions 449-460 (e.g., amino acids KTINGSGQNQQT (SEQ ID NO: 6405)), numbered according to SEQ ID NO: 138. In some embodiments, loop IV or variable region IV (VR-IV) is as described in DiMattia et al. “Structural Insights into the Unique Properties of the Adeno-Associated Virus Serotype 9,” Journal of Virology, 12(86):6947-6958 (the contents of which are hereby incorporated by reference in their entirety), e.g., comprising positions 452-460 (e.g., NGSGQNQQT (SEQ ID NO: 4487)), numbered according to SEQ ID NO: 138.

The AAV particles and payloads of the disclosure may be delivered to one or more target cells, tissues, organs, or organisms. In some embodiments, the AAV particles of the disclosure demonstrate enhanced tropism for a target cell type, tissue or organ. As a non-limiting example, the AAV particle may have enhanced tropism for cells and tissues of the central or peripheral nervous systems (CNS and PNS, respectively). In some embodiments, an AAV particle of the disclosure may, in addition, or alternatively, have decreased tropism for a cell-type, tissue or organ.

In some embodiments, an AAV comprises a small non-enveloped icosahedral capsid virus of the Parvoviridae family and is characterized by a single stranded DNA viral genome. Parvoviridae family viruses consist of two subfamilies: Parvovirinae, which infect vertebrates, and Densovirinae, which infect invertebrates. The Parvoviridae family comprises the Dependovirus genus which includes AAV, capable of replication in vertebrate hosts including, but not limited to, human, primate, bovine, canine, equine, and ovine species.

The parvoviruses and other members of the Parvoviridae family are generally described in Kenneth I. Berns, “Parvoviridae: The Viruses and Their Replication,” Chapter 69 in FIELDS VIROLOGY (3d Ed. 1996), the contents of which are incorporated by reference in their entirety.

In some embodiments, AAV are used as a biological tool due to a relatively simple structure, their ability to infect a wide range of cells (including quiescent and dividing cells) without integration into the host genome and without replicating, and their relatively benign immunogenic profile. The genome of the virus may be manipulated to contain a minimum of components for the assembly of a functional recombinant virus, or viral particle, which is loaded with or engineered to target a particular tissue and express or deliver a desired payload.

In some embodiments, the AAV, is a naturally occurring (e.g., wild-type) AAV or a recombinant AAV. In some embodiments, the wild-type AAV vector genome is a linear, single-stranded DNA (ssDNA) molecule approximately 5,000 nucleotides (nt) in length. In some embodiments, inverted terminal repeats (ITRs) cap the viral genome at both the 5′ and the 3′ end, providing origins of replication for the viral genome. In some embodiments, an AAV viral genome typically comprises two ITR sequences. These ITRs have a characteristic T-shaped hairpin structure defined by a self-complementary region (145nt in wild-type AAV) at the 5′ and 3′ ends of the ssDNA which form an energetically stable double stranded region. The double stranded hairpin structures comprise multiple functions including, but not limited to, acting as an origin for DNA replication by functioning as primers for the endogenous DNA polymerase complex of the host viral replication cell.

In some embodiments, the wild-type AAV viral genome further comprises nucleotide sequences for two open reading frames, one for the four non-structural Rep proteins (Rep78, Rep68, Rep52, Rep40, encoded by Rep genes) and one for the three capsid, or structural, proteins (VP1, VP2, VP3, encoded by capsid genes or Cap genes). The Rep proteins are used for replication and packaging, while the capsid proteins are assembled to create the protein shell of the AAV, or AAV capsid polypeptide, e.g., an AAV capsid variant. Alternative splicing and alternate initiation codons and promoters result in the generation of four different Rep proteins from a single open reading frame and the generation of three capsid proteins from a single open reading frame. Though it varies by AAV serotype, as a non-limiting example, for AAV9/hu.14 (SEQ ID NO: 123 of U.S. Pat. No. 7,906,111, the contents of which are herein incorporated by reference in their entirety) VP1 refers to amino acids 1-736, VP2 refers to amino acids 138-736, and VP3 refers to amino acids 203-736. In some embodiments, for any one of the amino acid sequences of SEQ ID NO: 981 or 982, VP1 comprises amino acids 1-742, VP2 comprises amino acids 138-742, and VP3 comprises amino acids 203-742. In other words, VP1 is the full-length capsid sequence, while VP2 and VP3 are shorter components of the whole. As a result, changes in the sequence in the VP3 region, are also changes to VP1 and VP2, however, the percent difference as compared to the parent sequence will be greatest for VP3 since it is the shortest sequence of the three. Though described here in relation to the amino acid sequence, the nucleic acid sequence encoding these proteins can be similarly described. Together, the three capsid proteins assemble to create the AAV capsid protein. While not wishing to be bound by theory, the AAV capsid protein typically comprises a molar ratio of 1:1:10 of VP1:VP2:VP3.

AAV vectors of the present disclosure may be produced recombinantly and may be based on adeno-associated virus (AAV) reference sequences. In addition to single stranded AAV viral genomes (e.g., ssAAVs), the present disclosure also provides for self-complementary AAV (scAAVs) viral genomes. scAAV vector genomes contain DNA strands which anneal together to form double stranded DNA. By skipping second strand synthesis, scAAVs allow for rapid expression in the transduced cell. In some embodiments, the AAV particle of the present disclosure is an scAAV. In some embodiments, the AAV particle of the present disclosure is an ssAAV.

Methods for producing and/or modifying AAV particles are disclosed in the art such as pseudotyped AAV vectors (PCT Patent Publication Nos. WO200028004; WO200123001; WO2004112727; WO2005005610; and WO2005072364, the content of each of which is incorporated herein by reference in its entirety).

As described herein, the AAV particles of the disclosure comprising an AAV capsid variant, and a viral genome, have enhanced tropism for a cell-type or a tissue, e.g., a CNS cell-type, region, or tissue.

Peptides

Disclosed herein are peptides, and associated AAV particles comprising an AAV capsid variant and a peptide for enhanced or improved transduction of a target tissue (e.g., cells of the CNS or PNS). In some, embodiments, the peptide is an isolated, e.g., recombinant, peptide. In some embodiments, the nucleic acid encoding the peptide, is an isolated, e.g., recombinant nucleic acid.

In some embodiments, the peptide may increase distribution of an AAV particle to a cell, region, or tissue of the CNS. The cell of the CNS may be, but is not limited to, neurons (e.g., excitatory, inhibitory, motor, sensory, autonomic, sympathetic, parasympathetic, Purkinje, Betz, etc.), glial cells (e.g., microglia, astrocytes, oligodendrocytes) and/or supporting cells of the brain such as immune cells (e.g., T cells). The tissue of the CNS may be, but is not limited to, the cortex (e.g., frontal, parietal, occipital, and/or temporal), thalamus, hypothalamus, striatum, putamen, caudate nucleus, hippocampus, entorhinal cortex, basal ganglia, or deep cerebellar nuclei.

In some embodiments, the peptide may increase distribution of an AAV particle to a cell, region, or tissue of the PNS. The cell or tissue of the PNS may be, but is not limited to, a dorsal root ganglion (DRG).

In some embodiments, the peptide may increase distribution of an AAV particle to the CNS (e.g., the cortex) after intravenous administration. In some embodiments, the peptide may increase distribution of an AAV particle to the CNS (e.g., the cortex) following focused ultrasound (FUS), e.g., coupled with the intravenous administration of microbubbles (FUS-MB), or MRI-guided FUS coupled with intravenous administration.

In some embodiments, the peptide may increase distribution of an AAV particle to the PNS (e.g., DRG) after intravenous administration. In some embodiments, the peptide may increase distribution of an AAV particle to the PNS (e.g., DRG) following focused ultrasound (FUS), e.g., coupled with the intravenous administration of microbubbles (FUS-MB), or MRI-guided FUS coupled with intravenous administration.

In some embodiments, the peptide may increase distribution of an AAV particle to a cell, region, or tissue of a muscle. In some embodiments, the muscle is a heart muscle, e.g., a heart atrium or a heart ventricle. In some embodiments, the peptide may direct an AAV particle to a muscle cell, region, or tissue after intravenous administration.

In some embodiments, the peptide may increase distribution of an AAV particle to a cell, region, or tissue of the liver.

A peptide may vary in length. In some embodiments, the peptide is about 3 to about 20 amino acids in length. As non-limiting examples, the peptide may be 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 3-5, 3-8, 3-10, 3-12, 3-15, 3-18, 3-20, 5-10, 5-15, 5-20, 10-12, 10-15, 10-20, 12-20, or 15-20 amino acids in length. In some embodiments, a peptide comprises about 6 to 12 amino acids in length, e.g., about 9 amino acids in length. In some embodiments, a peptide comprises about 5 to 10 amino acids in length, e.g., about 7 amino acids in length. In some embodiments, a peptide comprises about 7 to 11 amino acids in length, e.g., about 8 amino acids in length. In some embodiments, a peptide comprises about 4 to 9 amino acids in length, e.g., about 6 amino acids in length.

In some embodiments a peptide may comprise a sequence as set forth in Table 1 (e.g., comprising the amino acid sequence of any one of SEQ ID NOs: 200-940, 1800-2241, 2242-2886, or 2887-3076). In some embodiments a peptide may comprise a sequence as set forth in Table 2A or 2B. In some embodiments, the peptide may comprise a sequence set forth in Table 13 or 14. In some embodiments, the peptide may comprise a sequence as set forth in Table 15. In some embodiments, the peptide may comprise a sequence as set forth in Table 16. In some embodiments, the peptide may comprise a sequence as set forth in Table 17. In some embodiments, the peptide may comprise a sequence as set forth in Table 18. In some embodiments, the peptide may comprise a sequence as set forth in Table 19. In some embodiments, the peptide is isolated, e.g., recombinant.

TABLE 1

Exemplary Peptide Sequences

SEQ

SEQ

SEQ

SEQ

ID

ID
Peptide
ID
Peptide
ID

Peptide Sequence
NO:
Peptide Sequence
NO:
Sequence
NO:
Sequence
NO:

GSGSPHSKAQN
200
GSLHHDNHGQN
385
GSVFGVPSG
570
GSIAMTSHG
755

QQT

QQT

QNQQT

QNQQT

GHDSPHKSGQN
201
GIMARDSSGQN
386
GSGLPDRNL
571
GSPGVSPSG
756

QQT

QQT

QNQQT

QNQQT

GSGSPHARMQN
202
GVVHITNSGQN
387
GSGTHNSAIQ
572
GSGQNQQTG
757

QQT

QQT

NQQT

SSSRV

GSGSPHVKSQN
203
GSGQNQHSAPF
388
GSGMIIASMQ
573
GSGQHLPLL
758

QQT

NQT

NQQT

GNQQT

GQDSPHKSGQN
204
GSGQTSGLKQN
389
GGITWTDSG
574
GSDHSHRGG
759

QQT

QQT

QNQQT

QNQQT

GSGSPHASRQN
205
GSGQNQQTSLS
390
GSGQNQQAS
575
GSGIVTKLG
760

QQT

NTA

GRQQT

QNQQT

GSGSPHASRQN
206
GSGQNQAVHNK
391
GSGQNQQPH
576
GSGQDVTKT
761

KQT

SQT

LKSLT

GNQQT

GSGSPHVKIQNQ
207
GVHTHLPSGQN
392
GPPQHMTSG
577
GSGQNQQSH
762

QT

QQT

QNQQT

GRIGT

GSGSPHSKAKN
208
GHLTMHNSGQN
393
GSGQNQQAS
578
GSGQNQQIN
763

QQT

HQT

LPSRT

HRSPT

GSGSPHKKNQN
209
GSGSSSRPYQNQ
394
GSGQIVSTQT
579
GSGDDSRVG
764

QQT

QT

NQQT

QNQQT

GSGSPHVRMQN
210
GILLATPSGQNQ
395
GSGKGHSAG
580
GSGQSTLKRI
765

QQT

QT

QNQQT

NQQT

GSGSPHASRQK
211
GSGQNAGSFPN
396
GSGQNTRLQ
581
GSGSQHSKA
766

QQT

QQT

LGQQT

QNQQT

GHSSPHRSGQN
212
GSRDGHTVGQN
397
GSVGSRPVG
582
GSGQNQQHA
767

QQT

QQT

QNQQT

SSNNT

GMRTYHLSGQN
213
GSLLISTSGQNQ
398
GSSHTLALG
583
GSRTYQVSG
768

QQT

QT

QNQQT

QNQQT

GSGSPHTRGQN
214
GSGAMPSHGQN
399
GMYEYSQSG
584
GSGQNQGLL
769

QQT

QQT

QNQQT

SSPQT

GSGIIPVSSQNQ
215
GALVSPISGQNQ
400
GNGQNQQHS
585
GSGGGLQHN
770

QT

QT

ILHGT

QNQQT

GSEYGHKSGQN
216
GSLSSHGVGQN
401
GSGYNQPHL
586
GSGQNQQTT
771

QQT

QQT

QNQQT

AATRM

GRGQNVSSVHR
217
GSGQNQQASLA
402
GPLVNASSG
587
GSGQNQRAS
772

QQT

MRT

QNQQT

ILVQT

GSSHRFYGGQN
218
GPGLGSHSGQN
403
GSGQNQQVL
588
GSGQNLGLL
773

QQT

QQT

TTART

GAQQT

GYFVAAWSGQN
219
GHDSQHKSGQN
404
GSGQNQHSV
589
GSLDLGRSG
774

QQT

QQT

HNDQT

QNQQT

GSVLHSHAAQN
220
GSGLTLSATQN
405
GAGLIMHSG
590
GNSQVKVSG
775

QQT

QQT

QNQQT

QNQQT

GSGDLVVSTQN
221
GSGQVVAHVGN
406
GMGRHSASG
591
GSSGSHQYG
776

QQT

QQT

QNQQT

QNQQT

GSYGMAASGQN
222
GSGLRTMTTQN
407
GSHSQSGHG
592
GSGQNQQQR
777

QQT

QQT

QNQQT

DGTLT

GLNHFGASGQN
223
GSGQVGRLLQN
408
GSSTTIVSGQ
593
GRGQHVSVA
778

QQT

QQT

NHQT

NNQQT

GSTGSHSAGQN
224
GSGQLSHQSVN
409
GRHLVTASG
594
GDSSSRISGQ
779

QHT

QQT

QNQQT

NQQT

GLAGHTVSGQN
225
GSGDRYQTLQN
410
GSGQNQQHA
595
GSGQNQQHS
780

QQT

QQT

NLNQT

LSSQT

GIILGASSGQNQ
226
GSGQNQQLKSS
411
GSGSTHSKA
596
GSLMDVHRG
781

QT

AQT

QNQQT

QNQQT

GSGVSTYNIQNQ
227
GSGQNQYSIPVA
412
GSGQNKQML
597
GSIQYQSSGQ
782

QT

QT

SGNTT

NQQT

GSLVSVQTGQN
228
GSGERLHLTQN
413
GSGQVHNPT
598
GLGSKNPSG
783

QQT

QQT

QNQQT

QNQQT

GQSSPHRSGQN
229
GSGHNQQVRTA
414
GSGQNQQIP
599
GSGQLVLTL
784

QQT

PNT

HVHQT

QNQQT

GREYGHKSGQN
230
GGLSHVMSGQN
415
GSLHAGLSG
600
GSGQNQQTS
785

QQT

QQT

QNQQT

QPLPG

GHTLTLSSGQN
231
GSGQSHRDVLN
416
GPAQHGTSG
601
GSGQNQQNL
786

QQT

QQT

QNQQT

GKLNT

GSITLIPSGQNQ
232
GSGQNLAGRMD
417
GEKAVTSSG
602
GTTAHQPSG
787

QT

QQT

QNQQT

QNQQT

GSNGFTALGQN
233
GSGQNQQTNRG
418
GSGQNQQTM
603
GSGQNRAQI
788

QQT

NPM

ANGQR

GTQQT

GSGHSSHSVQN
234
GSGQSYQRDHN
419
GSGSPHSKD
604
GSGQYVHVS
789

QQT

QQT

QNQQT

SNQQT

GSGIPQRSGKNQ
235
GSLLSAGMGQN
420
GSFSMGYGG
605
GSGQNQQTA
790

QT

QHT

QNQQT

HAFNI

GSGDTLHMLQN
236
GSGQNQQTAIY
421
GSGTHLVSL
606
GSGQNQRTM
791

QQT

RNI

QNQQT

VATQT

GERHTVLSGQN
237
GSGQNQQTSGT
422
GSGQMQPHV
607
GSGQNPIRG
792

QQT

TNC

QNQQT

AMQQT

GSGMPQSHIQN
238
GMTSHSVSGQN
423
GSGQNQQVA
608
GSGYVITGS
793

QQT

QQT

GLNNT

QNQQT

GSGQLSGIGGNQ
239
GSSQSTGYQPN
424
GSSQNQQHD
609
GRGPKQSNI
794

QT

QQT

MRLRT

QNQQT

GSGQNRKPASF
240
GSLKPTTLGQN
425
GPASLPISGQ
610
GSGQNQQT
795

AQT

QQT

NQQT

MLGKPC

GSGSVSQLGQN
241
GRMFSLGSGQN
426
GSGQNQQPP
611
GSGQNQQVG
796

QQT

QQT

LATRT

STVRT

GSDFLGTHGQN
242
GSGQNQQTALG
427
GSSRVPVSG
612
GNVTTQKSG
797

QQT

VKC

QNQQT

QNQQT

GQIVQNPSGQN
243
GAMVSHSSGQN
428
GSGQNQQTN
613
GSGNPVSHL
798

QQT

QQT

LGHTT

QNQQT

GSGTQIPSQQNQ
244
GSGQNQQRNSD
429
GSGQNQQLV
614
GSLSHMESG
799

QT

SVT

SRVQT

QNQQT

GSGQNQQSARE
245
GSGQSMTLHLN
430
GPNSYPVSG
615
GRAPTNLSG
800

GLT

QQT

QKQQT

QNQQT

GSGLGMSTGQN
246
GSGQVHQAEVN
431
GHAHYQASG
616
GSGQNQQTV
801

QQT

QQT

QNQQT

MTARA

GSGLPVLSGQN
247
GSGQNQSQNHL
432
GSGQALLST
617
GSGMPASRL
802

QQT

QQT

GNQQT

QNQQT

GSGHSIRTDQNQ
248
GSLLTTASGQN
433
GSGQLPRQM
618
GVVRNHQSG
803

QT

QQT

TNQQT

QNQQT

GSGQSVQTVVN
249
GSGLIRTAAQNQ
434
GSGFPKSTEQ
619
GSGQNQHSV
804

QQT

QT

NQQT

QVRQT

GSGQNRAQSRF
250
GSGQNQQTVSR
435
GSRETSLSGQ
620
GSGQNTGHL
805

QQT

QST

NQQT

TMQQT

GGGDLGRSSQN
251
GSGQYANHGIN
436
GSGQNQQGT
621
GSGQNQQYA
806

QQT

QQT

GVSHT

GKILT

GGGTKMDSGQN
252
GSRSTGPSGQN
437
GSRTVPVYG
622
GSGNPHVRN
807

QQT

QQT

QNQQT

QNQQT

GSGSPHPSRQNQ
253
GRGVQQKLQQN
438
GSNAQSAHG
623
GSGQNGGSS
808

QT

QQT

QNQQT

NRQQT

GSGQFTNAGMN
254
GSGQNQQVHLS
439
GAFHLAASG
624
GSGQRLSQG
809

QQT

TGT

QNQQT

VNHQT

GGRNGHTVGQN
255
GSGQNQQLSAK
440
GSGQYRSSS
625
GSGQNAHAK
810

QQT

SST

DNQQT

EGQQT

GSGFGPQTGQN
256
GSGYKAARPQN
441
GSGQVYISTP
626
GSSPAPNSG
811

QQT

QQT

NQQT

QNQQT

GRTDSHTSGQN
257
GSAGISPSGQNQ
442
GSGVSTQLL
627
GLAHKTSSG
812

QQT

QT

QNQQT

QNQQT

GYEVLGSSGQN
258
GSGQNRAHAFL
443
GSGQLGLSV
628
GSGQNQQTP
813

QQT

QQT

TNQQT

GAHKT

GSVHLSVTGQN
259
GSGLSGITMQN
444
GSGSNMRLS
629
GSGQNQQSL
814

QQT

QQT

QNQQT

SGSFT

GFMSYKGSGQN
260
GPGSAHSSGQN
445
GSGQNLHSG
630
GSGQNQQST
815

QQT

QQT

LPQQT

GTSRT

GNIAGSVSGQN
261
GSSHTQALGQN
446
GSSHTLALG
631
GSGQNQQTV
816

QQT

QQT

QNKQT

QSNLV

GSGSHRDVSQN
262
GSGVHGVSSQN
447
GSGQNQHSL
632
GSGQNQQLG
817

QQT

QQT

PAHRT

SRQCT

GGLGSMSSGQN
263
GSSGRDMGGQN
448
GSGQNQGTV
633
GSGQNQYLR
818

QQT

QQT

YPNQT

LELQT

GSGHLPQSAQN
264
GERAFPTSGQN
449
GSGQNQQPS
634
GSGQNQQTS
819

QQT

QQT

LRQST

PRLQT

GGVLVGGSGQN
265
GGRIVSLSGQNQ
450
GSGQNARLK
635
GSGQNQQTT
820

QQT

QT

DNQQT

SSNMT

GTHPYTSSGQN
266
GSGQNSYSHTS
451
GHAGSTGSG
636
GTASTYNSG
821

QQT

QQT

QNQQT

QNQQT

GSGQNQQLKEN
267
GLGYPGSSGQN
452
GSGQALSSSG
637
GSGQNQQT
822

RST

QQT

NQQT

MPQHKI

GSGQNQQTSPH
268
GSGPQSHTGQN
453
GSGASESHR
638
GSGQSHLHT
823

NHT

QQT

QNQQT

GNQQT

GSGTLYPQSQN
269
GSGQNQQLSRD
454
GVGVITSSGQ
639
GVKGVGHSG
824

QQT

AST

NQQT

QNQQT

GSGQNQQSNWI
270
GSGQILHSVPNQ
455
GSLYGQSLG
640
GSGKVTKQS
825

TKT

QT

QNQQT

QNQQT

GSGYTSLFLQNQ
271
GSGFHTDSRQN
456
GSGQMSDVH
641
GSGQNQQTA
826

QT

QQT

GNQQT

LEKSL

GSGVMTHVLQN
272
GSGQSHSLATN
457
GSGQNQQHS
642
GSGYKDTYG
827

QQT

QQT

SKATT

QNQQT

GSVSDVRAGQN
273
GSGQNQQTLSK
458
GSGQNQQTS
643
GSGQNQQSG
828

QQT

PWT

VSQQT

TFLST

GSGQSHMATLN
274
GSGHAAISQQN
459
GSGQKMWK
644
GSGQNTGQH
829

QQT

QQT

LDNQQT

MMQQT

GSGLSVHLAQN
275
GSGQNQQQIGG
460
GSGQNVSMQ
645
GSGKNQQRP
830

QQT

NST

VNQQT

GLDQT

GSGLSHATQQN
276
GGGPMAGSGQN
461
GSGQNQRAT
646
GSGQSREISL
831

QQT

QQT

LSNQT

NQQT

GSGLSVQSGQN
277
GMRMEYQSGQ
462
GSGQASSKS
647
GTPTSPSSGQ
832

QQT

NQQT

ANQQT

NQQT

GSGHMTYREKN
278
GSGQNQQGTLL
463
GSGKNQTPIP
648
GKPAGGLSG
833

QQT

HQT

KGQT

QNQQT

GSKGVPTPGQN
279
GSGQNQRSSGG
464
GSGQNQQTR
649
GSGQNHRSA
834

QQT

VQT

QEGST

DMQQT

GSGLLPLSSQNQ
280
GSGQNQRGALA
465
GASSLATSG
650
GSGQNQQTL
835

QT

TQT

QNQQT

PSLSL

GNGLYAVSGQN
281
GSGTVHAATQN
466
GSGQRGSLT
651
GSPYMGATG
836

QQT

QQT

ENQQT

QNQQT

GFNGSPSSGQNQ
282
GSRMTQQFGQN
467
GSEQTRQRG
652
GSGHAKAVG
837

QT

QQT

QNQQT

QNQQT

GSGQIRHSDQN
283
GSSSPGASGQN
468
GSGQNQQTL
653
GHMKGVTS
838

QQT

QQT

TASKE

GQNQQT

GGQVAPSSGQN
284
GHPSPHVSGQN
469
GSGTSGKTG
654
GSGQNQKIL
839

QQT

QQT

KNQQT

TLDQT

GSFSMHTHGQN
285
GSGSHHASRQN
470
GQLVTFTSG
655
GSGQNQQTK
840

QQT

QQT

QNQQT

VGHSA

GSGQNQQVIQG
286
GAVGHSYSGQN
471
GSGQNQQSA
656
GIARTTISGQ
841

SNT

QQT

NKILT

NQQT

GRVLHSHAGQN
287
GSRSQYDIGQN
472
GSGQNQQHH
657
GSGQNQQTS
842

QQT

QQT

SSHTT

VGFRT

GSGQNQQTSLQ
288
GSGQGPQERGN
473
GSGQNQKG
658
GSGQNQQT
843

DQT

QQT

MQPNQT

MIANIR

GSGLGRAPVQN
289
GSIAHVGTGQN
474
GSGQLVSGL
659
GDMTRSSSG
844

QQT

QQT

YNQQT

QNQQT

GNGFSSASGQN
290
GSGQNQQKQNH
475
GSSVGVPSG
660
GSGHMSDLR
845

QQT

GNT

QNQQT

QNQQT

GSGQMASRESN
291
GSGQNQQALGS
476
GSGQNQQW
661
GRGAVMAS
846

QQT

QRT

DSRRQT

GQNQQT

GPGLPNHSGQN
292
GSGAITHMPQN
477
GSEQTRQSG
662
GSGQNQQLS
847

QQT

QQT

QNQQT

GKSVT

GNIQWQGSGQN
293
GSGQRNPLLLN
478
GSGIGSHIPQ
663
GSHTLVVSG
848

QQT

QQT

NQQT

QNQQT

GMSAHMSSGQN
294
GSSGIPVSHQNQ
479
GSGQNQRLH
664
GSGPWSAGL
849

QQT

QT

GVDQT

QNQQT

GHSFVNRSGQN
295
GVHSLTPSGQN
480
GEVSRVLSG
665
GSGQHSPHA
850

QQT

QQT

QNQQT

LNQQT

GRAVMDHSGQ
296
GVIVLHGSGQN
481
GSGQNQQKV
666
GSGQNQQPN
851

NQQT

QQT

SPLLT

SGSMT

GALTVMQSGQN
297
GGTRVVDSGQN
482
GSGLALERS
667
GSGLAHLGG
852

QQT

QQT

QNQQT

QNQQT

GSGQRSPVLPNQ
298
GSGGVTYQSQN
483
GPDRIGSSGQ
668
GSSVRYEPK
853

QT

QQT

NQQT

QNQQT

GSGQNGHLSLK
299
GSGQNQAGHGP
484
GSGQNQDHQ
669
GSGQNQQAR
854

QQT

GQT

NKQQT

PLELT

GSLPRGTSDQN
300
GSGQLVTSGPN
485
GSGQNQQTA
670
GSGQPRSTGI
855

QQT

QQT

LYNNT

NQQT

GVAGSLVSGQN
301
GSGIAAQRTQN
486
GSGAVHLTA
671
GSGQNQAN
856

QQT

QQT

QNQQT

WVKVQT

GRGGIPQSGQN
302
GSTPAGVGGQN
487
GSLVSTQSG
672
GSGHLFQSG
857

QQT

QQT

QNQQT

QNQQT

GSGQYASSIPNQ
303
GSGQNQQTSTG
488
GSGVSARMV
673
GSGQNRGISI
858

QT

VHS

QNQQT

SQQT

GTDFGRQSSQN
304
GSGQIRQLVDN
489
GSGQTRMPL
674
GSGTHYDNR
859

QQT

QQT

ANQQT

QNQQT

GIFMQTPSGQNQ
305
GSLIGMQSGQN
490
GSGISSRNMQ
675
GSGQNQQTS
860

QT

QQT

NQQT

TTPLP

GSGQNQQTRLV
306
GSGQIKGKMDN
491
GSGEKVHSG
676
GSGQVHASQ
861

DLT

QQT

QNQQT

VNQKT

GTREMPLSGQN
307
GSGSDMSSWQN
492
GSGQNQQKL
677
GSSGHRESG
862

QQT

QQT

SSMST

QNQQT

GSRLVHVHGQN
308
GRGQNQQHTGL
493
GSGQNQQTG
678
GLSAEKSSG
863

QQT

ATT

QHMRV

QNQQT

GSGRLVPNGPN
309
GSGQNQQTLYS
494
GSGMIHTTA
679
GSGQEHRSL
864

QQT

SNT

QNQQT

ANQQT

GSGYLRESPQN
310
GSGQTQVLKSN
495
GSGQNWPAL
680
GSGQTVVRI
865

QQT

QQT

KGQQT

ANQQT

GARIQNASGQK
311
GSRTLSNVGQN
496
GASHMSISG
681
GSGQNVSSV
866

QQT

QQT

QNQQT

HRQQT

GLSNPMPSGQN
312
GSGVQHSLPQN
497
GSDQNQQLG
682
GSGASRMSI
867

QQT

QQT

YSKQT

QNQQT

GSTVQDTRGQN
313
GNYLHQASGQN
498
GIPSIRESGQ
683
GVAFIGSSGQ
868

QQT

QQT

NQQT

NQQT

GPFGMPSSGQN
314
GSGGTSVHQQN
499
GSGIPSVKFQ
684
GSGQNQQTV
869

QQT

QQT

NQQT

PTRQT

GSGQNHGVLSN
315
GMDHSRPSGQN
500
GSGQNQQTS
685
GSGQAAKSS
870

QQT

QQT

VSQNV

QNQQT

GSGYSMSQAQN
316
GSGQNQQSMGT
501
GSGQNQQIG
686
GSGQNQQVA
871

QQT

FTT

ESRMT

IRTST

GSGMLTHTLQN
317
GSGQNQQTPLR
502
GSGSSSMSFQ
687
GSVHMQNA
872

QQT

PPT

NQQT

GQNQQT

GRGSPHASRQN
318
GSGQNQHHSVS
503
GSGQKQERA
688
GSGMRQAG
873

QQT

QQT

VSKQT

VQNQQT

GLSWPSTSGQN
319
GSGQLRSLSTNQ
504
GCTTRLNSG
689
GSGQNQQVG
874

QQT

QT

QNQQT

GKTVT

GNSMERTSGQN
320
GSGSPRQLSQN
505
GSGQNQQIIS
690
GVHDMRVS
875

QQT

QQT

TKIT

GQNQQT

GSGMSPSTLQN
321
GSGQNQQTTAS
506
GSGQNQQKS
691
GSGQHVSVA
876

QQT

SHT

LNGNT

NNQQT

GSGHGQVLSQN
322
GRGQVVSTHQN
507
GSGIPAPRLQ
692
GSAAMSVRG
877

QQT

QQT

NQQT

QNQQT

GRGQIYSTGGN
323
GSAQVSMVGQN
508
GSGQIRESM
693
GVSRGGPSG
878

QQT

QQT

GNQQT

QNQQT

GVVAAHNSGQN
324
GSSTLVTIGKNQ
509
GSGQNSGVH
694
GSGQMVHTI
879

QQT

QT

FNQQT

GNQQT

GDSSLRHSGQN
325
GFAHQASSGQN
510
GSGQNIGHSL
695
GRGGSMAET
880

QQT

QQT

PQQT

QNQQT

GSLVSQGAGQN
326
GSGQPVLSISNQ
511
GSGERSISVQ
696
GSGHTNPTR
881

QQT

QT

NQQT

QNQQT

GSLLQAHSGQN
327
GSGQSHRSELN
512
GSGLKPNVL
697
GSGEAARYE
882

QQT

QQT

QNQQT

QNQQT

GSGHIYVGIQNQ
328
GSSVGSPIGQNQ
513
GSGQVAYAQ
698
GSGQNERHL
883

QT

QT

GNQQT

VLQQT

GHHTTVQSGQN
329
GSGMPIRNVQN
514
GSGQSSYGS
699
GSGQNQQSK
884

QQT

QQT

GNQQT

QQVLT

GSRQSKRNELN
330
GSSTRVDSGQN
515
GSGQNQAMT
700
GSGQARAHR
885

QQT

QQT

HGDQT

GNQQT

GSGQNQQHVSS
331
GSGQNQQTAMR
516
GSGQNQALV
701
GSGQNQQPL
886

PRT

STT

SMGQT

DTSRT

GSSKELLWGQN
332
GSGQNQQHSSS
517
GSGQNPSFM
702
GSGQNQQLA
887

QQT

HLT

RGQQT

NMVTT

GSLSTPSSGQNQ
333
GSRNGHAVGQN
518
GSGQNQQSH
703
GSGQMKDL
888

QT

QQT

LRTNT

HRNQQT

GSIGYAGQGQN
334
GLGAYQSSGQN
519
GYTRLETSG
704
GSGQNQHLS
889

QQT

QQT

QNQQT

SFVQT

GSGQNQRVSNS
335
GPGLSGHSGQN
520
GSGQSYDMR
705
GSGQNQQPS
890

QQT

QQT

GNQQT

SRVTT

GSGYASHVQQN
336
GSTGIVSSGQNQ
521
GSRTTQDIGQ
706
GSGQNQQLA
891

QQT

QT

NQQT

ITLGT

GSGEYSRSGQN
337
GSRTTQVIGQNQ
522
GSGHPYKAA
707
GSGQNQQTV
892

QQT

QT

QNQQT

GNPAT

GSVSTHSSGQN
338
GSGLLHRAQQN
523
GRLSNAHGG
708
GSGQNQGRA
893

QQT

QQT

QNQQT

HPMQT

GSGQNQHSLGN
339
GSGQNAQQAAA
524
GSGQNQRAV
709
GSGQLIASV
894

YQT

QQT

LNDQT

VNQQT

GSGGLDTRGQN
340
GSGQNQQSALR
525
GGSHTYGGG
710
GSSVRSLVG
895

QQT

TQT

QNQQT

QNQQT

GNILHATSGQN
341
GSGFLSDTRQN
526
GSSVNSMIG
711
GGAGSAHSG
896

QQT

QQT

QNQQT

QNQQT

GSGQSYTMTQN
342
GSGLLYHDQQN
527
GNSSMMGSG
712
GSDQNQQT
897

QQT

QQT

QNQQT

MSSTRT

GSGQNQHSAPN
343
GSGQNQHYSLH
528
GNRDRPSSG
713
GSGQNQQM
898

SQT

KQT

QNQQT

AGAFRT

GSGQNQQTMDH
344
GSGHSPLPQQN
529
GSGNMHASR
714
GSLGNLQRG
899

NRT

QQT

QNQQT

QNQQT

GSNGGVGTGQN
345
GNGHSMRPNQN
530
GFIFPKVSGQ
715
GSGPSISHGQ
900

QQT

QQT

NQQT

NQQT

GAGSIIPSGQNQ
346
GSGLKWSTLQN
531
GSGQNQQLK
716
GSGQNQQSS
901

QT

QQT

NSTST

FNVQT

GSGQTHGGQHN
347
GSGQMGRQAV
532
GSGQNQQSQ
717
GSGQNQQTG
902

QQT

NQQT

YMPRT

QATHN

GSNLSFQSGQN
348
GSGQNQQTSGV
533
GSGQRMADI
718
VSGSPHSKA
903

QQT

LTL

GNQQT

QNQQT

GATLQVHSGQN
349
GSGQNQQALHN
534
GSGQNQSHY
719
CSGSPHSKA
904

QQT

PHT

PSQQT

QNQQT

GSGFNQRSEQN
350
GSGQNQQVIPNS
535
GSDGKMHRG
720
GSGSPHRKA
905

QQT

KT

QNQQT

QNQQT

GSGSLRDFDQN
351
GSPLQDRVGQN
536
GSGSVGFIGQ
721
GRGSPHSKA
906

QQT

QQT

NQQT

QNQQT

GSGDSITGKQNQ
352
GSGQNQYSSTN
537
GLHGMTLSG
722
GSGSPHSKA
907

QT

PQT

QNQQT

QNKQT

GSGQDRNIVQN
353
GAMTVTISGQN
538
GSDQSKRGD
723
GSGSPHSKA
908

QQT

QQT

SNQQT

QTQQT

GSGLSHSHQQN
354
GSGQNQQLQTLI
539
GSLFLATGG
724
GSGSTHASR
909

QQT

RT

QNQQT

QNQQT

GSGQNQQTGMS
355
GSGLRQTSQQN
540
GSGQNQQPS
725
GSGSPHKYG
910

SVK

QQT

AFSKT

QNQQT

GSVTHGISGQNQ
356
GSGQNQQTGLR
541
GSGQLPQSG
726
GSGSPHKFG
911

QT

QQT

LNQQT

QNQQT

GVVAHQPSGQN
357
GSGQTRQMKDN
542
GSGSKQNAL
727
VSGSPHKFG
912

QQT

QQT

QNQQT

QNQQT

GSGPILGQLQNQ
358
GSGQNHGLQSG
543
GSGQRRELS
728
GSGSPHSKA
913

QT

QQT

QNQQT

QNHQT

GSGHVPNSGLN
359
GSGQSHRQPEN
544
GSGQREPKA
729
GSGSPHSKA
914

QQT

QQT

SNQQT

QHQQT

GDAGVRSSGQN
360
GSGQDRHIVQN
545
GSGQNQQHP
730
GSGSPHKTY
915

QQT

QQT

STQQT

QNQQT

GSGSQLMSLQN
361
GSGQNQQLPHS
546
GSQSTLGLG
731
VSGSPHASR
916

QQT

NLT

QNQQT

QNQQT

GSGLDYSQRQN
362
GSGQLSVPYDN
547
GSGQNQQMP
732
GSGSPHKFG
917

QQT

QQT

GLSST

KNQQT

GSGQSSGRLINK
363
GSGRNPQTQPL
548
GSGQNQQTV
733
GSGSPHASR
918

QT

QQT

GGKNL

QNQHT

GSSVSPSSGQNQ
364
GSGQPYSTGLN
549
GSSREFHSGQ
734
GSHSPHKSG
919

QT

QQT

NQQT

QNQQT

GSGQVVGLSGN
365
GSGQNQQTHGG
550
GSGQNQQTV
735
GSGQNQQRR
920

QQT

LRD

PSNLV

MSPST

GSNMGVPLGQN
366
GAYGMVSSGQN
551
GSGQNAYSS
736
GSGSPHSKP
921

QQT

QQT

QAQQT

QNQQT

GSFYPSSTGQNQ
367
GSGIQSSYSQNQ
552
GSGQNKDHS
737
GSGSPHKFG
922

QT

QT

TRRQT

QKQQT

GSGQNQQTRLT
368
GPRLSDQSGQN
553
GQLGSVGSG
738
VSGSPHGAR
923

DLT

QQT

QDQQT

QNQQT

GPTNGRSSGQN
369
GSGQNQQTHPS
554
GSGQHAAPG
739
GSGSPHSKA
924

QQT

PCT

HNQQT

QKQQT

GSGLLHGKLQN
370
GSGQSFQMHTN
555
GSGQNQQTS
740
GSHSPHKRG
925

QQT

QQT

QSPPT

QNQQT

GANMGHVSGQ
371
GSGQNQQTGNP
556
GSGNYRDHE
741
GSGQNRQRL
926

NQQT

KHT

QNQQT

KGLET

GSGQNQQSGRG
372
GFSSAVHSGQN
557
GSGQHSNQH
742
GSGSPHKLG
927

DLT

QQT

VNQQT

QNQQT

GSHGHYASGQK
373
GSGQNQQTSMS
558
GSGQTARNG
743
GSGSPHKTS
928

QQT

NAT

INQQT

KNQQT

GSGDLRISPQNQ
374
GSGQDMKQHH
559
GSGQNQQHY
744
GSGSPHKIGQ
929

QT

NQQT

GSQGT

NQQT

GSGMPVILGQN
375
GLRLSTPSGQNQ
560
GSGSPQASR
745
GSGQDSPHV
930

QQT

QT

QNQQT

RNQQT

GRGVITSSGQNH
376
GSGQNQQTSVY
561
GSGFSHSMG
746
GSGSPHKTS
931

QT

MNT

KNQQT

QNQQT

GSGHSVSGPQN
377
GSGQNQYSQSS
562
GSGQSHSLET
747
GSGSPHASR
932

QQT

MQT

NQQT

KNQQT

GSRNGHTVGRN
378
GSGQNQQSMAD
563
GTEQTRQSG
748
GSHSPHKSG
933

QQT

HTT

QNQQT

QKQQT

GAGVHMVSGQ
379
GWERSFVSGQN
564
GSGRHLASV
749
GSGSPHKTS
934

NQQT

QQT

QNQQT

QKQQT

GSGQNHRPSVL
380
GLLAGKSSGQN
565
GLGSKNHSG
750
GSGSPHVRG
935

QQT

QQT

QNQQT

QNKQT

GSGSPRDSIQNQ
381
GKSFVPQSGQN
566
GSGQNQQTS
751
GSGSPHKTT
936

QT

QQT

HFPSA

QNQQT

GSGQGIHSSVNQ
382
GSGQMQSAGSN
567
GSGQLSGTP
752
GSGPVRALR
937

QT

QQT

QNQQT

QNQQT

GSGQQLSITPNQ
383
GSDQNQRLTSS
568
GSGQNQQAP
753
GSGSPHVRG
938

QT

MQT

HKKET

QKQQT

GGYHSQTSGQN
384
GESRAVLSGQN
569
GSGQNQQTL
754
CSGSPHKTS
939

QQT

QQT

RGSLE

QNQQT

CSHSPHKSGQN
940
DAGSPHSKAQN
1909
GSGSPHASR
2019
GNDSPHKSV
2129

QQT

QQ

QNQQ

QNQQ

GHDSPHKSGQN
1800
GSGSPHSKGQN
1910
GSGSPHASR
2020
GHDSPHKSA
2130

QQ

QQ

QNKQ

QNYQ

GSGSPHSKAQN
1801
DGGSPHSKAQN
1911
GSGSPHVKIQ
2021
GSASPHSKA
2131

QQ

QQ

NQQ

LNQQ

GSGSPHSKAQN
1802
ASGSPHSKAHN
1912
GSGSPHSKA
2022
GHESPHKSA
2132

RH

KNQQ

QNRQ

GSGSPHSKVQN
1803
GSGSPHSKAQN
1913
GSGSPHKKN
2023
GQDSPHKIG
2133

QQ

TY

QNQQ

QNQQ

MSGSPHSKAQN
1804
GSGSPHSKSQN
1914
GSGSPHVRM
2024
GHDSPHKSG
2134

QQ

QH

QNQQ

QNHL

GRGSPHSKAQN
1805
GGGSPHSKAQD
1915
GSGSPHASR
2025
GHDSPHKSG
2135

QQ

KQ

QKQQ

QYQH

RNGSPHSKAQN
1806
GSGSPHSKAQN
1916
GHSSPHRSG
2026
GNDSPHKSV
2136

QQ

HL

QNQQ

QNHQ

GSGSPHSKARD
1807
GSGSPHSKAQIG
1917
GSGSPHTRG
2027
GHDSPHKSG
2137

QQ

M

QNQQ

QNQW

GSGSPHSKAPNL
1808
GSGSPHSKALN
1918
CSGSPHSKA
2028
GHDSPHKSV
2138

Q

KQ

QNQQ

QNQH

TSGSPHSKAQN
1809
GGGSPHSKAQN
1919
GSGSPHRKA
2029
GHDSPHKSG
2139

QQ

PQ

QNQQ

QNQH

GSGSPHSKAHV
1810
GTGSPHSKAPN
1920
GSGSPHSKA
2030
GHDSPHKSG
2140

RQ

QL

QNKQ

QTRQ

GSGSPHSKAPNQ
1811
GSGSPHSKAQL
1921
GSGSPHSKA
2031
GHDSPHKSG
2141

H

QQ

QTQQ

QNLH

ISGSPHSKAQNQ
1812
GGGSPHSKAQY
1922
CSGSPHKTSQ
2032
GHDSPHKSA
2142

Q

QQ

NQQ

QNQE

GPGSPHSKAHN
1813
GGGSPHSKAQH
1923
CSHSPHKSG
2033
GHDSPHKSG
2143

QQ

QQ

QNQQ

QHLQ

GSGSPHSKTQSQ
1814
GSGSPHSKAQR
1924
GQSSPHRSG
2034
GHDSPHKSR
2144

Q

MS

QNQQ

LNQP

ESGSPHSKAQN
1815
GSGSPHSKAQGI
1925
GRGSPHASR
2035
GQDSPHKSG
2145

QQ

L

QNQQ

QNQD

GSGSPHSKAQPA
1816
GSGSPHSKAQD
1926
GSGSPHASR
2036
GHDSPHKSG
2146

K

RQ

KNQQ

RNQQ

SSGSPHSKAQNQ
1817
GSGSPHSKARD
1927
GSGSPHASR
2037
GHDSPHKSG
2147

Q

WQ

QNQH

QNLL

GNGSPHSKAQN
1818
GSGSPHSKAQN
1928
GSGSPHKFG
2038
GHDSPHKSG
2148

QQ

TH

KNQQ

QLVI

GSGSPHSKSQTQ
1819
GSGSPHSKAQE
1929
GSGSPHKFG
2039
GHDSPHKSR
2149

Q

RS

QKQQ

QSQQ

ASGSPHSKAQN
1820
GSGSPHSKAQN
1930
GSGSPHKFG
2040
GHDSPHKSG
2150

QQ

YQ

QNQQ

RTQE

GSGSPHSKAQN
1821
GSGSPHSKAQR
1931
GSGSPHKIGQ
2041
GHDSPHKSV
2151

LA

TC

NQQ

QTHQ

GSGSPHSKSQNQ
1822
GSGSPHSKAQIG
1932
GSGSPHKLG
2042
GHDSPHKSG
2152

L

H

QNQQ

QNQP

NSGSPHSKAQN
1823
GSGSPHSKAQG
1933
GSGSPHKTSK
2043
GHDSPHKSG
2153

QQ

AI

NQQ

QTQQ

GSGSPHSKAQG
1824
GSGSPHSKAQV
1934
GSGSPHKTSQ
2044
GPDSPHKIGQ
2154

QQ

PP

KQQ

NQQ

VSGSPHSKAQN
1825
GSGSPHSKAQV
1935
GSGSPHKTSQ
2045
GHDSPHKSV
2155

QQ

QQ

NQQ

QNQQ

GSGSPHSKALNR
1826
GSGSPHSKALM
1936
GSGSPHKTT
2046
GHDSPHKSR
2156

Q

RQ

QNQQ

QDQH

LSGSPHSKAQN
1827
GSGSPHSKAQY
1937
GSGSPHKTY
2047
GPDSPHKSG
2157

QQ

SV

QNQQ

QKQQ

GSGSPHSKAHN
1828
GSGSPHSKVPNL
1938
GSGSPHKYG
2048
GHDSPHKSR
2158

QQ

0

QNQQ

QSQH

GSGSPHSKTQN
1829
GSGSPHSKAQAI
1939
GSGSPHSKA
2049
GHDSPHKSV
2159

QQ

T

QHQQ

QNQL

GGGSPHSKAQT
1830
GSGSPHSKAQK
1940
GSGSPHSKD
2050
GYDSPHKSG
2160

QQ

TL

QNQQ

QYQH

GSGSPHSKAQNP
1831
GSGSPHSKAQN
1941
GSGSPHSKPQ
2051
GHDSPHKSR
2161

P

QW

NQQ

QNQQ

GSGSPHSKAQN
1832
GSGSPHSKAQL
1942
GSGSPHVRG
2052
GHDSPHKSW
2162

LQ

HH

QKQQ

VRQQ

GGGSPHSKAQN
1833
GSGSPHSKAQNI
1943
GSGSPHVRG
2053
GHESPHKSG
2163

QQ

I

QNKQ

QNQH

GSGSPHSKAQY
1834
MEGSPHSKAQN
1944
GSHSPHKRG
2054
GHDSPHKIG
2164

QQ

QQ

QNQQ

HNQQ

GGGSPHSKAQN
1835
GSGSPHSKAQG
1945
GSHSPHKSG
2055
GHDSPHKSN
2165

KQ

HH

QKQQ

AWQQ

GSGSPHSKAQD
1836
GSGSPHSKAQS
1946
GSHSPHKSG
2056
GHDSPHKSG
2166

QE

KV

QNQQ

QSVP

KSGSPHSKAQN
1837
GSGSPHSKAQLP
1947
VSGSPHASR
2057
GHESPHKSG
2167

QQ

S

QNQQ

QNIQ

GGGSPHSKAQN
1838
GSGSPHSKAIGK
1948
VSGSPHGAR
2058
GHDSPHKSV
2168

QL

Q

QNQQ

QNHL

GSGSPHSKAQN
1839
GGGSPHSKSQN
1949
VSGSPHKFG
2059
GHDSPHKIG
2169

HQ

QQ

QNQQ

LDQQ

GSGSPHSKAQD
1840
GSGSPHSKAQAI
1950
GSGSPHSKA
2060
ASGSPHSKA
2170

QQ

H

QYYV

QHQQ

GGGSPHSKSQN
1841
GSGSPHSKAQH
1951
GSGSPHSKLR
2061
GHDSPHKRG
2171

QL

GL

RQQ

PDQQ

GNGSPHSKAQN
1842
GSGSPHSKAQF
1952
GSGSPHSKA
2062
GMGSPHSKT
2172

KQ

MC

GCGQ

QNQQ

GSGSPHSKGHW
1843
VSGSPHSKAQG
1953
GSGSPHSRA
2063
GHDSPHKSG
2173

QQ

QQ

QNQQ

ESQQ

GSGSPHSKAPNQ
1844
GGGSPHSKAQN
1954
GSGSPHSKRL
2064
GHDSPHKHG
2174

Q

QM

RQQ

QNHQ

GSGSPHSKAQN
1845
GSGSPHSKAQH
1955
GSGSPHSLRR
2065
GTGSPHSKA
2175

QL

LQ

NQQ

QNQL

GSGSPHSKRPEQ
1846
ENGSPHSKAQN
1956
GSGSPHSRGR
2066
GHDSPHKSV
2176

Q

QQ

NQQ

QNKQ

GSGSPHSKAQRT
1847
GSGSPHSKTQN
1957
GSGSPHSSRR
2067
GQVSPHKSG
2177

M

HQ

NQQ

QNQQ

GNGSPHSKAQN
1848
GSGSPHSKAQP
1958
GSGSPHSKAF
2068
GHDSPHKSG
2178

QH

AR

RLQ

QRQL

HSGSPHSKAQN
1849
GSGSPHSKAQSL
1959
GSCSPHRKA
2069
GHDSPHKIG
2179

QQ

Q

QNQQ

QNQL

GGGSPHSKALN
1850
GSGSPHSKSQSQ
1960
GSGSPHFLRQ
2070
GHDSPHKSG
2180

QQ

L

NQQ

QIIV

GSGSPHSKALH
1851
GSASPHSKAHS
1961
GSGSPHSLRF
2071
GYDSPHKSG
2181

QH

QQ

NQQ

QKQS

GTGSPHSKAQN
1852
GSGSPHSKAQM
1962
GSGSPHSKW
2072
GNGSPHSKA
2182

HQ

PS

LLQQ

QNQE

GSGSPHSKAQH
1853
GSGSPHSKAQG
1963
GSGSPHSKRR
2073
GDDSPHKSV
2183

RI

SL

LQQ

QNQQ

GSGSPHSKAQYI
1854
GSGSPHSKSQN
1964
GSGSPHSKA
2074
GHDSPHKSV
2184

H

QQ

QRKL

QSHQ

GGGSPHSKAHN
1855
GNGSPHSKSQN
1965
GSGSPHSKAL
2075
GHDSPHKSG
2185

QQ

QQ

RRQ

QFVV

GSGSPHSKAQKF
1856
GSGSPHSKAQV
1966
GSGSPHSKA
2076
GHDSPHKSR
2186

E

PA

QRLR

QNLQ

ESGSPHSKAQN
1857
GNGSPHSKAQN
1967
GSGSPHSKA
2077
GHNSPHKSG
2187

HQ

LQ

QRRL

QNQE

GSGSPHSKAQFP
1858
GSGSPHSKAQD
1968
GSGSPHSKA
2078
GHDSPHKSG
2188

S

KQ

RRQQ

QSQP

PSGSPHSKAQNQ
1859
GSGSPHSKAHY
1969
GSGSPHSKA
2079
GHESPHKSG
2189

Q

QQ

RRLQ

QNEQ

GNGSPHSKAQN
1860
GSGSPHSKAQV
1970
GSGSPHSKSR
2080
GHDSPHKSG
2190

PL

PS

RQQ

QNQL

GGGSPHSKAQS
1861
GGGSPHSKAQN
1971
GLLSPHWKA
2081
GHDSPHKSA
2191

QQ

HQ

QNQQ

QNLL

GSGSPHSKAQAI
1862
GSGSPHSKARG
1972
GSGSPHSKA
2082
ASGSPHSKAI
2192

K

EQ

RLRQ

NQQ

GSGSPHSKGQN
1863
GGGSPHSKAQY
1973
GSGSPHSKAS
2083
GNGSPHKRG
2193

RQ

QH

KRQ

QNQQ

GSGSPHSKAQSQ
1864
GSGSPHSKAPG
1974
GSGSPHVRR
2084
GHDSPHKSG
2194

Q

QQ

QNQQ

QSLQ

GSVSPHGKAQN
1865
KNGSPHSKAQN
1975
GSGSPHSKA
2085
GHDSPHKSA
2195

QL

QQ

QLYR

QNHQ

ASGSPHSKAQN
1866
GSGSPHSKRLEQ
1976
GSGSPHSKA
2086
GHDSPHKSG
2196

QL

Q

QLTV

RNRQ

RSGSPHSKAQN
1867
GSGSPHSKAQN
1977
GHDSPHKRG
2087
GHDSPHKYG
2197

QQ

QS

QHRQ

QNEQ

GSGSPHSKAQY
1868
GSGSPHSKAQK
1978
GHDSPHKSG
2088
GNGSPHSKA
2198

QH

VI

QKQQ

PNLQ

GSGSPHTKAQN
1869
GSGSPHSKAQN
1979
GHDSPHKSG
2089
GHDSPHKSQ
2199

PQ

ND

LTQQ

QNQQ

GSGSPHSKGQNP
1870
GSGSPHSKAQS
1980
GDDSPHKSG
2090
GHDSPHKSV
2200

P

VH

RNQQ

QSKQ

GSGSPHSKAQH
1871
GSGSPHSKAQPL
1981
GHDSPHKSG
2091
GNDSPHKIG
2201

QL

G

LNQQ

HNQQ

GSGSPHSKAQSP
1872
KEGSPHSKAQN
1982
GHESPHKSA
2092
GGGSPHSKA
2202

P

QQ

QNHQ

QDQQ

GSGSPHSKAQA
1873
GSGSPHSKAHN
1983
GHDSPHKSA
2093
GQDSPHKSG
2203

KL

QE

QNQW

QNPL

GSGSPHSKTKSQ
1874
GSGSPHSKAQIQ
1984
GHDSPHKSG
2094
ASGSPHSKA
2204

Q

Q

QNTH

QNHQ

GSGSPHSKAQD
1875
GSGSPHSKAQV
1985
GHDSPHKSG
2095
GHDSPHKSG
2205

RP

RN

RRRQ

RDQK

GIGSPHSKAQNL
1876
GSGSPHSKAPSN
1986
GHDSPHKSA
2096
GHDSPHKSV
2206

G

Q

QNQQ

HNQQ

GSGSPHSKAQAF
1877
GSGSPHSKAQV
1987
GHDSPHKSG
2097
GHDSPHKSG
2207

H

GH

QYQQ

QWKR

GSGSPHSKAQK
1878
GSGSPHSKAQR
1988
GNYSPHKIG
2098
GSGSPHSKA
2208

QQ

DI

QNQQ

ENRQ

GSGSPHSKAQN
1879
GSGSPHSKAQM
1989
GHDSPHKSR
2099
GHDSPHKSG
2209

AQ

PN

QNDQ

QSQQ

WSGSPHSKAQN
1880
AIGSPHSKAQNQ
1990
GHDSPHKSG
2100
GHDSPHKSR
2210

QQ

Q

QIRQ

QAQQ

GSGSPHSKAHN
1881
GSGSPHSKARG
1991
GHDSPHKIG
2101
GHDSPHKSV
2211

QL

LQ

QNQH

QNHQ

GNGSPHSKAQN
1882
GSGSPHSKLQK
1992
GYDSPHKSG
2102
GHDSPHKSK
2212

HQ

QQ

QKQQ

QNQQ

GGGSPHSKAQN
1883
GSGSPHSKAPSL
1993
GHDSPHKSG
2103
GHDSPHKSA
2213

LQ

Q

QSVQ

QNQL

GSGSPHSKAQK
1884
GSGSPHSKAQR
1994
GHESPHKSG
2104
GHDSPHKSG
2214

LN

DQ

RSQQ

QTQP

GGGSPHSKSQN
1885
GSGSPHSKNRD
1995
GHDSPHKSG
2105
GHDSPHKLW
2215

QH

QQ

QNKL

INQQ

GSGSPHSKSQNV
1886
GSGSPHSKAQA
1996
GHDSPHKTG
2106
GPDSPHKSG
2216

Q

KG

QNQQ

QNQQ

GSGSPHSKAQA
1887
GSGSPHSKAQS
1997
GRGSPHKRG
2107
GHDSPHKSV
2217

QQ

AH

QNQQ

QKQL

DSGSPHSKAQN
1888
GNGSPHSKSQN
1998
GSGSPHTKA
2108
GHPSPHWKG
2218

QQ

QH

QNPP

QNQQ

ASGSPHSKAPNQ
1889
GSGSPHSKSQN
1999
GQDSPHKSG
2109
GHDSPHKSG
2219

Q

HQ

QHQQ

RNQL

GSGSPHSKAQTP
1890
RSGSPHSKAQD
2000
GHDSPHKSG
2110
GSGSPHSKV
2220

P

QQ

QIQH

QDQQ

IDGSPHSKAQNQ
1891
GSGSPHSKAQST
2001
GHDSPHKSG
2111
GHDSPHKMG
2221

Q

M

PRQQ

RNQQ

GSGSPHNKAQN
1892
GSGSPHSKAQR
2002
GHDSPHKSG
2112
GHDSPHKSGI
2222

HQ

EM

HTQQ

SIQ

GSGSPHSKAQPP
1893
GGGSPHSKSQN
2003
GHDSPHKSG
2113
GHDSPHKSV
2223

A

RQ

QRQH

QNLQ

GSGSPHSKAQER
1894
GSGSPHSKAQY
2004
GSGSPHTKA
2114
GHDSPHKMA
2224

P

RA

QNQQ

HNQQ

GSGSPHSKAQD
1895
GGGSPHSKAQR
2005
GHDSPHKSA
2115
GHDSPHKHG
2225

LQ

QQ

QSQQ

QNQQ

GGGSPHSKAQN
1896
GSGSPHSKNQW
2006
GHESPHKSG
2116
GHDSPHKSV
2226

PP

QQ

QNQQ

QSQQ

GSGSPHSKAQA
1897
GSGSPHSKAQR
2007
GHDSPHKSL
2117
GHDSPHKSG
2227

MH

MN

QNQQ

QTVC

GSGSPHSKALN
1898
GSGSPHAKAQN
2008
GHGSPHSKA
2118
GQDSPHKSG
2228

QQ

HQ

QNPQ

QYQQ

GSGSPHSKAQHP
1899
GSGSPHSKAGD
2009
GHDSPHKSG
2119
GHDSPHKSG
2229

S

SQ

RNQE

QQIM

GLGSPHSKSQN
1900
GSGSPHSKLKSQ
2010
GHDSPHKSG
2120
GHDSPHKSR
2230

QQ

Q

QTQL

QNEQ

GTGSPHSKAQN
1901
GSGSPHSKAQKI
2011
GHDSPHKSE
2121
GHDSPHKSG
2231

QQ

S

KNQQ

LNHQ

GSGSPHSKAPGL
1902
GSGSPHSKAPS
2012
GRDSPHKSG
2122
GYDSPHKSG
2232

Q

MQ

QDQQ

QNQQ

GSGSPHSKAQGI
1903
GSGSPHSKASPR
2013
GHDSPHKTG
2123
GHDSPHKSG
2233

R

Q

HNQQ

QNLQ

GSGSPHSKAQAP
1904
GSGSPHSKRME
2014
GYDSPHKSG
2124
GHDSPHKSR
2234

A

QQ

QTQQ

QDQQ

GSGSPHSKSQSQ
1905
GSGSPHSKAQY
2015
GHESPHKSG
2125
GDDSPHKSG
2235

Q

QN

QTQQ

QKQL

GSGSPHSKAQIP
1906
GSGSPHARMQN
2016
GHDSPHKSG
2126
GSGSPHSKA
2236

P

QQ

QSKQ

QNQA

GSGSPHSKAQT
1907
GSGSPHVKSQN
2017
GHDSPHKTG
2127
GDDSPHKSG
2237

QL

QQ

QNQP

HNQQ

GSGSPHSKAQAP
1908
GQDSPHKSGQN
2018
GHDSPHKSG
2128
GHDSPHKSG
2238

S

QQ

QSPQ

QMIH

GHDSPHKSGRN
2239
GHDSPHKSVQN
2240
GHDSPHKSG
2241
TINGHDSPH
2728

HQ

RQ

QKMN

KSRLNQP

TINGSGSPHSKA
2242
TDRGSGSPHSK
2404
TINGSGSPHS
2566
TVDGHDSPH
2729

QNQQ

AQNQQ

KAQSTM

KSGQKQQ

TINGHDSPHKSG
2243
TINGSGSPHSKA
2405
TVNASGSPHS
2567
TINGQDSPH
2730

QNQQ

QIPP

KAQNQL

KSGQNQD

TIIGSGSPHSKAQ
2244
TVKGSGSPHSK
2406
TINGSGSPHS
2568
TIEGHDSPHK
2731

NRH

AQDQQ

KAQREM

SGRNQQ

TFPGSGSPHSKV
2245
NADGSGSPHSK
2407
TVHGSGSPHS
2569
TTNGHDSPH
2732

QNQQ

AQNQQ

KAQSQQ

KSGQNLL

TEKMSGSPHSK
2246
TDKVSGSPHSK
2408
TINGGGSPHS
2570
TINGHDSPH
2733

AQNQQ

AQNQQ

KSQNRQ

KSGQLVI

EINGRGSPHSKA
2247
TITGSGSPHSKA
2409
TINGSGSPHS
2571
TVNGHDSPH
2734

QNQQ

QTQL

KAQYRA

KSRQSQQ

TVNRNGSPHSK
2248
TINGSGSPHSKA
2410
TINGGGSPHS
2572
TINGHDSPH
2735

AQNQQ

QAPS

KAQRQQ

KSGRTQE

TVNGSGSPHSK
2249
NCVGSGSPHSK
2411
TEPMSGSPHS
2573
TINGHDSPH
2736

ARDQQ

AQNQQ

KAQNQQ

KSVQTHQ

TENGSGSPHSKA
2250
TIRDAGSPHSKA
2412
TINGSGSPHS
2574
TSNGHDSPH
2737

PNLQ

QNQQ

KNQWQQ

KSGQNQP

TEKTSGSPHSKA
2251
TVKDSGSPHSK
2413
ETAGSGSPHS
2575
VINGHDSPH
2738

QNQQ

AQNQQ

KAQNQQ

KSGQTQQ

TINGSGSPHSKA
2252
NALGSGSPHSK
2414
TINGSGSPHS
2576
TINGPDSPHK
2739

HVRQ

AQNQQ

KAQRMN

IGQNQQ

TVNGSGSPHSK
2253
VINGSGSPHSKG
2415
NNLGSGSPHS
2577
AVNGHDSPH
2740

APNQH

QNQQ

KAQNQQ

KSVQNQQ

TEKISGSPHSKA
2254
TVNGGGSPHSK
2416
TINGSGSPHA
2578
TINGHDSPH
2741

QNQQ

AQNQQ

KAQNHQ

KSRQDQH

TINGPGSPHSKA
2255
TIQDGGSPHSKA
2417
TIIKNGSPHS
2579
AINGPDSPHK
2742

HNQQ

QNQQ

KAQNQQ

SGQKQQ

TVNGSGSPHSKT
2256
TISGGGSPHSKA
2418
TINGSGSPHS
2580
TINGHDSPH
2743

QSQQ

QNQQ

KAGDSQ

KSRQSQH

SINESGSPHSKA
2257
TSNASGSPHSKA
2419
TINGSGSPHS
2581
TIYGHDSPH
2744

QNQQ

HNQQ

KLKSQQ

KSVQNQL

TERTSGSPHSKA
2258
TINGSGSPHSKA
2420
TINGSGSPHS
2582
TVNGHDSPH
2745

QNQQ

QNTY

KAQKIS

KSGQNLL

TINGSGSPHSKA
2259
TINGSGSPHSKS
2421
TEYNSGSPHS
2583
TENKSGSPHS
2746

QPAK

QNQH

KAQNQQ

KAQNQQ

TEKSSGSPHSKA
2260
TINGGGSPHSKA
2422
TINGSGSPHS
2584
TTNGQDSPH
2747

QNQQ

QDKQ

KAPSMQ

KSGQNQQ

TSYGNGSPHSK
2261
TEFVSGSPHSKA
2423
AINGSGSPHS
2585
TDKGSGSPH
2748

AQNQQ

QNQQ

KAQNQQ

SKAQNQQ

TEKGSGSPHSKA
2262
TVNGSGSPHSK
2424
TINGSGSPHS
2586
TIDGHDSPH
2749

QNQQ

AQNHL

KASPRQ

KSGRNQQ

TINGSGSPHSKS
2263
TREISGSPHSKA
2425
TINGSGSPHS
2587
TINGYDSPH
2750

QTQQ

QNQQ

KRMEQQ

KSGQYQH

TERISGSPHSKA
2264
TINGSGSPHSKA
2426
TINGSGSPHS
2588
TDNGHDSPH
2751

QNQQ

QIGM

KAQYQN

KSRQNQQ

TERASGSPHSKA
2265
TIDGSGSPHSKA
2427
TINGSGSPHS
2589
TINGHDSPH
2752

QNQQ

LNKQ

KAQYYV

KSWVRQQ

ELHGSGSPHSKA
2266
TIIGGGSPHSKA
2428
TINGSGSPHS
2590
TINGHESPHK
2753

QNQQ

QNPQ

KLRRQQ

SGQNQH

AINGSGSPHSKA
2267
QGEGSGSPHSK
2429
TINGSGSPHS
2591
TVNGHDSPH
2754

QNLA

AQNQQ

KAGCGQ

KIGHNQQ

TVNGSGSPHSKS
2268
TINGTGSPHSKA
2430
SMNGSGSPH
2592
TCNGHDSPH
2755

QNQL

PNQL

SRAQNQQ

KSGRNQQ

TERNSGSPHSKA
2269
TVNGSGSPHSK
2431
TINGSGSPHS
2593
TINGNGSPHS
2756

QNQQ

AQLQQ

KRLRQQ

KAQNHQ

SVNGNGSPHSK
2270
TFNGGGSPHSK
2432
TINGSGSPHS
2594
NVVGHDSPH
2757

AQNQQ

AQYQQ

LRRNQQ

KSGQNQQ

TFNGSGSPHSKA
2271
SINGSGSPHSKT
2433
TINGSGSPHS
2595
TINGHDSPH
2758

QGQQ

QSQQ

RGRNQQ

KSNAWQQ

TERVSGSPHSKA
2272
TVNGGGSPHSK
2434
TINGSGSPHS
2596
TDAGHDSPH
2759

QNQQ

AQHQQ

SRRNQQ

KSGQNQQ

TINGSGSPHSKA
2273
SEKGSGSPHSKA
2435
TINGSGSPHS
2597
TEVGHDSPH
2760

LNRQ

QNQQ

KAFRLQ

KSGQNQQ

TERLSGSPHSKA
2274
NVNGSGSPHSK
2436
TINGSCSPHR
2598
SELGHDSPH
2761

QNQQ

AQNQQ

KAQNQQ

KSGQNQQ

TDNGSGSPHSK
2275
GGEGSGSPHSK
2437
TINGSGSPHF
2599
TINGHDSPH
2762

AHNQQ

AQNQQ

LRQNQQ

KSGQSVP

TFHGSGSPHSKT
2276
TINGSGSPHSKA
2438
TINGSGSPHS
2600
TINGHESPHK
2763

QNQQ

QRMS

LRFNQQ

SGQNIQ

TINGGGSPHSKA
2277
TINGSGSPHSKA
2439
TINGSGSPHS
2601
TINGHDSPH
2764

QTQQ

QGIL

KWLLQQ

KSVQNHL

TSNGSGSPHSKA
2278
EFVGSGSPHSKA
2440
TINGSGSPHS
2602
TINGHDSPH
2765

QNPP

QNQQ

KRRLQQ

KIGLDQQ

TINGSGSPHSKA
2279
TIIGSGSPHSKAQ
2441
TINGSGSPHS
2603
TSNASGSPHS
2766

QNLQ

DRQ

KAQRKL

KAQHQQ

TVHGNGSPHSK
2280
SDKGSGSPHSK
2442
TINGSGSPHS
2604
TINGHDSPH
2767

AQNQQ

AQNQQ

KALRRQ

KRGPDQQ

TINGGGSPHSKA
2281
TEQVSGSPHSKA
2443
TINGSGSPHS
2605
TINGMGSPH
2768

QNQQ

QNQQ

KAQRLR

SKTQNQQ

TENMSGSPHSK
2282
TEHVSGSPHSKA
2444
YLSGSGSPHS
2606
TIKGHDSPH
2769

AQNQQ

QNQQ

KAQNQQ

KSGESQQ

TENVSGSPHSKA
2283
TINGSGSPHSKA
2445
TINGSGSPHS
2607
TINGHDSPH
2770

QNQQ

RDWQ

KAQRRL

KHGQNHQ

TSSGSGSPHSKA
2284
TENASGSPHSKA
2446
TINGSGSPHS
2608
TVNGTGSPH
2771

QYQQ

QNQQ

KARRQQ

SKAQNQL

TIDGGGSPHSKA
2285
EVQGSGSPHSK
2447
TINGSGSPHS
2609
TIIGHDSPHK
2772

QNKQ

AQNQQ

KARRLQ

SGQYQH

TEKVSGSPHSKA
2286
TINGSGSPHSKA
2448
TINGSGSPHS
2610
TSNGHDSPH
2773

QNQQ

QNTH

KSRRQQ

KSVQNKQ

AINGSGSPHSKA
2287
TINGSGSPHSKA
2449
TINGLLSPHW
2611
IVNGQVSPH
2774

QDQE

PNLQ

KAQNQQ

KSGQNQQ

TCNKSGSPHSKA
2288
TINGSGSPHSKA
2450
TINGSGSPHS
2612
TVNGHDSPH
2775

QNQQ

QERS

KARLRQ

KSGQRQL

TINGGGSPHSKA
2289
TSNGSGSPHSKA
2451
TINGSGSPHS
2613
TVNGHDSPH
2776

QNQL

QNYQ

KASKRQ

KIGQNQL

NINGGGSPHSKA
2290
TEYISGSPHSKA
2452
TINGSGSPHV
2614
TINGHDSPH
2777

QNQQ

QNQQ

RRQNQQ

KSGQIIV

TEHLSGSPHSKA
2291
TINGSGSPHSKA
2453
TINGSGSPHS
2615
IGNGHESPH
2778

QNQQ

QRTC

KAQLYR

KSGQNQQ

AEMGSGSPHSK
2292
TINGSGSPHSKA
2454
GLSGSGSPHS
2616
EVMGHDSPH
2779

AQNQQ

QIGH

KAQNQQ

KSGQNQQ

ATNGSGSPHSK
2293
NCWGSGSPHSK
2455
TINGSGSPHS
2617
TINGYDSPH
2780

AQNHQ

AQNQQ

KAQLTV

KSGQKQS

AIKGSGSPHSKA
2294
TINGSGSPHSKA
2456
TINGHDSPHK
2618
TIHGNGSPHS
2781

QDQQ

QGAI

RGQHRQ

KAQNQE

TINGGGSPHSKS
2295
TDVNSGSPHSK
2457
MPEGHDSPH
2619
YQVGHDSPH
2782

QNQL

AQNQQ

KSGQNQQ

KSGQNQQ

TVNGNGSPHSK
2296
SDIGSGSPHSKA
2458
MEGGHDSPH
2620
TIKGDDSPH
2783

AQNKQ

QNQQ

KSGQNQQ

KSVQNQQ

TINGSGSPHSKG
2297
TINGSGSPHSKA
2459
MEYGHDSPH
2621
TINGHDSPH
2784

HWQQ

QVPP

KSGQNQQ

KSVQSHQ

TDKTSGSPHSKA
2298
TINGSGSPHSKA
2460
AEWGHDSPH
2622
TINGHDSPH
2785

QNQQ

QVQQ

KSGQNQQ

KSGQFVV

TFKGSGSPHSKA
2299
TINGSGSPHSKA
2461
CEWGHDSPH
2623
TVNGHDSPH
2786

PNQQ

LMRQ

KSGQNQQ

KSRQNLQ

TVNGSGSPHSK
2300
TINGSGSPHSKA
2462
ANNGQDSPH
2624
ATNGHNSPH
2787

AQNQL

QYSV

KSGQNQQ

KSGQNQE

TINGSGSPHSKR
2301
NSIGSGSPHSKA
2463
IPEGHDSPHK
2625
AINGHDSPH
2788

PEQQ

QNQQ

SGQNQQ

KSAQNQQ

TINGSGSPHSKA
2302
TINGSGSPHSKV
2464
ADMGHDSPH
2626
TEHGHDSPH
2789

QRTM

PNLQ

KSGQNQQ

KSGQNQQ

TEKASGSPHSKA
2303
AINGSGSPHSKA
2465
IEYGHDSPHK
2627
TIYGHDSPH
2790

QNQQ

QSQQ

SGQNQQ

KSGQSQP

SDQGSGSPHSKA
2304
TINGSGSPHSKA
2466
ADYGHDSPH
2628
TISGHESPHK
2791

QNQQ

QAIT

KSGQNQQ

SGQNEQ

TEITSGSPHSKA
2305
TINGSGSPHSKA
2467
IETGHDSPHK
2629
AIIGHDSPHK
2792

QNQQ

QKTL

SGQNQQ

SAQNQQ

TDKSSGSPHSKA
2306
TVNGSGSPHSK
2468
MEWGHDSPH
2630
AIDGHDSPH
2793

QNQQ

AQNQW

KSGQNQQ

KSGQNQL

TIDGSGSPHSKA
2307
TINGSGSPHSKA
2469
CEYGHDSPH
2631
TIMGHDSPH
2794

QNQQ

QLHH

KSGQNQQ

KSVQNQQ

TVNGNGSPHSK
2308
TEQTSGSPHSKA
2470
RINGHDSPHK
2632
EVGGHDSPH
2795

AQNQH

QNQQ

SGQKQQ

KSGQNQQ

NTNGSGSPHSK
2309
TINGSGSPHSKA
2471
MEIGHDSPH
2633
TINGHDSPH
2796

AQNQQ

QNII

KSGQNQQ

KSAQNLL

TETHSGSPHSKA
2310
NSLGSGSPHSKA
2472
LEYGHDSPH
2634
TINASGSPHS
2797

QNQQ

QNQQ

KSGQNQQ

KAINQQ

TINGGGSPHSKA
2311
TIPMEGSPHSKA
2473
ADWGHDSPH
2635
AINGNGSPH
2798

LNQQ

QNQQ

KSGQNQQ

KRGQNQQ

TINGSGSPHSKA
2312
TINGSGSPHSKA
2474
IEIGHDSPHK
2636
SEMGHDSPH
2799

LHQH

QGHH

SGQNQQ

KSGQNQQ

TINGTGSPHSKA
2313
TDRTSGSPHSKA
2475
DIMGHDSPH
2637
AQQGHDSPH
2800

QNHQ

QNQQ

KSGQNQQ

KSGQNQQ

TINGSGSPHSKA
2314
TINGSGSPHSKA
2476
FEQGHDSPH
2638
AINGHDSPH
2801

QHRI

QSKV

KSGQNQQ

KSGQSLQ

TINGSGSPHSKA
2315
EVVGSGSPHSK
2477
MEFGHDSPH
2639
TINGSGSPHS
2802

QYIH

AQNQQ

KSGQNQQ

KAPNQQ

TENISGSPHSKA
2316
TINGSGSPHSKA
2478
CDQGHDSPH
2640
CGEGHDSPH
2803

QNQQ

QLPS

KSGQNQQ

KSGQNQQ

TIIGGGSPHSKA
2317
TINGSGSPHSKAI
2479
LPEGHDSPH
2641
TVNGHDSPH
2804

HNQQ

GKQ

KSGQNQQ

KSAQNHQ

TINGSGSPHSKA
2318
TEPTSGSPHSKA
2480
IENGHDSPHK
2642
TVNGHDSPH
2805

QKFE

QNQQ

SGQNQQ

KSGQTQL

TSNESGSPHSKA
2319
TVNGGGSPHSK
2481
MESGHDSPH
2643
TNNGHDSPH
2806

QNHQ

SQNQQ

KSGQNQQ

KSGRNRQ

TINGSGSPHSKA
2320
TINGSGSPHSKA
2482
AEIGHDSPHK
2644
TINGHDSPH
2807

QFPS

QAIH

SGQNQQ

KYGQNEQ

TERPSGSPHSKA
2321
TINGSGSPHSKA
2483
VEYGHDSPH
2645
TINGNGSPHS
2808

QNQQ

QHGL

KSGQNQQ

KAPNLQ

TINGNGSPHSKA
2322
SELGSGSPHSKA
2484
IINGHDSPHK
2646
SINGHDSPHK
2809

QNPL

QNQQ

SGLTQQ

SQQNQQ

SIKGNGSPHSKA
2323
TINGSGSPHSKA
2485
TSNGDDSPH
2647
TIGGHDSPH
2810

QNQQ

QFMC

KSGRNQQ

KSGQNQQ

TERMSGSPHSK
2324
TINVSGSPHSKA
2486
IEVGHDSPHK
2648
TINGHDSPH
2811

AQNQQ

QGQQ

SGQNQQ

KSVQSKQ

TERSSGSPHSKA
2325
TINGGGSPHSKA
2487
MEMGHDSPH
2649
ELVGHDSPH
2812

QNQQ

QNQM

KSGQNQQ

KSGQNQQ

TELHSGSPHSKA
2326
TVNGSGSPHSK
2488
AEVGHDSPH
2650
ELMGHDSPH
2813

QNQQ

AQHLQ

KSGQNQQ

KSGQNQQ

TELTSGSPHSKA
2327
TIRENGSPHSKA
2489
MDAGHDSPH
2651
TINGNDSPH
2814

QNQQ

QNQQ

KSGQNQQ

KIGHNQQ

TINGSGSPHSKA
2328
TINGSGSPHSKT
2490
VEWGHDSPH
2652
TIKGGGSPHS
2815

HNQQ

QNHQ

KSGQNQQ

KAQDQQ

TINGGGSPHSKA
2329
TINGSGSPHSKA
2491
AEQGHDSPH
2653
TVNGHDSPH
2816

QSQQ

QPAR

KSGQNQQ

KSGQTQQ

TINGSGSPHSKA
2330
TVNGSGSPHSK
2492
LEWGHDSPH
2654
TINGQDSPH
2817

QAIK

AQSLQ

KSGQNQQ

KSGQNPL

TENTSGSPHSKA
2331
TINGSGSPHSKS
2493
MELGHDSPH
2655
TVNASGSPH
2818

QNQQ

QSQL

KSGQNQQ

SKAQNHQ

TIDGSGSPHSKG
2332
TINGSASPHSKA
2494
METGHDSPH
2656
TINGHDSPH
2819

QNRQ

HSQQ

KSGQNQQ

KSGRDQK

NINGSGSPHSKA
2333
TWQNSGSPHSK
2495
MEAGHDSPH
2657
TINGHDSPH
2820

QSQQ

AQNQQ

KSGQNQQ

KSVHNQQ

TINGSVSPHGKA
2334
TINGSGSPHSKA
2496
IESGHDSPHK
2658
TINGHDSPH
2821

QNQL

QDRQ

SGQNQQ

KSGQWKR

TSNASGSPHSKA
2335
TINGSGSPHSKA
2497
MEVGHDSPH
2659
TIDGSGSPHS
2822

QNQL

QMPS

KSGQNQQ

KAENRQ

TEARSGSPHSKA
2336
TNNGGGSPHSK
2498
CEIGHDSPHK
2660
NEIGHDSPH
2823

QNQQ

AQNLQ

SGQNQQ

KSGQNQQ

TEKNSGSPHSKA
2337
TINGSGSPHSKA
2499
ATNGHDSPH
2661
AINGHDSPH
2824

QNQQ

QGSL

KSGLNQQ

KSGQSQQ

TANGSGSPHSK
2338
TEVTSGSPHSKA
2500
MDGGHDSPH
2662
IINGHDSPHK
2825

AQYQQ

QNQQ

KSGQNQQ

SRQAQQ

TVNGSGSPHSK
2339
SINGGGSPHSKA
2501
QEVGHDSPH
2663
TPNGHDSPH
2826

AQYQH

QYQQ

KSGQNQQ

KSGQNQQ

TINGSGSPHTKA
2340
TVIGSGSPHSKS
2502
ADQGHDSPH
2664
ITNGHDSPH
2827

QNPQ

QNQQ

KSGQNQQ

KSGQTQQ

TINGSGSPHSKG
2341
AVNVSGSPHSK
2503
TINGHESPHK
2665
TINGHDSPH
2828

QNPP

AQNQQ

SAQNHQ

KSVQNHQ

TIIGSGSPHSKAQ
2342
TVNGNGSPHSK
2504
TINGHDSPHK
2666
TINGHDSPH
2829

HQL

SQNQQ

SAQNQW

KSKQNQQ

TINGSGSPHSKA
2343
TDRNSGSPHSK
2505
NMNGHDSPH
2667
TINGHDSPH
2830

QSPP

AQNQQ

KSGQNTH

KSAQNQL

TIYGSGSPHSKA
2344
TINGSGSPHSKA
2506
IEMGHDSPH
2668
TVNGHDSPH
2831

QNQQ

QVPA

KSGQNQQ

KSGQTQP

TINGSGSPHSKA
2345
GVLGSGSPHSK
2507
TINGHDSPHK
2669
TDQGHDSPH
2832

QAKL

AQNQQ

SGRRRQ

KSGQNQQ

TDKNSGSPHSK
2346
TLNGNGSPHSK
2508
ISNGHDSPHK
2670
TINGHDSPH
2833

AQNQQ

AQNLQ

SAQNQQ

KLWINQQ

TINGSGSPHSKT
2347
AINGSGSPHSKA
2509
TGNGHDSPH
2671
GINGPDSPHK
2834

KSQQ

QDKQ

KSGQYQQ

SGQNQQ

TINGSGSPHSKA
2348
TSNGSGSPHSKA
2510
TINGNYSPHK
2672
SEIGHDSPHK
2835

QDRP

HYQQ

IGQNQQ

SGQNQQ

TINGIGSPHSKA
2349
TINGSGSPHSKA
2511
TINGHDSPHK
2673
TINGHDSPH
2836

QNLG

QVPS

SRQNDQ

KSVQKQL

TINGSGSPHSKA
2350
TELRSGSPHSKA
2512
QQQGHDSPH
2674
TINGHPSPH
2837

QSQQ

QNQQ

KSGQNQQ

WKGQNQQ

TENLSGSPHSKA
2351
NINGSGSPHSKA
2513
HDWGHDSPH
2675
TVNGHDSPH
2838

QNQQ

QNHQ

KSGQNQQ

KSGRNQL

TINGSGSPHSKA
2352
TVNGGGSPHSK
2514
IEGGHDSPHK
2676
TIKGSGSPHS
2839

QAFH

AQNHQ

SGQNQQ

KVQDQQ

TINGSGSPHSKA
2353
TINGSGSPHSKA
2515
TFNRSGSPHS
2677
SEKGHDSPH
2840

QKQQ

RGEQ

KAQNQQ

KSGQNQQ

TFSGSGSPHSKA
2354
TINGGGSPHSKA
2516
AINGHDSPH
2678
WSAGHDSPH
2841

QNLQ

QYQH

KSGQIRQ

KSGQNQQ

AINGSGSPHSKA
2355
TEDLSGSPHSKA
2517
TINGHDSPHK
2679
ELAGHDSPH
2842

QNAQ

QNQQ

IGQNQH

KSGQNQQ

TESWSGSPHSKA
2356
TINGSGSPHSKA
2518
AINGYDSPH
2680
TINGHDSPH
2843

QNQQ

PGQQ

KSGQKQQ

KMGRNQQ

TTNGSGSPHSKA
2357
TIPKNGSPHSKA
2519
TESGHDSPH
2681
TINGHDSPH
2844

HNQL

QNQQ

KSGQNQQ

KSGISIQ

TVNGNGSPHSK
2358
TINGSGSPHSKA
2520
TINGHDSPHK
2682
TSNGHDSPH
2845

AQNHQ

QSLQ

SGQSVQ

KSVQNLQ

TEDKSGSPHSKA
2359
TINGSGSPHSKR
2521
TINGHESPHK
2683
QTQGHDSPH
2846

QNQQ

LEQQ

SGRSQQ

KSGQNQQ

TESASGSPHSKA
2360
TERGSGSPHSKA
2522
TINGHDSPHK
2684
TINGHDSPH
2847

QNQQ

QNQQ

SGQNKL

KMAHNQQ

TNNGSGSPHSK
2361
TVNGSGSPHSK
2523
TINGHDSPHK
2685
AINGSGSPHS
2848

AQNQQ

APNQQ

TGQNQQ

KAQTQQ

TSNGGGSPHSK
2362
TSNGSGSPHSKA
2524
TINGRGSPHK
2686
TINGHDSPH
2849

AQNLQ

QNQS

RGQNQQ

KHGQNQQ

TDKMSGSPHSK
2363
TINGSGSPHSKA
2525
TINGSGSPHT
2687
GADGHDSPH
2850

AQNQQ

QKVI

KAQNPP

KSGQNQQ

EVHGSGSPHSK
2364
TEGISGSPHSKA
2526
TINGQDSPHK
2688
VGEGHDSPH
2851

AQNQQ

QNQQ

SGQHQQ

KSGQNQQ

TINGSGSPHSKA
2365
TINGSGSPHSKA
2527
SINGHDSPHK
2689
ANEGHDSPH
2852

QKLN

QNND

SGQIQH

KSGQNQQ

TINGGGSPHSKS
2366
TINGSGSPHSKA
2528
AINGHDSPH
2690
TEAKSGSPHS
2853

QNQH

QSVH

KSGPRQQ

KAQNQQ

TVNGGGSPHSK
2367
TINGSGSPHSKA
2529
TVNGHDSPH
2691
TINGHDSPH
2854

AQSQQ

QPLG

KSGHTQQ

KSVQSQQ

TTNGSGSPHSKA
2368
TINKEGSPHSKA
2530
SINGHDSPHK
2692
TIPGSGSPHS
2855

QYQH

QNQQ

SGQRQH

KAQNLQ

TISGSGSPHSKA
2369
TCNASGSPHSK
2531
SLNGSGSPHT
2693
TINGHDSPH
2856

QYQH

AQNQQ

KAQNQQ

KSGQTVC

TESTSGSPHSKA
2370
AINGSGSPHSKA
2532
AINGHDSPH
2694
ELRGHDSPH
2857

QNQQ

HNQE

KSAQSQQ

KSGQNQQ

TINGSGSPHSKS
2371
TEGLSGSPHSKA
2533
SIYGHESPHK
2695
CQIGHDSPH
2858

QNVQ

QNQQ

SGQNQQ

KSGQNQQ

SINGSGSPHSKA
2372
TRDASGSPHSK
2534
TVNGHDSPH
2696
GVMGHDSPH
2859

QAQQ

AQNQQ

KSLQNQQ

KSGQNQQ

TVNGSGSPHSK
2373
TSNGSGSPHSKA
2535
TINGHGSPHS
2697
ACDGHDSPH
2860

AQNLQ

QNLQ

KAQNPQ

KSGQNQQ

TVRDSGSPHSKA
2374
TGNGSGSPHSK
2536
TSNGYDSPH
2698
TINGQDSPH
2861

QNQQ

AQIQQ

KSGQKQQ

KSGQYQQ

TFNASGSPHSKA
2375
TVNGGGSPHSK
2537
TVNGHDSPH
2699
TINGHDSPH
2862

PNQQ

AQNLQ

KSGRNQE

KSGQQIM

TDRMSGSPHSK
2376
TDRSSGSPHSKA
2538
TTNGHDSPH
2700
TINGHDSPH
2863

AQNQQ

QNQQ

KSGQTQL

KSRQNEQ

TINGSGSPHSKA
2377
TINGSGSPHSKA
2539
AINGHDSPH
2701
ASNGHDSPH
2864

QTPP

QVRN

KSEKNQQ

KSGLNHQ

TIKGSGSPHSKA
2378
TINGSGSPHSKA
2540
IINGRDSPHK
2702
TVNGHDSPH
2865

QNQQ

PSNQ

SGQDQQ

KSGQSQP

NHIGSGSPHSKA
2379
TINGSGSPHSKA
2541
TISGHDSPHK
2703
NELGHDSPH
2866

QNQQ

QVGH

TGHNQQ

KSGQNQQ

TINGSGSPHSKA
2380
NAIGSGSPHSKA
2542
SINGYDSPHK
2704
AAEGHDSPH
2867

QYQH

QNQQ

SGQTQQ

KSGQNQQ

TIPIDGSPHSKAQ
2381
AENGSGSPHSK
2543
TINGHESPHK
2705
GQNGHDSPH
2868

NQQ

AQNQQ

SGQTQQ

KSGQNQQ

TINGSGSPHSKA
2382
TINGSGSPHSKA
2544
TINGHDSPHK
2706
NEFGHDSPH
2869

QGQQ

QRDI

SGQSKQ

KSGQNQQ

TFNGSGSPHNK
2383
TINGSGSPHSKA
2545
AIIGHESPHK
2707
TSIGYDSPHK
2870

AQNHQ

QMPN

SGQNQQ

SGQNQQ

ESDGSGSPHSKA
2384
TVNGSGSPHSKS
2546
TINGHDSPHK
2708
TDNGHDSPH
2871

QNQQ

QNQQ

TGQNQP

KSGQNLQ

TINGSGSPHSKA
2385
TIPAIGSPHSKAQ
2547
AINGHDSPH
2709
TITGHDSPHK
2872

QPPA

NQQ

KSGQSPQ

SRQDQQ

TINGSGSPHSKA
2386
TINGSGSPHSKA
2548
TIKGNDSPHK
2710
AEHGHDSPH
2873

QERP

RGLQ

SVQNQQ

KSGQNQQ

TIKGSGSPHSKA
2387
TELGSGSPHSKA
2549
TEFGHDSPH
2711
TINGDDSPH
2874

QDLQ

QNQQ

KSGQNQQ

KSGQKQL

TDLKSGSPHSKA
2388
AETGSGSPHSKA
2550
TINGHDSPHK
2712
EILGHDSPHK
2875

QNQQ

QNQQ

SAQNYQ

SGQNQQ

TINGGGSPHSKA
2389
TINGSGSPHSKL
2551
TFNGSASPHS
2713
TIHGSGSPHS
2876

QNPP

QKQQ

KALNQQ

KAQNQA

TINGSGSPHSKA
2390
TINGSGSPHSKA
2552
TINGHESPHK
2714
AINGDDSPH
2877

QAMH

PSLQ

SAQNRQ

KSGHNQQ

TVPNSGSPHSKA
2391
TINGSGSPHSKA
2553
TTNGHDSPH
2715
TSNGHNSPH
2878

QNQQ

QRDQ

KSGQNQQ

KSGQNQE

TVIGSGSPHSKA
2392
TDVGSGSPHSK
2554
TIKGQDSPHK
2716
TINGHDSPH
2879

LNQQ

AQNQQ

IGQNQQ

KSGQMIH

TINGSGSPHSKA
2393
TINGSGSPHSKN
2555
TVNGHDSPH
2717
NAIGHDSPH
2880

QHPS

RDQQ

KSGQNHL

KSGQNQQ

TINGLGSPHSKS
2394
SINGSGSPHSKA
2556
SINGHDSPHK
2718
VINGHDSPH
2881

QNQQ

PNLQ

SGQYQH

KSGRNHQ

TINGTGSPHSKA
2395
TINGSGSPHSKA
2557
TINGNDSPHK
2719
TITGHDSPHK
2882

QNQQ

QAKG

SVQNHQ

SVQNRQ

TINGSGSPHSKA
2396
TVNGSGSPHSK
2558
TITGHDSPHK
2720
TINGHDSPH
2883

PGLQ

AQDKQ

SGQNQW

KSGQKMN

TINGSGSPHSKA
2397
TINGGGSPHSKA
2559
TNNGHDSPH
2721
TIHGHDSPH
2884

QGIR

QNPQ

KSVQNQH

KSGQSQQ

TESHSGSPHSKA
2398
TINGSGSPHSKA
2560
TIDGHDSPHK
2722
TEIGHDSPHK
2885

QNQQ

QSAH

SGQNQH

SGQNQQ

TINGSGSPHSKA
2399
TINGNGSPHSKS
2561
TVNGHDSPH
2723
TINGHDSPH
2886

QAPA

QNQH

KSGQTRQ

KSGQYQH

TINGSGSPHSKS
2400
TVPTSGSPHSKA
2562
TVNGHDSPH
2724
NCLGSGSPH
2403

QSQQ

QNQQ

KSGQNLH

SKAQNQQ

AEHGSGSPHSK
2401
TIDGSGSPHSKS
2563
AISGHDSPHK
2725
AINRSGSPHS
2565

AQNQQ

QNHQ

SGLNQQ

KAQDQQ

TEDRSGSPHSKA
2402
TDVKSGSPHSK
2564
AINGHDSPH
2726
TITGHDSPHK
2727

QNQQ

AQNQQ

KSAQNQE

SGQHLQ

GSGSPHSKAQN
2887
GSGSPHSKAQS
2936
GSGSPHSKA
2985
GSGSPHSKA
3034

RHT

QQT

QRMST

QNNDQ

GSGSPHSKVQN
2888
ASGSPHSKAQN
2937
GSGSPHSKA
2986
GSGSPHSKA
3035

QQT

QLT

QGILT

QSVHT

MSGSPHSKAQN
2889
RSGSPHSKAQN
2938
GSGSPHSKA
2987
GSGSPHSKA
3036

QQT

QQT

QDRQT

QPLGT

GSGSPHSKARD
2890
GSGSPHSKAQY
2939
GSGSPHSKA
2988
GSGSPHSKA
3037

QQT

QHT

RDWQT

HNQET

GSGSPHSKAPNL
2891
GSGSPHTKAQN
2940
GSGSPHSKA
2989
GSGSPHSKA
3038

QT

PQS

QNTHD

QNLQI

TSGSPHSKAQN
2892
GSGSPHSKGQN
2941
GSGSPHSKAP
2990
GSGSPHSKA
3039

QQT

PPT

NLQI

QIQQT

GSGSPHSKAHV
2893
GSGSPHSKAQH
2942
GSGSPHSKA
2991
GSGSPHSKA
3040

RQT

QLT

QERST

QVRNT

GSGSPHSKAPNQ
2894
GSGSPHSKAQSP
2943
GSGSPHSKA
2992
GSGSPHSKA
3041

HT

PT

QNYQT

PSNQT

ISGSPHSKAQNQ
2895
GSGSPHSKAQA
2944
GSGSPHSKA
2993
GSGSPHSKA
3042

QT

KLT

QRTCT

QVGHT

GSGSPHSKTQSQ
2896
GSGSPHSKTKSQ
2945
GSGSPHSKA
2994
GSGSPHSKA
3043

QT

QT

QIGHT

QRDIT

GSGSPHSKAQN
3032
GSGSPHSKAQD
2946
GSGSPHSKA
2995
GSGSPHSKA
3044

QST

RPT

QGAIT

QMPNT

ESGSPHSKAQN
2898
GSGSPHSKAQS
2947
GSGSPHSKA
2996
GSGSPHSKA
3045

QQI

QQL

QVPPT

RGLQT

GSGSPHSKAQPA
2899
GSGSPHSKAQA
2948
GSGSPHSKA
2997
GSGSPHSKL
3046

KT

FHT

QVQQI

QKQQT

SSGSPHSKAQNQ
2900
GSGSPHSKAQK
2949
GSGSPHSKAL
2998
GSGSPHSKA
3047

QT

QQD

MRQT

PSLQT

GSGSPHSKSQTQ
2901
GSGSPHSKAQN
2950
GSGSPHSKA
2999
GSGSPHSKA
3048

QN

AQT

QYSVT

QRDQT

ASGSPHSKAQN
2902
WSGSPHSKAQN
2951
GSGSPHSKVP
3000
GSGSPHSKN
3049

QQT

QQT

NLQT

RDQQT

GSGSPHSKAQN
2903
GSGSPHSKAHN
2952
GSGSPHSKA
3001
GSGSPHSKA
3050

LAT

QLT

QSQQI

QAKGT

GSGSPHSKSQNQ
2904
GSGSPHSKAQN
2953
GSGSPHSKA
3002
GSGSPHSKA
3051

LT

QQY

QAITT

QSAHT

NSGSPHSKAQN
2905
GSGSPHSKAQK
2954
GSGSPHSKA
3003
GSGSPHSKS
3052

QQT

LNT

QKTLT

QNHQT

GSGSPHSKAQG
2906
GSGSPHSKSQN
2955
GSGSPHSKA
3004
RSGSPHSKA
3053

QQT

VQT

QNQWT

QDQQT

VSGSPHSKAQN
2907
GSGSPHSKAQA
2956
GSGSPHSKA
3005
GSGSPHSKA
3054

QQT

QQT

QLHHT

QSTMT

GSGSPHSKALNR
2908
GSGSPHSKAQN
2957
GSGSPHSKA
3006
GSGSPHSKA
3055

QS

LQA

QNIII

QREMT

LSGSPHSKAQN
2909
DSGSPHSKAQN
2958
GSGSPHSKA
3007
GSGSPHSKA
3056

QQT

QQT

QGHHT

QYRAT

GSGSPHSKAHN
2910
ASGSPHSKAPN
2959
GSGSPHSKA
3008
GSGSPHSKN
3057

QQT

QQT

QSKVT

QWQQT

GSGSPHSKTQN
2911
GSGSPHSKAQTP
2960
GSGSPHSKA
3009
GSGSPHSKA
3058

QQT

PT

QLPST

QRMNT

GSGSPHSKAQNP
2912
GSGSPHSKAQY
2961
GSGSPHSKAI
3010
GSGSPHAKA
3059

PT

QHA

GKQT

QNHQT

GSGSPHSKAQN
2913
GSGSPHSKAQG
2962
GSGSPHSKA
3011
GSGSPHSKA
3060

LQT

QQA

QAIHT

GDSQT

GSGSPHSKAQY
2914
GSGSPHNKAQN
2963
GSGSPHSKA
3012
GSGSPHSKL
3061

QQT

HQT

QHGLT

KSQQT

GSGSPHSKAQD
2915
GSGSPHSKAQPP
2964
GSGSPHSKA
3013
GSGSPHSKA
3062

QET

AT

QFMCT

QKIST

KSGSPHSKAQN
2916
GSGSPHSKAQE
2965
VSGSPHSKA
3014
GSGSPHSKA
3063

QQT

RPT

QGQQT

PSMQT

GSGSPHSKAQN
2917
GSGSPHSKAQD
2966
GSGSPHSKA
3015
GSGSPHSKA
3064

HQT

LQT

QHLQT

SPRQT

GSGSPHSKAQD
2918
GSGSPHSKAQA
2967
GSGSPHSKTQ
3016
GSGSPHSKR
3065

QQT

MHT

NH?N

MEQQT

GSGSPHSKGHW
2919
GSGSPHSKALN
2968
GSGSPHSKA
3017
GSGSPHSKA
3066

QQT

QQT

QPART

QYQNT

GSGSPHSKAPNQ
2920
GSGSPHSKAQH
2969
GSGSPHSKA
3018
RSGSPHSKA
3067

QT

PST

QSLQT

QNQQI

GSGSPHSKAQN
2921
GSGSPHSKAPGL
2970
GSGSPHSKSQ
3019
GSGSPHTKA
3068

QLI

QT

SQLT

QNPPT

GSGSPHSKRPEQ
2922
GSGSPHSKAQGI
2971
GSGSPHSKA
3020
GSGSPHTKA
3069

QT

RT

QDRQS

QNQQT

GSGSPHSKAQRT
2923
GSGSPHSKAQA
2972
GSGSPHSKA
3021
ASGSPHSKA
3070

MT

PAT

QMPST

QHQQT

GSGSPHSKAQN
2924
GSGSPHSKSQSQ
2973
GSGSPHSKA
3022
ASGSPHSKAI
3071

QQH

QI

QGSLT

NQQT

HSGSPHSKAQN
2925
GSGSPHSKAQIP
2974
GSGSPHSKSQ
3023
GSGSPHSKA
3072

QQT

PT

NQQT

PNQQH

GSGSPHSKALH
2926
GSGSPHSKAQT
2975
GSGSPHSKA
3024
ASGSPHSKA
3073

QHT

QLT

QVPAT

QNHQT

GSGSPHSKAQH
2927
GSGSPHSKAQA
2976
GSGSPHSKA
3025
GSGSPHSKA
3074

RIT

PST

QDKQT

ENRQT

GSGSPHSKAQYI
2928
GSGSPHSKGQN
2977
GSGSPHSKA
3026
GSGSPHSKV
3075

HT

QQT

HYQQT

QDQQT

GSGSPHSKAQKF
2929
ASGSPHSKAHN
2978
GSGSPHSKA
3027
GSGSPHSKA
3076

ET

QQT

QVPST

QTQQA

ESGSPHSKAQN
2930
GSGSPHSKAQN
2979
GSGSPHSKA
3028
GSGSPHSKA
3077

HQT

TYA

RGEQT

QNQAT

GSGSPHSKAQFP
2931
GSGSPHSKSQN
2980
GSGSPHSKAP
3029
GSGSPHSKG
2935

ST

QHI

GQQT

QNRQT

PSGSPHSKAQNQ
2932
GSGSPHSKAQN
2981
GSGSPHSKA
3030
GSGSPHSKA
2984

QT

HLT

QSLQI

QLQQT

GSGSPHSKAHN
2933
GSGSPHSKAQIG
2982
GSGSPHSKRL
3031
GSGSPHSKA
3033

QQR

MT

EQQT

QKVIT

GSGSPHSKAQAI
2934
GSGSPHSKALN
2983
GHDSPHKHG
3160
GHDSPHKSG
3202

KT

KQT

QNHQT

QWKRT

GHDSPHKSGQIR
3078
GHDSPHKSGQN
3119
LHDSPHKSG
3161
GHDSPHKSG
3203

QT

QHA

QNQQT

QSQQI

GHDSPHKIGQN
3079
GHDSPHKSGQT
3120
GHDSPHKSV
3162
GHDSPHKSR
3204

QHA

RQT

QNKQT

QAQQT

GYDSPHKSGQK
3080
GHDSPHKSGQN
3121
GQVSPHKSG
3163
GHDSPHKSV
3205

QQT

LHT

QNQQT

QNHQI

GHDSPHKSGQN
3081
GHDSPHKSGLN
3122
GHDSPHKSG
3164
GHDSPHKSK
3206

QQT

QQT

QRQLT

QNQQA

GHDSPHKSGQS
3082
GHDSPHKSAQN
3123
GHDSPHKIG
3165
GHDSPHKSA
3207

VQT

QET

QNQLT

QNQLN

GHESPHKSGRSQ
3083
GHDSPHKSGQH
3124
GHDSPHKSG
3166
GHDSPHKSG
3208

QT

LQT

QIIVT

QTQPT

GHDSPHKSGQN
3084
GHDSPHKSRLN
3125
IHDSPHKSGQ
3167
FHDSPHKSG
3209

KLE

QPT

NQQT

QNQQT

GHDSPHKTGQN
3085
GHDSPHKSGQK
3126
GYDSPHKSG
3168
GHDSPHKLW
3210

QQK

QQT

QKQST

INQQT

GRGSPHKRGQN
3086
GQDSPHKSGQN
3127
MHDSPHKSG
3169
GPDSPHKSG
3211

QQT

QDT

QNQQT

QNQQT

GHDNPHKSGQN
3087
GHDSPHKSGRN
3128
GDDSPHKSV
3170
GHDSPHKSV
3212

QQT

QQT

QNQQT

QKQLT

GQDSPHKSGQH
3088
GHDSPHKSGQN
3129
GHDSPHKSV
3171
GHDSPHKSG
3213

QQA

LLT

QSHQT

RNQLA

GHDSPHKSGQIQ
3089
GHDSPHKSGQL
3130
GHDSPHKSG
3172
VHDSPHKSG
3214

HT

VIT

QFVVT

QNQQS

GHDSPHKSGPR
3090
GHDSPHKSRQS
3131
GHDSPHKSR
3173
GHDSPHKMG
3215

QQT

QQT

QNLQT

RNQQS

GHDSPHKSGHT
3091
GHDSPHKSGRT
3132
GHNSPHKSG
3174
GHDSPHKSGI
3216

QQT

QET

QNQET

SIQT

GHDSPHKSGQR
3092
GHDSPHKSVQT
3133
GHDSPHKSA
3175
VHDSPHKSG
3217

QHT

HQT

QNQQI

QNQQT

GHDSPHKSAQS
3093
GHDSPHKSGQN
3134
NHDSPHKSG
3176
GHDSPHKSV
3218

QQT

QPA

QNQQT

QNLQT

GHESPHKSGQN
3094
KHDSPHKSGQN
3135
GHDSPHKSG
3177
GHDSPHKMA
3219

QQS

QQT

QSQPT

HNQQT

GHDSPHKSLQN
3095
GHDSPHKSGQT
3136
GHESPHKSG
3178
GHDSPHKHG
3220

QQT

QQT

QNEQT

QNQQN

GHDSPHKSGRN
309€
GPDSPHKIGQNQ
3137
GHDSPHKSA
3179
GHDSPHKSV
3221

QET

QS

QNQQT

QSQQS

GHDSPHKSGQT
3097
GHDSPHKSVQN
3138
GHDSPHKSG
3180
GHDSPHKSG
3222

QLT

QQT

QNQLT

QTVCT

GHDSPHKSEKN
3098
GHDSPHKSRQD
3139
GHDSPHKSA
3181
GQDSPHKSG
3223

QQT

QHT

QNLLT

QYQQI

GRDSPHKSGQD
3099
GPDSPHKSGQK
3140
THDSPHKSG
3182
GHDSPHKSG
3224

QQT

QQT

QNQQT

QQIMT

GHDSPHKTGHN
3100
GHDSPHKSRQS
3141
GNGSPHKRG
3183
GHDSPHKSR
3225

QQT

QHT

QNQQT

QNEQS

AHDSPHKSGQN
3101
GHDSPHKSVQN
3142
GHDSPHKSG
3184
GHDSPHKSG
3226

QLT

QLT

QSLQT

LNHQT

GYDSPHKSGQT
3102
AHDSPHKSGQN
3143
GHDSPHKSA
3185
GYDSPHKSG
3227

QQT

QQT

QNHQT

QNQQT

GHESPHKSGQT
3103
GQDSPHKSGQN
3144
GHDSPHKSG
3186
GHDSPHKSG
3228

QQI

QQS

RNRQT

QNLQT

GHDSPHKSGQS
3104
GHDSPHKSGRN
3145
EHDSPHKSG
3187
GHDSPHKSR
3229

KQA

QQI

QNQQT

QDQQT

GHESPHKSGQN
3105
GYDSPHKSGQY
3146
GHDSPHKYG
3188
GDDSPHKSG
3230

QQT

QHT

QNEQT

QKQLT

GHDSPHKTGQN
3106
GHDSPHKSRQN
3147
RHDSPHKSG
3189
GDDSPHKSG
3231

QPP

QQT

HNQQT

HNQQT

GHDSPHKSGQSP
3107
GHDSPHKSWVR
3148
GHDSPHKSQ
3190
GHDSPHKSG
3232

QT

QQT

QNQQT

QMIHT

GNDSPHKSVQN
3108
GHESPHKSGQN
3149
GHDSPHKSG
3191
GHDSPHKSG
3233

QQT

QHS

QNQQI

RNHQS

GHDSPHKSAQN
3109
GHDSPHKIGHN
3150
QHDSPHKSG
3192
GHDSPHKSV
3234

YQT

QQT

QNQQT

QNRQT

AHDSPHKIGQN
3110
WHDSPHKSGQN
3151
RHDSPHKIVQ
3193
GHDSPHKSG
3235

HQT

QQT

NQQT

QKMNT

GHESPHKSAQN
3111
RHDSPHKSGQN
3152
YHDSPHKSG
3194
GHDSPHKSG
3236

RQT

QQT

QNQQT

QSQQN

GHDSPHKSGQN
3112
GHDSPHKSNAW
3153
GHDSPHKSV
3195
GHDSPHKSG
3237

QQG

QQT

QSKQT

QYQHA

GQDSPHKIGQN
3113
GHDSPHKSGQS
3154
GNDSPHKIG
3196
GHDSPHKSG
3117

QQT

VPT

HNQQT

QNQWT

GHDSPHKSGQN
3114
GHESPHKSGQNI
3155
HHDSPHKSG
3197
GHDSPHKRG
3158

HLT

QP

QNQQT

PDQQS

GHDSPHKSGQY
3115
GHDSPHKSVQN
3156
GHDSPHKSG
3198
GHDSPHKSG
3200

QHT

HLN

QTQQI

RDQKT

GNDSPHKSVQN
3116
GHDSPHKIGLDQ
3157
GQDSPHKSG
3199
GHDSPHKSV
3201

HQT

QT

QNPLT

HNQQN

GHDSPHKSVQN
3118
GHDSPHKSGES
3159
KTISKRGSPH
4098
KGLGGSGSP
4099

QHT

QQT

SKAQNQQT

HSKAQNQQT

KTINGHDSPHSK
4100
KTINGSGSPHSK
4196
KEIYGSGSPH
4292
KTINGSGSPH
4388

AQNLQT

TCIQQT

SKAQNQQT

KRGQKQQT

KTINGHDSPHSK
4101
KTINGSGSPHSK
4197
KELSGSGSPH
4293
KTINGSGSPH
4389

AQNQQI

WLTQQT

SKAQNQQT

KRGQNQET

KTINGSGSPHFT
4102
KTINGSGSPHSK
4198
KETIGSGSPH
4294
KTINGSGSPH
4390

RQNQQT

WVVQQT

SKAQNQQT

KRGQNQLT

KTINGSGSPHSL
4103
KTINGSGSPHSK
4199
KEVLGSGSP
4295
KTINGSGSPH
4391

PWNQQT

YRLQQT

HSKAQNQQT

KRGRNQQT

KTINGHDSPHSK
4104
KTINGSGSPHSK
4200
KFALGHDSP
4296
KTINGSGSPH
4392

AQNHQT

YSKQQT

HKSGQKQQT

KSGGNQQT

KTMNGHDSPHS
4105
KTINGSGSPHSK
4201
KIINGHDSPH
4297
KTINGSGSPH
4393

KAQNQQT

YSRQQT

KSGQNLVL

KSGHNQET

KPYKGSGSPHSK
4106
KTINGSGSPHSL
4202
KIINGHDSPH
4298
KTINGSGSPH
4394

AQNQQT

KRNQQT

KSGQRNYT

KSGHNQLT

KRLWGSGSPHS
4107
KTINGSGSPHSL
4203
KIINGHDSPH
4299
KTINGSGSPH
4395

KAQNQQT

WFNQQT

SKAQNQQT

KSGHNQQN

KRMRGSGSPHS
4108
KTINGSGSPHSL
4204
KLNPGHDSP
4300
KTINGSGSPH
4396

KAQNQQT

WPNQQT

HKSGQTQQT

KSGLNQLT

KRTYGSGSPHSK
4109
KTINGSGSPHSL
4205
KLNRGHDSP
4301
KTINGSGSPH
4397

AQNQQT

WTNQQT

HKSGQNQQS

KSGPNQQT

KTINCLRSPHSK
4110
KTINGSGSPHSM
4206
KLSSGHDSPH
4302
KTINGSGSPH
4398

AQNQQT

RRNQQT

KSGQNQQN

KSGQGQQT

KTINFSRSPHSK
4111
KTINGSGSPHSP
4207
KNINGHDSP
4303
KTINGSGSPH
4399

AQNQQT

CLNQQT

HSKAQNQQT

KSGQHLQT

KTINGLRSPHFK
4112
KTINGSGSPHSQ
4208
KNNDGSGSP
4304
KTINGSGSPH
4400

AQNQQT

WQNQQT

HSKAQNQQT

KSGQHQQT

KTINGNRSPHNK
4113
KTINGSGSPHSR
4209
KNVMGSGSP
4305
KTINGSGSPH
4401

AQNQQT

CANQQT

HSKAQNQQT

KSGQKHQT

KTINGPRSPHYK
4114
KTINGSGSPHSRI
4210
KPINGHDSPH
4306
KTINGSGSPH
4402

AQNQQT

RNQQT

KSGQNKLS

KSGQKQQS

KTINGQASPHW
4115
KTINGSGSPHSR
4211
KPINGHDSPH
4307
KTINGSGSPH
4403

KAQNQQT

KSNQQT

KSGQNLSS

KSGQNEQT

KTINGRCSPHSK
4116
KTINGSGSPHSR
4212
KPINGHDSPH
4308
KTINGSGSPH
4404

AQNQQT

LWNQQT

SKAQNQQT

KSGQNHQT

KTINGRHSPHSK
4117
KTINGSGSPHSR
4213
KRINGHDSPH
4309
KTINGSGSPH
4405

AQNQQT

RFNQQT

SKAQNQQT

KSGQNKQT

KTINGRKSPHRK
4118
KTINGSGSPHSR
4214
KSCSGHDSP
4310
KTINGSGSPH
4406

AQNQQT

RPNQQT

HKSGQNQQS

KSGQNKTS

KTINGRLSPHW
4119
KTINGSGSPHSR
4215
KSINGHDSPH
4311
KTINGSGSPH
4407

KAQNQQT

SCNQQT

KSGQNLAS

KSGQNQEA

KTINGRLSPHYK
4120
KTINGSGSPHSR
4216
KSINGHDSPH
4312
KTINGSGSPH
4408

AQNQQT

SKNQQT

KSGQNLFL

KSGQNQET

KTINGRPSPHMK
4121
KTINGSGSPHSR
4217
KSINGHDSPH
4313
KTINGSGSPH
4409

AQNQQT

TKNQQT

KSGQNLLM

KSGQNQKT

KTINGRSSPHWK
4122
KTINGSGSPHSR
4218
KSINGHDSPH
4314
KTINGSGSPH
4410

AQNQQT

WLNQQT

KSGQNLLQ

KSGQNQQI

KTINGRWSPHSK
4123
KTINGSGSPHSS
4219
KSINGHDSPH
4315
KTINGSGSPH
4411

AQNQQT

VCNQQT

KSGQNSLG

KSGQNQQR

KTINGSGSPHAP
4124
KTINGSGSPHSS
4220
KSINGHDSPH
4316
KTINGSGSPH
4412

CQNQQT

WRNQQT

KSGQNTLQ

KSGQNQRT

KTINGSGSPHA
4125
KTINGSGSPHSV
4221
KSINGHDSPH
4317
KTINGSGSPH
4413

WAQNQQT

CQNQQT

KSSSNQQT

KSGQNQYT

KTINGSGSPHCM
4126
KTINGSGSPHSV
4222
KSINGHDSPH
4318
KTINGSGSPH
4414

RQNQQT

LCNQQT

KYKLNQQT

KSGQRQQT

KTINGSGSPHFC
4127
KTINGSGSPHSV
4223
KSINGSGSPH
4319
KTINGSGSPH
4415

SQNQQT

RRNQQT

KSGQKQQT

KSGQSQQT

KTINGSGSPHFL
4128
KTINGSGSPHSV
4224
KSINGSGSPH
4320
KTINGSGSPH
4416

FQNQQT

SCNQQT

KSGQNQQT

KSGQYQRT

KTINGSGSPHFW
4129
KTINGSGSPHSW
4225
KSINGSGSPH
4321
KTINGSGSPH
4417

AQNQQT

ALNQQT

SKAQGLST

KSGRNQQA

KTINGSGSPHLC
4130
KTINGSGSPHSW
4226
KSINGSGSPH
4322
KTINGSGSPH
4418

AQNQQT

ITNQQT

SKAQLLGT

KSRHNQQT

KTINGSGSPHLR
4131
KTINGSGSPHSW
4227
KSINGSGSPH
4323
KTINGSGSPH
4419

YQNQQT

PMNQQT

SKTSWQQT

KSRQYQQT

KTINGSGSPHLY
4132
KTINGSGSPHSW
4228
KSMNGHDSP
4324
KTINGSGSPH
4420

YQNQQT

RSNQQT

HSKAQNQQT

RKAQAPGT

KTINGSGSPHPL
4133
KTINGSGSPHSY
4229
KSTLGSGSPH
4325
KTINGSGSPH
4421

CQNQQT

FLNQQT

SKAQNQHT

SKAAMKQT

KTINGSGSPHRI
4134
KTINGSGSPHSY
4230
KSTLGSGSPH
4326
KTINGSGSPH
4422

RQNQQT

TYNQQT

SKAQNQQN

SKAGRQQT

KTINGSGSPHRL
4135
KTINGSGSPHSY
4231
KSTVGSGSPH
4327
KTINGSGSPH
4423

FQNQQT

WQNQQT

SKAQTQQT

SKAGRTQT

KTINGSGSPHSC
4136
KTINGSGSPHTL
4232
KTCKESGSPH
4328
KTINGSGSPH
4424

GQNQQT

CQNQQT

SKAQNQQT

SKAKSNQT

KTINGSGSPHSC
4137
KTINGSGSPHWL
4233
KTCKGSGSP
4329
KTINGSGSPH
4425

LRNQQT

RQNQQT

HSKAQNQQT

SKALKTQT

KTINGSGSPHSC
4138
KTINGSGSPHWP
4234
KTCKSSGSPH
4330
KTINGSGSPH
4426

LSNQQT

SQNQQT

SKAQNQQT

SKAPRTQT

KTINGSGSPHSC
4139
KTINGSGSPHYL
4235
KTDMGSGSP
4331
KTINGSGSPH
4427

RLNQQT

RQNQQT

HSKAQNQQT

SKAQAART

KTINGSGSPHSC
4140
KTINGSGSPHYT
4236
KTDNGIGSPH
4332
KTINGSGSPH
4428

SLNQQT

RQNQQT

SKAQNQQT

SKAQAILT

KTINGSGSPHSK
4141
KTINGSLSPHLW
4237
KTEGGSGSP
4333
KTINGSGSPH
4429

ACTLQT

AQNQQT

HSKAQNQQT

SKAQCRGT

KTINGSGSPHSK
4142
KTINGSPSPHCQ
4238
KTEHHSGSP
4334
KTINGSGSPH
4430

AFRAQT

AQNQQT

HSKAQNQQT

SKAQGLRT

KTINGSGSPHSK
4143
KTINGSRSPHLC
4239
KTEKDSGSP
4335
KTINGSGSPH
4431

AIRKQT

AQNQQT

HSKAQNQQT

SKAQKGVL

KTINGSGSPHSK
4144
KTINGSRSPHWR
4240
KTELGHDSP
4336
KTINGSGSPH
4432

AQASRT

AQNQQT

HKRGQNQQT

SKAQKSNT

KTINGSGSPHSK
4145
KTINGSVSPHWL
4241
KTESVSGSPH
4337
KTINGSGSPH
4433

AQFELT

AQNQQT

SKAQNQQT

SKAQNNKF

KTINGSGSPHSK
4146
KTINGTFSPHRK
4242
KTETNSGSPH
4338
KTINGSGSPH
4434

AQIVIT

AQNQQT

SKAQNQQT

SKAQNRRT

KTINGSGSPHSK
4147
KTINGWTSPHR
4243
KTETYSGSPH
4339
KTINGSGSPH
4435

AQLART

KAQNQQT

SKAQNQQT

SKAQPKQT

KTINGSGSPHSK
4148
KTINRGISPHSK
4244
KTEWLSGSP
4340
KTINGSGSPH
4436

AQLQRT

AQNQQT

HSKAQNQQT

SKAQRAPT

KTINGSGSPHSK
4149
KTINTVRSPHSK
4245
KTFNGSGSPH
4341
KTINGSGSPH
4437

AQNARR

AQNQQT

KSGQNQQT

SKAQREHT

KTINGSGSPHSK
4150
KTKLRSGSPHSK
4246
KTGLRHDSP
4342
KTINGSGSPH
4438

AQNCPR

AQNQQT

HKSGQKQQT

SKAQRFGT

KTINGSGSPHSK
4151
KTRLRSGSPHSK
4247
KTGLRHDSP
4343
KTINGSGSPH
4439

AQNMRR

AQNQQT

HKSGQNQQS

SKAQRPCT

KTINGSGSPHSK
4152
KWLLGSGSPHS
4248
KTGVTHDSP
4344
KTINGSGSPH
4440

AQNRRV

KAQNQQT

HKSGQKQQT

SKAQRQAT

KTINGSGSPHSK
4153
KWSQGSGSPHS
4249
KTIDGHESPH
4345
KTINGSGSPH
4441

AQPSRT

KAQNQQT

SKAQNQQT

SKAQRQPT

KTINGSGSPHSK
4154
KWYLGSGSPHS
4250
KTIEGHDSPH
4346
KTINGSGSPH
4442

AQQVKT

KAQNQQT

KSGQTQQT

SKAQTKLT

KTINGSGSPHSK
4155
KYHSGSGSPHS
4251
KTIHGHDSPH
4347
KTINGSGSPH
4443

AQQVRT

KAQNQQT

SKAQNQQT

SKAQTTHT

KTINGSGSPHSK
4156
KYLPGSGSPHSK
4252
KTIHGHESPH
4348
KTINGSGSPH
4444

AQRLKT

AQNQQT

SKAQNQQT

SKAQVQRT

KTINGSGSPHSK
4157
KAINGGGSPHSK
4253
KTIIGHDSPH
4349
KTINGSGSPH
4445

AQRRAT

TQNQQT

KSGQNRSS

SKAQVVRT

KTINGSGSPHSK
4158
KAINGHDSPHK
4254
KTIIGHDSPH
4350
KTINGSGSPH
4446

AQRRGT

RSPNQQT

KSGQRLGT

SKAQWPNT

KTINGSGSPHSK
4159
KAINGHDSPHKS
4255
KTIIGSGSPH
4351
KTINGSGSPH
4447

AQRRRT

FSPQQT

KSGQNQQT

SKAQYPST

KTINGSGSPHSK
4160
KAINGHDSPHKS
4256
KTIKGHDSPH
4352
KTINGSGSPH
4448

AQRTRT

GENQQP

KSGQNMLF

SKARALQT

KTINGSGSPHSK
4161
KAINGHDSPHKS
4257
KTILGSGSPH
4353
KTINGSGSPH
4449

AQRVHT

GQLART

SKAQNLQT

SKARDQHT

KTINGSGSPHSK
4162
KAINGHDSPHKS
4258
KTINGCSSPH
4354
KTINGSGSPH
4450

AQTYRT

GQNAFL

WKAQNQQT

SKARFQQT

KTINGSGSPHSK
4163
KAINGHDSPHKS
4259
KTINGGGSTH
4355
KTINGSGSPH
4451

AQVRKT

GQNAYT

SKAQNQQT

SKARRTQT

KTINGSGSPHSK
4164
KAINGHDSPHKS
4260
KTINGHDSPH
4356
KTINGSGSPH
4452

ARGRQT

GQNFAS

KAGQSQQT

SKARSLQT

KTINGSGSPHSK
4165
KAINGHDSPHKS
4261
KTINGHDSPH
4357
KTINGSGSPH
4453

ARLCQT

GQNLAS

KRGQNVPS

SKARVIQT

KTINGSGSPHSK
4166
KAINGHDSPHKS
4262
KTINGHDSPH
4358
KTINGSGSPH
4454

ARLKQT

GQNLGS

KRGRSYQT

SKAWYLQT

KTINGSGSPHSK
4167
KAINGHDSPHKS
4263
KTINGHDSPH
4359
KTINGSGSPH
1455

ARNSQT

GQNLKF

KTGQNPPT

SKGGGQQT

KTINGSGSPHSK
4168
KAINGHDSPHKS
4264
KTINGHDSPH
4360
KTINGSGSPH
4456

ARWVQT

GQNLLK

SKAENQQT

SKGSRQQT

KTINGSGSPHSK
4169
KAINGHDSPHKS
4265
KTINGHDSPH
4361
KTINGSGSPH
4457

AVRWQT

GQNLSR

SKALSLQT

SKLQRQQT

KTINGSGSPHSK
4170
KAINGHDSPHKS
4266
KTINGHDSPH
4362
KTINGSGSPH
4458

AYTRQT

GQNLSS

SKAQGQQT

SKMLRQQT

KTINGSGSPHSK
4171
KAINGHDSPHKS
4267
KTINGHDSPH
4363
KTINGSGSPH
4459

CQSQQT

GQNSLG

SKAQHQQT

SKSSIKQT

KTINGSGSPHSK
4172
KAINGHDSPHKS
4268
KTINGHDSPH
4364
KTINGSGSPH
4460

FLRQQT

GQNTLQ

SKAQIQQT

SKVRFQQT

KTINGSGSPHSK
4173
KAINGHDSPHKS
4269
KTINGHDSPH
4365
KTINGSGSPH
4461

FRFQQT

GQNTSL

SKAQKQQT

SVVWNQQT

KTINGSGSPHSK
4174
KAINGHDSPHKS
4270
KTINGHDSPH
4366
KTINGSTSPH
4462

FRLQQT

GQRLGT

SKAQNLSS

KLAQNQQP

KTINGSGSPHSK
4175
KAINGHDSPHKS
4271
KTINGHDSPH
4367
KTINRHDSPH
4463

FRRQQT

GQRNYT

SKAQNPQT

KSGQRPST

KTINGSGSPHSK
4176
KAINGHDSPHKS
4272
KTINGHDSPH
4368
KTINRIMSPH
4464

GMKQQT

GQRPST

SKAQNQET

SKAQNQQT

KTINGSGSPHSK
4177
KAINGHDSPHKS
4273
KTINGHDSPH
4369
KTINTARSPH
4465

KLRQQT

GQRPVT

SKAQNQHT

SKAQNQQT

KTINGSGSPHSK
4178
KAINGHDSPHKS
4274
KTINGHDSPH
4370
KTISGHDSPH
4466

KRPQQT

GQVPST

SKAQNQLT

SKAQNQQT

KTINGSGSPHSK
4179
KAINGHDSPHKS
4275
KTINGHDSPH
4371
KTISGSGSPH
4467

KSRQQT

LSNQQT

SKAQNQPT

KSGQNQQT

KTINGSGSPHSK
4180
KAINGHDSPHKS
4276
KTINGHDSPH
4372
KTITGHDSPH
4468

LYRQQT

VLSQQT

SKAQNQQA

KSGQRLGT

KTINGSGSPHSK
4181
KAINGHDSPHKT
4277
KTINGHDSPH
4373
KTITGSGSPH
4469

LYWQQT

LQNQQT

SKAQNTGS

KSGQNQQT

KTINGSGSPHSK
4182
KAINGHNSPHSK
4278
KTINGHDSPH
4374
KTIYGHDSP
4470

PRMQQT

AQNQQT

SKAQSQQT

HKSGQRLGT

KTINGSGSPHSK
4183
KAINGLDSPHSK
4279
KTINGHDSPH
4375
KTLNGHDSP
4471

RFPQQT

AQNQQT

SKAQTQQT

HKSGQNLFL

KTINGSGSPHSK
4184
KAINGSGSPHKS
4280
KTINGHDSPH
4376
KTLNGHDSP
4472

RFRQQT

GQNQQT

SKAQYQQT

HKSGQNLSS

KTINGSGSPHSK
4185
KAINGSGSPHSK
4281
KTINGHDSPH
4377
KTLSFHDSPH
4473

RPYQQT

AQGQQT

SKARNQQT

KSGQNQQS

KTINGSGSPHSK
4186
KAINGSGSPHSK
4282
KTINGHDSPH
4378
KTSNGSGSP
4474

RRMQQT

AQLSGT

SKLPGQQT

HSKAQNTMT

KTINGSGSPHSK
4187
KAINGSGSPHSK
4283
KTINGHDSPH
4379
KTTNGHDSP
4475

RSKQQT

AQNGSL

SKSPNQQT

HSKAQNQQT

KTINGSGSPHSK
4188
KAINGSGSPHSK
4284
KTINGHESPH
4380
KTVNGGGSP
4476

RSRQQT

AQNSLL

KSGQNAFL

HSKAQNQQT

KTINGSGSPHSK
4189
KAINGSGSPHSK
4285
KTINGIGSPH
4381
KTVNGHDSP
4477

RTMQQT

AVGLQT

SKAPNEQT

HKSGQNVSL

KTINGSGSPHSK
4190
KAINGSGSPHSK
4286
KTINGQDSPH
4382
KTVNGHDSP
4478

RTRQQT

SLLQQT

KSGQNLHM

HKSGQRPST

KTINGSGSPHSK
4191
KAINGSGSPHSK
4287
KTINGRGSPH
4383
KTVNGHDSP
4479

RVRQQT

SLPQQT

SKAQIGMT

HKSGQTQQA

KTINGSGSPHSK
4192
KAINGSGSPHSK
4288
KTINGRGSPH
4384
KTVNGHESP
4480

RYIQQT

STFQQT

SKAQNQVL

HSKAQNQQT

KTINGSGSPHSK
4193
KAITGHDSPHSK
4289
KTINGRGSPH
4385
KTVNGSGSP
4481

RYNQQT

AQNQQT

SKAQSPTT

HSKAQGLST

KTINGSGSPHSK
4194
KDVMGSGSPHS
4290
KTINGRGSPH
4386
KTVNGSGSP
4482

RYPQQT

KAQNQQT

SKATSFQT

HSKAQNVTS

KTINGSGSPHSK
4195
KEIVGSGSPHSK
4291
KTINGSGSPH
4387
KTVPASGSP
4483

RYSQQT

AQNQQT

FVVQNQQT

HSKAQNQQT

NTINGSGSPHSK
4484
TTINGGGSPHSK
4485

AHNQQT

AQNQQT

TABLE 2A

Exemplary Peptide Sequences

SEQ

SEQ

ID
Amino Acid
ID

NO:
Sequence
NO:
Nucleotide Sequence

941
SPHSKA
942
AGCCCACACAGCAAAGCA

943
HDSPHKSG
944
CACGACAGCCCACACAAAAG

CGGA

2
HDSPHK
3
CACGACAGCCCACACAAA

TABLE 2B

Exemplary Peptide Sequences

Amino
SEQ
Amino
SEQ
Amino
SEQ
Amino
SEQ

Acid
ID
Acid
ID
Acid
ID
Acid
ID

Sequence
NO:
Sequence
NO:
Sequence
NO:
Sequence
NO:

SPHSKA
945
SPHKKN
954
SPHKTS
963
SPHTRG
972

SPHKSG
946
SPHVRM
955
SPHKTT
964
SPHVRG
973

SPHARM
947
SPHRKA
956
SPHKTY
965
SPHKRG
974

SPHVKS
948
SPHKFG
957
SPHKYG
966
SPHGAR
975

SPHASR
949
SPHKIG
958
SPHSKD
967
SPHRSG
976

SPHVKI
950
SPHKLG
959
SPHSKP
968
SPHKSA
977

SPHKSR
951
SPHSKL
960
SPHSRA
969
SPHSKR
978

SPHSLR
952
SPHSRG
961
SPHSSR
970
SPHFLR
979

SPHSKW
953
SPHSKS
962
SPHWKA
971
SPHVRR
980

STHASR
985
SQHKSG
986
HDSPHK
2
HDSPHSKA
4486

In some embodiments, the peptide comprises an amino acid sequence having the formula [N1]-[N2]-[N3], wherein [N42] comprises the amino acid sequence of SPH and [N43] comprises X4, X5, and X6, wherein at least one of X4, X5, or X6 is a basic amino acid, e.g., a K or R. In some embodiments, position X4 of [N42] is K. In some embodiments, position X5 of [N42] is K.

In some embodiments, [N1] comprises X1, X2, and X3, wherein at least one of X1, X2, or X3 is G. In some embodiments, position X1 of [N1] is G, V, R, D, E, M, T, I, S, A, N, L, K, H, P, W, or C. In some embodiments, position X2 of [N41] is: S, V, L, N, D, H, R, P, G, T, I, A, E, Y, M, or Q. In some embodiments, position X3 of [N41] is: G, C, L, D, E, Y, H, V, A, N, P, or S. In some embodiments, [N1] comprises GS, SG, GH, HD, GQ, QD, VS, CS, GR, RG, QS, SH, MS, RN, TS, IS, GP, ES, SS, GN, AS, NS, LS, GG, KS, GT, PS, RS, GI, WS, DS, ID, GL, DA, DG, ME, EN, KN, KE, AI, NG, PG, TG, SV, IG, LG, AG, EG, SA, YD, HE, HG, RD, ND, PD, MG, QV, DD, HN, HP, GY, GM, GD, or HS. In some embodiments, [N1] comprises GS, SG, GH, or HD. In some embodiments [N1] is or comprises GSG, GHD, GQD, VSG, CSG, GRG, CSH, GQS, GSH, RVG, GSC, GLL, GDD, GHE, GNY, MSG, RNG, TSG, ISG, GPG, ESG, SSG, GNG, ASG, NSG, LSG, GGG, KSG, HSG, GTG, PSG, GSV, RSG, GIG, WSG, DSG, IDG, GLG, DAG, DGG, MEG, ENG, GSA, KNG, KEG, AIG, GYD, GHG, GRD, GND, GPD, GMG, GQV, GHN, GHP, or GHS. In some embodiments, [N1] is or comprises GSG. In some embodiments, [N1] is or comprises GHD. In some embodiments, [N1]-[N2] comprises SGSPH (SEQ ID NO: 4752), HDSPH (SEQ ID NO: 4703), QDSPH (SEQ ID NO: 4753), RGSPH (SEQ ID NO: 4754), SHSPH (SEQ ID NO: 4755), QSSPH (SEQ ID NO: 4756), DDSPH (SEQ ID NO: 4757), HESPH (SEQ ID NO: 4758), NYSPH (SEQ ID NO: 4759), VGSPH (SEQ ID NO: 4760), SCSPH (SEQ ID NO: 4761), LLSPH (SEQ ID NO: 4762), NGSPH (SEQ ID NO: 4763), PGSPH (SEQ ID NO: 4764), GGSPH (SEQ ID NO: 4765), TGSPH (SEQ ID NO: 4766), SVSPH (SEQ ID NO: 4767), IGSPH (SEQ ID NO: 4768), DGSPH (SEQ ID NO: 4769), LGSPH (SEQ ID NO: 4770), AGSPH (SEQ ID NO: 4771), EGSPH (SEQ ID NO: 4772), SASPH (SEQ ID NO: 4773), YDSPH (SEQ ID NO: 4774), HGSPH (SEQ ID NO: 4775), RDSPH (SEQ ID NO: 4776), NDSPH (SEQ ID NO: 4777), PDSPH (SEQ ID NO: 4778), MGSPH (SEQ ID NO: 4779), QVSPH (SEQ ID NO: 4780), HNSPH (SEQ ID NO: 4781), HPSPH (SEQ ID NO: 4782), or HSSPH (SEQ ID NO: 4783); an amino acid sequence comprising any portion of any of the aforesaid amino acid sequences (e.g., any 2, 3, or 4 amino acids, e.g., consecutive amino acids) thereof; an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the aforesaid amino acid sequences; or an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the aforesaid amino acid sequences. In some embodiments, [N1]-[N2] is or comprises GSGSPH (SEQ ID NO: 4695), GHDSPH (SEQ ID NO: 4784), GQDSPH (SEQ ID NO: 4785), VSGSPH (SEQ ID NO: 4786), CSGSPH (SEQ ID NO: 4787), GRGSPH (SEQ ID NO: 4788), CSHSPH (SEQ ID NO: 4789), GQSSPH (SEQ ID NO: 4790), GSHSPH (SEQ ID NO: 4791), GDDSPH (SEQ ID NO: 4792), GHESPH (SEQ ID NO: 4793), GNYSPH (SEQ ID NO: 4794), RVGSPH (SEQ ID NO: 4795), GSCSPH (SEQ ID NO: 4796), GLLSPH (SEQ ID NO: 4797), MSGSPH (SEQ ID NO: 4798), RNGSPH (SEQ ID NO: 4799), TSGSPH (SEQ ID NO: 4800), ISGSPH (SEQ ID NO: 4801), GPGSPH (SEQ ID NO: 4802), ESGSPH (SEQ ID NO: 4803), SSGSPH (SEQ ID NO: 4804), GNGSPH (SEQ ID NO: 4805), ASGSPH (SEQ ID NO: 4806), NSGSPH (SEQ ID NO: 4807), LSGSPH (SEQ ID NO: 4808), GGGSPH (SEQ ID NO: 4809), KSGSPH (SEQ ID NO: 4810), HSGSPH (SEQ ID NO: 4811), GTGSPH (SEQ ID NO: 4812), PSGSPH (SEQ ID NO: 4813), GSVSPH (SEQ ID NO: 4814), RSGSPH (SEQ ID NO: 4815), GIGSPH (SEQ ID NO: 4816), WSGSPH (SEQ ID NO: 4817), DSGSPH (SEQ ID NO: 4818), IDGSPH (SEQ ID NO: 4819), GLGSPH (SEQ ID NO: 4820), DAGSPH (SEQ ID NO: 4821), DGGSPH (SEQ ID NO: 4822), MEGSPH (SEQ ID NO: 4823), ENGSPH (SEQ ID NO: 4824), GSASPH (SEQ ID NO: 4825), KNGSPH (SEQ ID NO: 4826), KEGSPH (SEQ ID NO: 4827), AIGSPH (SEQ ID NO: 4828), GYDSPH (SEQ ID NO: 4829), GHGSPH (SEQ ID NO: 4830), GRDSPH (SEQ ID NO: 4831), GNDSPH (SEQ ID NO: 4832), GPDSPH (SEQ ID NO: 4833), GMGSPH (SEQ ID NO: 4834), GQVSPH (SEQ ID NO: 4835), GHNSPH (SEQ ID NO: 4836), GHPSPH (SEQ ID NO: 4837), or GHSSPH (SEQ ID NO: 4838); an amino acid sequence comprising any portion of any of the aforesaid amino acid sequences (e.g., any 2, 3, 4, or 5 amino acids, e.g., consecutive amino acids) thereof; an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the aforesaid amino acid sequences; or an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the aforesaid amino acid sequences. In some embodiments, [N1]-[N2] is or comprises GSGSPH (SEQ ID NO: 4695). In some embodiments, [N1]-[N2] is or comprises GHDSPH (SEQ ID NO: 4784).

In some embodiments, X4, X5, or both of [N3] are K. In some embodiments, X4, X5, or X6 of [N3] is R. In some embodiments, position X4 of [N3] is: A, K, V, S, T, G, F, W, V, N, or R. In some embodiments, position X5 of [N3] is: S, K, T, F, I, L, Y, H, M, or R. In some embodiments, position X6 of [N3] is: G, R, A, M, I, N, T, Y, D, P, V, L, E, W, N, Q, K, or S. In some embodiments, [N3] comprises SK, KA, KS, AR, RM, VK, AS, SR, VK, KR, KK, KN, VR, RS, RK, KT, TS, KF, FG, KI, IG, KL, LG, TT, TY, KY, YG, KD, KP, TR, RG, VR, GA, SL, SS, FL, WK, SA, RA, LR, KW, RR, GK, TK, NK, AK, KV, KG, KH, KM, TG, SE, SV, SW, SN, HG, SQ, LW, MG, MA, or SG. In some embodiments, [N3] comprises SK, KA, KS, or SG. In some embodiments, [N3] is or comprises SKA, KSG, ARM, VKS, ASR, VKI, KKN, VRM, RKA, KTS, KFG, KIG, KLG, KTT, KTY, KYG, SKD, SKP, TRG, VRG, KRG, GAR, KSA, KSR, SKL, SRA, SKR, SLR, SRG, SSR, FLR, SKW, SKS, WKA, VRR, SKV, SKT, SKG, GKA, TKA, NKA, SKL, SKN, AKA, KTG, KSL, KSE, KSV, KSW, KSN, KHG, KSQ, KSK, KLW, WKG, KMG, KMA, or RSG. In some embodiments, [N3] is or comprises SKA. In some embodiments, [N3] is or comprises KSG. In some embodiments, [N2]-[N3] comprises SPHSK (SEQ ID NO: 4701), SPHKS (SEQ ID NO: 4704), SPHAR (SEQ ID NO: 4705), SPHVK (SEQ ID NO: 4706), SPHAS (SEQ ID NO: 4707), SPHKK (SEQ ID NO: 4708), SPHVR (SEQ ID NO: 4709), SPHRK (SEQ ID NO: 4710), SPHKT (SEQ ID NO: 4711), SPHKF (SEQ ID NO: 4712), SPHKI (SEQ ID NO: 4713), SPHKL (SEQ ID NO: 4714), SPHKY (SEQ ID NO: 4715), SPHTR (SEQ ID NO: 4716), SPHKR (SEQ ID NO: 4717), SPHGA (SEQ ID NO: 4718), SPHSR (SEQ ID NO: 4719), SPHSL (SEQ ID NO: 4720), SPHSS (SEQ ID NO: 4721), SPHFL (SEQ ID NO: 4722), SPHWK (SEQ ID NO: 4723), SPHGK (SEQ ID NO: 4724), SPHTK (SEQ ID NO: 4725), SPHNK (SEQ ID NO: 4726), SPHAK (SEQ ID NO: 4727), SPHKH (SEQ ID NO: 4728), SPHKM (SEQ ID NO: 4729), or SPHRS (SEQ ID NO: 4730). In some embodiments [N2]-[N3] comprises SPHSK (SEQ ID NO: 4701) or SPHKS (SEQ ID NO: 4704). In some embodiments, [N2]-[N3] is or comprises SPHSKA (SEQ ID NO: 941), SPHKSG (SEQ ID NO: 946), SPHARM (SEQ ID NO: 947), SPHVKS (SEQ ID NO: 948), SPHASR (SEQ ID NO: 949), SPHVKI (SEQ ID NO: 950), SPHKKN (SEQ ID NO: 954), SPHVRM (SEQ ID NO: 955), SPHRKA (SEQ ID NO: 956), SPHKFG (SEQ ID NO: 957), SPHKIG (SEQ ID NO: 958), SPHKLG (SEQ ID NO: 959), SPHKTS (SEQ ID NO: 963), SPHKTT (SEQ ID NO: 964), SPHKTY (SEQ ID NO: 965), SPHKYG (SEQ ID NO: 966), SPHSKD (SEQ ID NO: 967), SPHSKP (SEQ ID NO: 968), SPHTRG (SEQ ID NO: 972), SPHVRG (SEQ ID NO: 973), SPHKRG (SEQ ID NO: 974), SPHGAR (SEQ ID NO: 975), SPHKSA (SEQ ID NO: 977), SPHKSR (SEQ ID NO: 951), SPHSKL (SEQ ID NO: 960), SPHSRA (SEQ ID NO: 969), SPHSKR (SEQ ID NO: 978), SPHSLR (SEQ ID NO: 952), SPHSRG (SEQ ID NO: 961), SPHSSR (SEQ ID NO: 970), SPHFLR (SEQ ID NO: 979), SPHSKW (SEQ ID NO: 953), SPHSKS (SEQ ID NO: 962), SPHWKA (SEQ ID NO: 971), SPHVRR (SEQ ID NO: 980), SPHSKT (SEQ ID NO: 4731), SPHSKG (SEQ ID NO: 4732), SPHGKA (SEQ ID NO: 4733), SPHNKA (SEQ ID NO: 4734), SPHSKN (SEQ ID NO: 4735), SPHAKA (SEQ ID NO: 4736), SPHSKV (SEQ ID NO: 4737), SPHKTG (SEQ ID NO: 4738), SPHTKA (SEQ ID NO: 4739), SPHKSL (SEQ ID NO: 4740), SPHKSE (SEQ ID NO: 4741), SPHKSV (SEQ ID NO: 4742), SPHKSW (SEQ ID NO: 4743), SPHKSN (SEQ ID NO: 4744), SPHKHG (SEQ ID NO: 4745), SPHKSQ (SEQ ID NO: 4746), SPHKSK (SEQ ID NO: 4747), SPHKLW (SEQ ID NO: 4748), SPHWKG (SEQ ID NO: 4749), SPHKMG (SEQ ID NO: 4750), SPHKMA (SEQ ID NO: 4751), or SPHRSG (SEQ ID NO: 976). In some embodiments, [N2]-[N3] is or comprises SPHSKA (SEQ ID NO: 941). In some embodiments, [N2]-[N3] is or comprises SPHKSG (SEQ ID NO: 946).

In some embodiments, [N1]-[N2]-[N3] comprises SGSPHSK (SEQ ID NO: 4839), HDSPHKS (SEQ ID NO: 4840), SGSPHAR (SEQ ID NO: 4841), SGSPHVK (SEQ ID NO: 4842), QDSPHKS (SEQ ID NO: 4843), SGSPHKK (SEQ ID NO: 4844), SGSPHVR (SEQ ID NO: 4845), SGSPHAS (SEQ ID NO: 4846), SGSPHRK (SEQ ID NO: 4847), SGSPHKT (SEQ ID NO: 4848), SHSPHKS (SEQ ID NO: 4849), QSSPHRS (SEQ ID NO: 4850), RGSPHAS (SEQ ID NO: 4851), RGSPHSK (SEQ ID NO: 4852), SGSPHKF (SEQ ID NO: 4853), SGSPHKI (SEQ ID NO: 4854), SGSPHKL (SEQ ID NO: 4855), SGSPHKY (SEQ ID NO: 4856), SGSPHTR (SEQ ID NO: 4857), SHSPHKR (SEQ ID NO: 4858), SGSPHGA (SEQ ID NO: 4859), HDSPHKR (SEQ ID NO: 4860), DDSPHKS (SEQ ID NO: 4861), HESPHKS (SEQ ID NO: 4862), NYSPHKI (SEQ ID NO: 4863), SGSPHSR (SEQ ID NO: 4864), SGSPHSL (SEQ ID NO: 4865), SGSPHSS (SEQ ID NO: 4866), VGSPHSK (SEQ ID NO: 4867), SCSPHRK (SEQ ID NO: 4868), SGSPHFL (SEQ ID NO: 4869), LLSPHWK (SEQ ID NO: 4870), NGSPHSK (SEQ ID NO: 4871), PGSPHSK (SEQ ID NO: 4872), GGSPHSK (SEQ ID NO: 4873), TGSPHSK (SEQ ID NO: 4874), SVSPHGK (SEQ ID NO: 4875), SGSPHTK (SEQ ID NO: 4876), IGSPHSK (SEQ ID NO: 4877), DGSPHSK (SEQ ID NO: 4878), SGSPHNK (SEQ ID NO: 4879), LGSPHSK (SEQ ID NO: 4880), AGSPHSK (SEQ ID NO: 4881), EGSPHSK (SEQ ID NO: 4882), SASPHSK (SEQ ID NO: 4883), SGSPHAK (SEQ ID NO: 4884), HDSPHKI (SEQ ID NO: 4885), YDSPHKS (SEQ ID NO: 4886), HDSPHKT (SEQ ID NO: 4887), RGSPHKR (SEQ ID NO: 4888), HGSPHSK (SEQ ID NO: 4889), RDSPHKS (SEQ ID NO: 4890), NDSPHKS (SEQ ID NO: 4891), QDSPHKI (SEQ ID NO: 4892), PDSPHKI (SEQ ID NO: 4893), PDSPHKS (SEQ ID NO: 4894), MGSPHSK (SEQ ID NO: 4895), HDSPHKH (SEQ ID NO: 4896), QVSPHKS (SEQ ID NO: 4897), HNSPHKS (SEQ ID NO: 4898), NGSPHKR (SEQ ID NO: 4899), HDSPHKY (SEQ ID NO: 4900), NDSPHKI (SEQ ID NO: 4901), HDSPHKL (SEQ ID NO: 4902), HPSPHWK (SEQ ID NO: 4903), HDSPHKM (SEQ ID NO: 4904), or HSSPHRS (SEQ ID NO: 4905). In some embodiments, [N1]-[N2]-[N3] is GSGSPHSKA (SEQ ID NO: 4697), GHDSPHKSG (SEQ ID NO: 4698), GSGSPHARM (SEQ ID NO: 4906), GSGSPHVKS (SEQ ID NO: 4907), GQDSPHKSG (SEQ ID NO: 4908), GSGSPHASR (SEQ ID NO: 4909), GSGSPHVKI (SEQ ID NO: 4910), GSGSPHKKN (SEQ ID NO: 4911), GSGSPHVRM (SEQ ID NO: 4912), VSGSPHSKA (SEQ ID NO: 4913), CSGSPHSKA (SEQ ID NO: 4914), GSGSPHRKA (SEQ ID NO: 4915), CSGSPHKTS (SEQ ID NO: 4916), CSHSPHKSG (SEQ ID NO: 4917), GQSSPHRSG (SEQ ID NO: 4918), GRGSPHASR (SEQ ID NO: 4919), GRGSPHSKA (SEQ ID NO: 4920), GSGSPHKFG (SEQ ID NO: 4921), GSGSPHKIG (SEQ ID NO: 4922), GSGSPHKLG (SEQ ID NO: 4923), GSGSPHKTS (SEQ ID NO: 4924), GSGSPHKTT (SEQ ID NO: 4925), GSGSPHKTY (SEQ ID NO: 4926), GSGSPHKYG (SEQ ID NO: 4927), GSGSPHSKD (SEQ ID NO: 4928), GSGSPHSKP (SEQ ID NO: 4929), GSGSPHTRG (SEQ ID NO: 4930), GSGSPHVRG (SEQ ID NO: 4931), GSHSPHKRG (SEQ ID NO: 4932), GSHSPHKSG (SEQ ID NO: 4933), VSGSPHASR (SEQ ID NO: 4934), VSGSPHGAR (SEQ ID NO: 4935), VSGSPHKFG (SEQ ID NO: 4936), GHDSPHKRG (SEQ ID NO: 4937), GDDSPHKSG (SEQ ID NO: 4938), GHESPHKSA (SEQ ID NO: 4939), GHDSPHKSA (SEQ ID NO: 4940), GNYSPHKIG (SEQ ID NO: 4941), GHDSPHKSR (SEQ ID NO: 4942), GSGSPHSKL (SEQ ID NO: 4943), GSGSPHSRA (SEQ ID NO: 4944), GSGSPHSKR (SEQ ID NO: 4945), GSGSPHSLR (SEQ ID NO: 4946), GSGSPHSRG (SEQ ID NO: 4947), GSGSPHSSR (SEQ ID NO: 4948), RVGSPHSKA (SEQ ID NO: 4949), GSCSPHRKA (SEQ ID NO: 4950), GSGSPHFLR (SEQ ID NO: 4951), GSGSPHSKW (SEQ ID NO: 4952), GSGSPHSKS (SEQ ID NO: 4953), GLLSPHWKA (SEQ ID NO: 4954), GSGSPHVRR (SEQ ID NO: 4955), GSGSPHSKV (SEQ ID NO: 4956), MSGSPHSKA (SEQ ID NO: 4957), RNGSPHSKA (SEQ ID NO: 4958), TSGSPHSKA (SEQ ID NO: 4959), ISGSPHSKA (SEQ ID NO: 4960), GPGSPHSKA (SEQ ID NO: 4961), GSGSPHSKT (SEQ ID NO: 4962), ESGSPHSKA (SEQ ID NO: 4963), SSGSPHSKA (SEQ ID NO: 4964), GNGSPHSKA (SEQ ID NO: 4965), ASGSPHSKA (SEQ ID NO: 4966), NSGSPHSKA (SEQ ID NO: 4967), LSGSPHSKA (SEQ ID NO: 4968), GGGSPHSKA (SEQ ID NO: 4969), KSGSPHSKA (SEQ ID NO: 4970), GGGSPHSKS (SEQ ID NO: 4971), GSGSPHSKG (SEQ ID NO: 4972), HSGSPHSKA (SEQ ID NO: 4973), GTGSPHSKA (SEQ ID NO: 4974), PSGSPHSKA (SEQ ID NO: 4975), GSVSPHGKA (SEQ ID NO: 4976), RSGSPHSKA (SEQ ID NO: 4977), GSGSPHTKA (SEQ ID NO: 4978), GIGSPHSKA (SEQ ID NO: 4979), WSGSPHSKA (SEQ ID NO: 4980), DSGSPHSKA (SEQ ID NO: 4981), IDGSPHSKA (SEQ ID NO: 4982), GSGSPHNKA (SEQ ID NO: 4983), GLGSPHSKS (SEQ ID NO: 4984), DAGSPHSKA (SEQ ID NO: 4985), DGGSPHSKA (SEQ ID NO: 4986), MEGSPHSKA (SEQ ID NO: 4987), ENGSPHSKA (SEQ ID NO: 4988), GSASPHSKA (SEQ ID NO: 4989), GNGSPHSKS (SEQ ID NO: 4990), KNGSPHSKA (SEQ ID NO: 4991), KEGSPHSKA (SEQ ID NO: 4992), AIGSPHSKA (SEQ ID NO: 4993), GSGSPHSKN (SEQ ID NO: 4994), GSGSPHAKA (SEQ ID NO: 4995), GHDSPHKIG (SEQ ID NO: 4996), GYDSPHKSG (SEQ ID NO: 4997), GHESPHKSG (SEQ ID NO: 4998), GHDSPHKTG (SEQ ID NO: 4999), GRGSPHKRG (SEQ ID NO: 5000), GQDSPHKSG (SEQ ID NO: 4908), GHDSPHKSL (SEQ ID NO: 5001), GHGSPHSKA (SEQ ID NO: 5002), GHDSPHKSE (SEQ ID NO: 5003), VSGSPHSKA (SEQ ID NO: 4913), GRDSPHKSG (SEQ ID NO: 5004), GNDSPHKSV (SEQ ID NO: 5005), GQDSPHKIG (SEQ ID NO: 5006), GHDSPHKSV (SEQ ID NO: 5007), GPDSPHKIG (SEQ ID NO: 5008), GPDSPHKSG (SEQ ID NO: 5009), GHDSPHKSW (SEQ ID NO: 5010), GHDSPHKSN (SEQ ID NO: 5011), GMGSPHSKT (SEQ ID NO: 5012), GHDSPHKHG (SEQ ID NO: 5013), GQVSPHKSG (SEQ ID NO: 5014), GDDSPHKSV (SEQ ID NO: 5015), GHNSPHKSG (SEQ ID NO: 5016), GNGSPHKRG (SEQ ID NO: 5017), GHDSPHKYG (SEQ ID NO: 5018), GHDSPHKSQ (SEQ ID NO: 5019), GNDSPHKIG (SEQ ID NO: 5020), GHDSPHKSK (SEQ ID NO: 5021), GHDSPHKLW (SEQ ID NO: 5022), GHPSPHWKG (SEQ ID NO: 5023), GHDSPHKMG (SEQ ID NO: 5024), GHDSPHKMA (SEQ ID NO: 5025), or GHSSPHRSG (SEQ ID NO: 5026); an amino acid sequence comprising any portion of any of the aforesaid amino acid sequences (e.g., any 2, 3, 4, 5, 6, 7, or 8 amino acids, e.g., consecutive amino acids) thereof; an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the aforesaid amino acid sequences; or an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the aforesaid amino acid sequences. In some embodiments, [N1]-[N2]-[N3] is or comprises GSGSPHSKA (SEQ ID NO: 4697). In some embodiments, [N1]-[N2]-[N3] is or comprises GHDSPHKSG (SEQ ID NO: 4698).

In some embodiments, the peptide comprising an amino acid sequence having the formula [N1]-[N2]-[N3], further comprises [N4] which comprises X7 X8 X9 X10. In some embodiments, position X7 of [N4] is W, Q, K, R, G, L, V, S, P, H, K, I, M, A, E, or F. In some embodiments, position X8 of [N4] is N, Y, C, K, T, H, R, D, V, S, P, G, W, E, F, A, I, M, Q, or L. In some embodiments, position X9 of [N4] is Q, G, K, H, R, T, L, D, A, P, I, F, V, M, W, Y, S, E, N, or Y. In some embodiments, position X10 of [N4] is Q, H, L, R, W, K, A, P, E, M, I, S, G, N, Y, C, V, T, D, or V. In some embodiments [N4] is or comprises QNQQ (SEQ ID NO: 5028), WNQQ (SEQ ID NO: 5029), QYYV (SEQ ID NO: 5030), RRQQ (SEQ ID NO: 5031), QNQQ (SEQ ID NO: 5028), GCGQ (SEQ ID NO: 5032), LRQQ (SEQ ID NO: 5033), RNQQ (SEQ ID NO: 5034), VNQQ (SEQ ID NO: 5035), FRLQ (SEQ ID NO: 5036), FNQQ (SEQ ID NO: 5037), LLQQ (SEQ ID NO: 5038), SNQQ (SEQ ID NO: 5039), RLQQ (SEQ ID NO: 5040), LNQQ (SEQ ID NO: 5041), QRKL (SEQ ID NO: 5042), LRRQ (SEQ ID NO: 5043), QRLR (SEQ ID NO: 5044), QRRL (SEQ ID NO: 5045), RRLQ (SEQ ID NO: 5046), RLRQ (SEQ ID NO: 5047), SKRQ (SEQ ID NO: 5048), QLYR (SEQ ID NO: 5049), QLTV (SEQ ID NO: 5050), QNKQ (SEQ ID NO: 5051), KNQQ (SEQ ID NO: 5052), QKQQ (SEQ ID NO: 5053), QTQQ (SEQ ID NO: 5054), QNHQ (SEQ ID NO: 5055), QHQQ (SEQ ID NO: 5056), QNQH (SEQ ID NO: 5057), QHRQ (SEQ ID NO: 5058), LTQQ (SEQ ID NO: 5059), QNQW (SEQ ID NO: 5060), QNTH (SEQ ID NO: 5061), RRRQ (SEQ ID NO: 5062), QYQQ (SEQ ID NO: 5063), QNDQ (SEQ ID NO: 5064), QNRH (SEQ ID NO: 5065), RDQQ (SEQ ID NO: 5066), PNLQ (SEQ ID NO: 5067), HVRQ (SEQ ID NO: 5068), PNQH (SEQ ID NO: 5069), HNQQ (SEQ ID NO: 5070), QSQQ (SEQ ID NO: 5071), QPAK (SEQ ID NO: 5072), QNLA (SEQ ID NO: 5073), QNQL (SEQ ID NO: 5074), QGQQ (SEQ ID NO: 5075), LNRQ (SEQ ID NO: 5076), QNPP (SEQ ID NO: 5077), QNLQ (SEQ ID NO: 5078), QDQE (SEQ ID NO: 5079), QDQQ (SEQ ID NO: 5080), HWQQ (SEQ ID NO: 5081), PNQQ (SEQ ID NO: 5082), PEQQ (SEQ ID NO: 5083), QRTM (SEQ ID NO: 5084), LHQH (SEQ ID NO: 5085), QHRI (SEQ ID NO: 5086), QYIH (SEQ ID NO: 5087), QKFE (SEQ ID NO: 5088), QFPS (SEQ ID NO: 5089), QNPL (SEQ ID NO: 5090), QAIK (SEQ ID NO: 5091), QNRQ (SEQ ID NO: 5092), QYQH (SEQ ID NO: 5093), QNPQ (SEQ ID NO: 5094), QHQL (SEQ ID NO: 5095), QSPP (SEQ ID NO: 5096), QAKL (SEQ ID NO: 5097), KSQQ (SEQ ID NO: 5098), QDRP (SEQ ID NO: 5099), QNLG (SEQ ID NO: 5100), QAFH (SEQ ID NO: 5101), QNAQ (SEQ ID NO: 5102), HNQL (SEQ ID NO: 5103), QKLN (SEQ ID NO: 5104), QNVQ (SEQ ID NO: 5105), QAQQ (SEQ ID NO: 5106), QTPP (SEQ ID NO: 5107), QPPA (SEQ ID NO: 5108), QERP (SEQ ID NO: 5109), QDLQ (SEQ ID NO: 5110), QAMH (SEQ ID NO: 5111), QHPS (SEQ ID NO: 5112), PGLQ (SEQ ID NO: 5113), QGIR (SEQ ID NO: 5114), QAPA (SEQ ID NO: 5115), QIPP (SEQ ID NO: 5116), QTQL (SEQ ID NO: 5117), QAPS (SEQ ID NO: 5118), QNTY (SEQ ID NO: 5119), QDKQ (SEQ ID NO: 5120), QNHL (SEQ ID NO: 5121), QIGM (SEQ ID NO: 5122), LNKQ (SEQ ID NO: 5123), PNQL (SEQ ID NO: 5124), QLQQ (SEQ ID NO: 5125), QRMS (SEQ ID NO: 5126), QGIL (SEQ ID NO: 5127), QDRQ (SEQ ID NO: 5128), RDWQ (SEQ ID NO: 5129), QERS (SEQ ID NO: 5130), QNYQ (SEQ ID NO: 5131), QRTC (SEQ ID NO: 5132), QIGH (SEQ ID NO: 5133), QGAI (SEQ ID NO: 5134), QVPP (SEQ ID NO: 5135), QVQQ (SEQ ID NO: 5136), LMRQ (SEQ ID NO: 5137), QYSV (SEQ ID NO: 5138), QAIT (SEQ ID NO: 5139), QKTL (SEQ ID NO: 5140), QLHH (SEQ ID NO: 5141), QNII (SEQ ID NO: 5142), QGHH (SEQ ID NO: 5143), QSKV (SEQ ID NO: 5144), QLPS (SEQ ID NO: 5145), IGKQ (SEQ ID NO: 5146), QAIH (SEQ ID NO: 5147), QHGL (SEQ ID NO: 5148), QFMC (SEQ ID NO: 5149), QNQM (SEQ ID NO: 5150), QHLQ (SEQ ID NO: 5151), QPAR (SEQ ID NO: 5152), QSLQ (SEQ ID NO: 5153), QSQL (SEQ ID NO: 5154), HSQQ (SEQ ID NO: 5155), QMPS (SEQ ID NO: 5156), QGSL (SEQ ID NO: 5157), QVPA (SEQ ID NO: 5158), HYQQ (SEQ ID NO: 5159), QVPS (SEQ ID NO: 5160), RGEQ (SEQ ID NO: 5161), PGQQ (SEQ ID NO: 5162), LEQQ (SEQ ID NO: 5163), QNQS (SEQ ID NO: 5164), QKVI (SEQ ID NO: 5165), QNND (SEQ ID NO: 5166), QSVH (SEQ ID NO: 5167), QPLG (SEQ ID NO: 5168), HNQE (SEQ ID NO: 5169), QIQQ (SEQ ID NO: 5170), QVRN (SEQ ID NO: 5171), PSNQ (SEQ ID NO: 5172), QVGH (SEQ ID NO: 5173), QRDI (SEQ ID NO: 5174), QMPN (SEQ ID NO: 5175), RGLQ (SEQ ID NO: 5176), PSLQ (SEQ ID NO: 5177), QRDQ (SEQ ID NO: 5178), QAKG (SEQ ID NO: 5179), QSAH (SEQ ID NO: 5180), QSTM (SEQ ID NO: 5181), QREM (SEQ ID NO: 5182), QYRA (SEQ ID NO: 5183), QRQQ (SEQ ID NO: 5184), QWQQ (SEQ ID NO: 5185), QRMN (SEQ ID NO: 5186), GDSQ (SEQ ID NO: 5187), QKIS (SEQ ID NO: 5188), PSMQ (SEQ ID NO: 5189), SPRQ (SEQ ID NO: 5190), MEQQ (SEQ ID NO: 5191), QYQN (SEQ ID NO: 5192), QIRQ (SEQ ID NO: 5193), QSVQ (SEQ ID NO: 5194), RSQQ (SEQ ID NO: 5195), QNKL (SEQ ID NO: 5196), QIQH (SEQ ID NO: 5197), PRQQ (SEQ ID NO: 5198), HTQQ (SEQ ID NO: 5199), QRQH (SEQ ID NO: 5200), RNQE (SEQ ID NO: 5201), QSKQ (SEQ ID NO: 5202), QNQP (SEQ ID NO: 5203), QSPQ (SEQ ID NO: 5204), QTRQ (SEQ ID NO: 5205), QNLH (SEQ ID NO: 5206), QNQE (SEQ ID NO: 5207), LNQP (SEQ ID NO: 5208), QNQD (SEQ ID NO: 5209), QNLL (SEQ ID NO: 5210), QLVI (SEQ ID NO: 5211), RTQE (SEQ ID NO: 5212), QTHQ (SEQ ID NO: 5213), QDQH (SEQ ID NO: 5214), QSQH (SEQ ID NO: 5215), VRQQ (SEQ ID NO: 5216), AWQQ (SEQ ID NO: 5217), QSVP (SEQ ID NO: 5218), QNIQ (SEQ ID NO: 5219), LDQQ (SEQ ID NO: 5220), PDQQ (SEQ ID NO: 5221), ESQQ (SEQ ID NO: 5222), QRQL (SEQ ID NO: 5223), QIIV (SEQ ID NO: 5224), QKQS (SEQ ID NO: 5225), QSHQ (SEQ ID NO: 5226), QFVV (SEQ ID NO: 5227), QSQP (SEQ ID NO: 5228), QNEQ (SEQ ID NO: 5229), INQQ (SEQ ID NO: 5230), RNRQ (SEQ ID NO: 5231), RDQK (SEQ ID NO: 5232), QWKR (SEQ ID NO: 5233), ENRQ (SEQ ID NO: 5234), QTQP (SEQ ID NO: 5235), QKQL (SEQ ID NO: 5236), RNQL (SEQ ID NO: 5237), ISIQ (SEQ ID NO: 5238), QTVC (SEQ ID NO: 5239), QQIM (SEQ ID NO: 5240), LNHQ (SEQ ID NO: 5241), QNQA (SEQ ID NO: 5242), QMIH (SEQ ID NO: 5243), RNHQ (SEQ ID NO: 5244), or QKMN (SEQ ID NO: 5245), or any dipeptide or tripeptide thereof. In some embodiments, [N1]-[N2]-[N3]-[N4] is or comprises: the amino acid sequence of any of SEQ ID NOs: 1800-2241; an amino acid sequence comprising any portion of any of the aforesaid amino acid sequences (e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids, e.g., consecutive amino acids) thereof; an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the aforesaid amino acid sequences; or an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the aforesaid amino acid sequences. In some embodiments, [N1]-[N2]-[N3]-[N4] is or comprises GSGSPHSKAQNQQ (SEQ ID NO: 1801). In some embodiments, [N1]-[N2]-[N3]-[N4] is or comprises GHDSPHKSGQNQQ (SEQ ID NO: 1800).

In some embodiments, the peptide comprising an amino acid sequence having the formula [N1]-[N2]-[N3], further comprises [N0], which comprises XA XB and XC. In some embodiments, XA of [N0] is T, S, Y, M, A, C, I, R, L, D, F, V, Q, N, H, E, or G. In some embodiments, XB of [N0] is I, M, P, E, N, D, S, A, T, G, Q, F, V, L, C, H, R, W, or L. In some embodiments, XC of [N0] is N, M, E, G, Y, W, T, I, Q, F, V, A, L, I, P, K, R, H, S, D, or S. In some embodiments, [N0] is or comprises TIN, SMN, TIM, YLS, GLS, MPE, MEG, MEY, AEW, CEW, ANN, IPE, ADM, IEY, ADY, IET, MEW, CEY, RIN, MEI, LEY, ADW, IEI, DIM, FEQ, MEF, CDQ, LPE, IEN, MES, AEI, VEY, IIN, TSN, IEV, MEM, AEV, MDA, VEW, AEQ, LEW, MEL, MET, MEA, IES, MEV, CEI, ATN, MDG, QEV, ADQ, NMN, IEM, ISN, TGN, QQQ, HDW, IEG, TII, TFP, TEK, EIN, TVN, TFN, SIN, TER, TSY, ELH, AIN, SVN, TDN, TFH, TVH, TEN, TSS, TID, TCN, NIN, TEH, AEM, AIK, TDK, TFK, SDQ, TEI, NTN, TET, SIK, TEL, TEA, TAN, TIY, TFS, TES, TTN, TED, TNN, EVH, TIS, TVR, TDR, TIK, NHI, TIP, ESD, TDL, TVP, TVI, AEH, NCL, TVK, NAD, TIT, NCV, TIR, NAL, VIN, TIQ, TEF, TRE, QGE, SEK, NVN, GGE, EFV, SDK, TEQ, EVQ, TEY, NCW, TDV, SDI, NSI, NSL, EVV, TEP, SEL, TWQ, TEV, AVN, GVL, TLN, TEG, TRD, NAI, AEN, AET, ETA, NNL, or any dipeptide thereof. In some embodiments, [N0]-[N1]-[N2]-[N3]-[N4] is or comprises the amino acid sequence of any one of SEQ ID NOs: 2242-2886; an amino acid sequence comprising any portion of any of the aforesaid amino acid sequences (e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids, e.g., consecutive amino acids) thereof; an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the aforesaid amino acid sequences; or an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the aforesaid amino acid sequences. In some embodiments, [N0]-[N1]-[N2]-[N3]-[N4] is or comprises TINGSGSPHSKAQNQQ (SEQ ID NO: 2242). In some embodiments, [N0]-[N1]-[N2]-[N3]-[N4] is or comprises TINGHDSPHKSGQNQQ (SEQ ID NO: 2243).

In some embodiments, [N3] is present immediately subsequent to [N2]. In some embodiments, the peptide comprises from N-terminus to C-terminus, [N2]-[N3]. In some embodiments, the peptide comprises from N-terminus to C-terminus, [N1]-[N2]-[N3]. In some embodiments, the peptide comprises from N-terminus to C-terminus, [N1]-[N2]-[N3]-[N4]. In some embodiments, the peptide comprises from N-terminus to C-terminus, [N0]-[N1]-[N2]-[N3]. In some embodiments, the peptide comprises from N-terminus to C-terminus, [N0]-[N1]-[N2]-[N3]-[N4].

In some embodiments, the peptide comprises an amino acid sequence having the formula [A][B](SEQ ID NO: 4694), wherein [A] comprises the amino acid sequence of GSGSPH (SEQ ID NO: 4695) and [B] comprises X1 X2 X3 X4 X5 X6 X7. In some embodiments, position X1 of [B] is S, C, F, or V. In some embodiments, position X2 of [B] is K, L, R, I, E, Y, V, or S. In some embodiments, X3 of [B] is A, R, L, G, I, Y, S, F, or W. In some embodiments X4 of [B] is W, Q, R, G, L, V, S, or F. In some embodiments, position X5 of [B] is N, Y, R, C, K, or L. In some embodiments, position X6 of [B] is Q, G, K, R, T, L, or Y. In some embodiment, position X7 of [B] is Q, L, R, or V. In some embodiments, [B] comprises S LLWNQQ (SEQ ID NO: 5247), SKAQYYV (SEQ ID NO: 5248), SKLRRQQ (SEQ ID NO: 5249), SIWQNQQ (SEQ ID NO: 5250), SKAGCGQ (SEQ ID NO: 5251), SRAQNQQ (SEQ ID NO: 5252), SKRLRQQ (SEQ ID NO: 5253), SLRRNQQ (SEQ ID NO: 5254), SRGRNQQ (SEQ ID NO: 5255), SEIVNQQ (SEQ ID NO: 5256), SSRRNQQ (SEQ ID NO: 5257), CLLQNQQ (SEQ ID NO: 5258), SKAFRLQ (SEQ ID NO: 5259), CLAQNQQ (SEQ ID NO: 5260), FLRQNQQ (SEQ ID NO: 5261), SLRFNQQ (SEQ ID NO: 5262), SYLRNQQ (SEQ ID NO: 5263), CSLQNQQ (SEQ ID NO: 5264), VLWQNQQ (SEQ ID NO: 5265), SKWLLQQ (SEQ ID NO: 5266), SLWSNQQ (SEQ ID NO: 5267), SKRRLQQ (SEQ ID NO: 5268), SVYLNQQ (SEQ ID NO: 5269), SLWLNQQ (SEQ ID NO: 5270), SKAQRKL (SEQ ID NO: 5271), SKALRRQ (SEQ ID NO: 5272), SKAQRLR (SEQ ID NO: 5273), SKAQNQQ (SEQ ID NO: 5274), SKAQRRL (SEQ ID NO: 5275), SKARRQQ (SEQ ID NO: 5276), SKARRLQ (SEQ ID NO: 5277), SKSRRQQ (SEQ ID NO: 5278), SKARLRQ (SEQ ID NO: 5279), SKASKRQ (SEQ ID NO: 5280), VRRQNQQ (SEQ ID NO: 5281), SKAQLYR (SEQ ID NO: 5282), SLFRNQQ (SEQ ID NO: 5283), SKAQLTV (SEQ ID NO: 5284), or any dipeptide, tripeptide, tetrapeptide, pentapeptide, or hexapeptide thereof. In some embodiments, [A][B] comprises GSGSPHSLLWNQQ (SEQ ID NO: 5285), GSGSPHSKAQYYV (SEQ ID NO: 2060), GSGSPHSKLRRQQ (SEQ ID NO: 2061), GSGSPHSIWQNQQ (SEQ ID NO: 5286), GSGSPHSKAGCGQ (SEQ ID NO: 2062), GSGSPHSRAQNQQ (SEQ ID NO: 2063), GSGSPHSKRLRQQ (SEQ ID NO: 2064), GSGSPHSLRRNQQ (SEQ ID NO: 2065), GSGSPHSRGRNQQ (SEQ ID NO: 2066), GSGSPHSEIVNQQ (SEQ ID NO: 5287), GSGSPHSSRRNQQ (SEQ ID NO: 2067), GSGSPHCLLQNQQ (SEQ ID NO: 5288), GSGSPHSKAFRLQ (SEQ ID NO: 2068), GSGSPHCLAQNQQ (SEQ ID NO: 5289), GSGSPHFLRQNQQ (SEQ ID NO: 2070), GSGSPHSLRFNQQ (SEQ ID NO: 2071), GSGSPHSYLRNQQ (SEQ ID NO: 5290), GSGSPHCSLQNQQ (SEQ ID NO: 5291), GSGSPHVLWQNQQ (SEQ ID NO: 5292), GSGSPHSKWLLQQ (SEQ ID NO: 2072), GSGSPHSLWSNQQ (SEQ ID NO: 5293), GSGSPHSKRRLQQ (SEQ ID NO: 2073), GSGSPHSVYLNQQ (SEQ ID NO: 5294), GSGSPHSLWLNQQ (SEQ ID NO: 5295), GSGSPHSKAQRKL (SEQ ID NO: 2074), GSGSPHSKALRRQ (SEQ ID NO: 2075), GSGSPHSKAQRLR (SEQ ID NO: 2076), GSGSPHSKAQNQQ (SEQ ID NO: 1801), GSGSPHSKAQRRL (SEQ ID NO: 2077), GSGSPHSKARRQQ (SEQ ID NO: 2078), GSGSPHSKARRLQ (SEQ ID NO: 2079), GSGSPHSKSRRQQ (SEQ ID NO: 2080), GSGSPHSKARLRQ (SEQ ID NO: 2082), GSGSPHSKASKRQ (SEQ ID NO: 2083), GSGSPHVRRQNQQ (SEQ ID NO: 2084), GSGSPHSKAQLYR (SEQ ID NO: 2085), GSGSPHSLFRNQQ (SEQ ID NO: 5296), GSGSPHSKAQLTV (SEQ ID NO: 2086), or any portion thereof, e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids, e.g., consecutive amino acids, thereof. In some embodiments, [B] is present immediately subsequent to [A]. In some embodiments, the peptide comprises from N-terminus to C-terminus, [A][B].

In some embodiments, the peptide comprises an amino acid sequence having the formula [A][B](SEQ ID NO: 4699), wherein [A] comprises X1 X2 X3 X4 X5 X6 and [B] comprises SPHKSG (SEQ ID NO: 946). In some embodiments, position X1 of [A] is T, M, A, C, I, R, L, D, F, V, Q, N, or H. In some embodiments, position X2 of [A] is I, P, E, N, D, S, A, T, M, or Q. In some embodiments, position X3 of [A] is N, E, G, Y, W, M, T, I, K, Q, F, S, V, A, or L. In some embodiments, position X4 of [A] is G, D, R, or E. In some embodiments, position X5 of [A] is H, Q, N, or D. In some embodiments, position X6 of [A] is D or R. In some embodiments, [A] comprises TINGHD (SEQ ID NO: 5297), MPEGHD (SEQ ID NO: 5298), MEGGHD (SEQ ID NO: 5299), MEYGHD (SEQ ID NO: 5300), AEWGHD (SEQ ID NO: 5301), CEWGHD (SEQ ID NO: 5302), ANNGQD (SEQ ID NO: 5303), IPEGHD (SEQ ID NO: 5304), ADMGHD (SEQ ID NO: 5305), IEYGHD (SEQ ID NO: 5306), ADYGHD (SEQ ID NO: 5307), IETGHD (SEQ ID NO: 5308), MEWGHD (SEQ ID NO: 5309), CEYGHD (SEQ ID NO: 5310), RINGHD (SEQ ID NO: 5311), MEIGHD (SEQ ID NO: 5312), LEYGHD (SEQ ID NO: 5313), ADWGHD (SEQ ID NO: 5314), IEIGHD (SEQ ID NO: 5315), TIKDND (SEQ ID NO: 5316), DIMGHD (SEQ ID NO: 5317), FEQGHD (SEQ ID NO: 5318), MEFGHD (SEQ ID NO: 5319), CDQGHD (SEQ ID NO: 5320), LPEGHD (SEQ ID NO: 5321), IENGHD (SEQ ID NO: 5322), MESGHD (SEQ ID NO: 5323), AEIGHD (SEQ ID NO: 5324), VEYGHD (SEQ ID NO: 5325), TSNGDD (SEQ ID NO: 5326), IEVGHD (SEQ ID NO: 5327), MEMGHD (SEQ ID NO: 5328), AEVGHD (SEQ ID NO: 5329), MDAGHD (SEQ ID NO: 5330), VEWGHD (SEQ ID NO: 5331), AEQGHD (SEQ ID NO: 5332), LEWGHD (SEQ ID NO: 5333), MELGHD (SEQ ID NO: 5334), METGHD (SEQ ID NO: 5335), MEAGHD (SEQ ID NO: 5336), TINRQR (SEQ ID NO: 5337), IESGHD (SEQ ID NO: 5338), TAKDHD (SEQ ID NO: 5339), MEVGHD (SEQ ID NO: 5340), CEIGHD (SEQ ID NO: 5341), ATNGHD (SEQ ID NO: 5342), MDGGHD (SEQ ID NO: 5343), QEVGHD (SEQ ID NO: 5344), ADQGHD (SEQ ID NO: 5345), NMNGHD (SEQ ID NO: 5346), TPWEHD (SEQ ID NO: 5347), IEMGHD (SEQ ID NO: 5348), TANEHD (SEQ ID NO: 5349), QQQGHD (SEQ ID NO: 5350), TPQDHD (SEQ ID NO: 5351), HDWGHD (SEQ ID NO: 5352), IEGGHD (SEQ ID NO: 5353), or any dipeptide, tripeptide, tetrapeptide, or pentapeptide thereof. In some embodiments, [A][B] comprises TINGHDSPHKR (SEQ ID NO: 5354), MPEGHDSPHKS (SEQ ID NO: 5355), MEGGHDSPHKS (SEQ ID NO: 5356), MEYGHDSPHKS (SEQ ID NO: 5357), AEWGHDSPHKS (SEQ ID NO: 5358), CEWGHDSPHKS (SEQ ID NO: 5359), ANNGQDSPHKS (SEQ ID NO: 5360), IPEGHDSPHKS (SEQ ID NO: 5361), ADMGHDSPHKS (SEQ ID NO: 5362), IEYGHDSPHKS (SEQ ID NO: 5363), ADYGHDSPHKS (SEQ ID NO: 5364), IETGHDSPHKS (SEQ ID NO: 5365), MEWGHDSPHKS (SEQ ID NO: 5366), CEYGHDSPHKS (SEQ ID NO: 5367), RINGHDSPHKS (SEQ ID NO: 5368), MEIGHDSPHKS (SEQ ID NO: 5369), LEYGHDSPHKS (SEQ ID NO: 5370), ADWGHDSPHKS (SEQ ID NO: 5371), IEIGHDSPHKS (SEQ ID NO: 5372), TIKDNDSPHKS (SEQ ID NO: 5373), DIMGHDSPHKS (SEQ ID NO: 5374), FEQGHDSPHKS (SEQ ID NO: 5375), MEFGHDSPHKS (SEQ ID NO: 5376), CDQGHDSPHKS (SEQ ID NO: 5377), LPEGHDSPHKS (SEQ ID NO: 5378), IENGHDSPHKS (SEQ ID NO: 5379), MESGHDSPHKS (SEQ ID NO: 5380), AEIGHDSPHKS (SEQ ID NO: 5381), VEYGHDSPHKS (SEQ ID NO: 5382), TSNGDDSPHKS (SEQ ID NO: 5383), IEVGHDSPHKS (SEQ ID NO: 5384), MEMGHDSPHKS (SEQ ID NO: 5385), AEVGHDSPHKS (SEQ ID NO: 5386), MDAGHDSPHKS (SEQ ID NO: 5387), VEWGHDSPHKS (SEQ ID NO: 5388), AEQGHDSPHKS (SEQ ID NO: 5389), LEWGHDSPHKS (SEQ ID NO: 5390), MELGHDSPHKS (SEQ ID NO: 5391), METGHDSPHKS (SEQ ID NO: 5392), MEAGHDSPHKS (SEQ ID NO: 5393), TINRQRSPHKS (SEQ ID NO: 5394), IESGHDSPHKS (SEQ ID NO: 5395), TAKDHDSPHKS (SEQ ID NO: 5396), MEVGHDSPHKS (SEQ ID NO: 5397), CEIGHDSPHKS (SEQ ID NO: 5398), ATNGHDSPHKS (SEQ ID NO: 5399), MDGGHDSPHKS (SEQ ID NO: 5400), QEVGHDSPHKS (SEQ ID NO: 5401), ADQGHDSPHKS (SEQ ID NO: 5402), NMNGHDSPHKS (SEQ ID NO: 5403), TPWEHDSPHKS (SEQ ID NO: 5404), IEMGHDSPHKS (SEQ ID NO: 5405), TANEHDSPHKS (SEQ ID NO: 5406), TINGHDSPHKS (SEQ ID NO: 5407), QQQGHDSPHKS (SEQ ID NO: 5408), TPQDHDSPHKS (SEQ ID NO: 5409), HDWGHDSPHKS (SEQ ID NO: 5410), IEGGHDSPHKS (SEQ ID NO: 5411), or any portion thereof, e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids, e.g., consecutive amino acids, thereof. In some embodiments, [B] is present immediately subsequent to [A]. In some embodiments, the peptide comprises from N-terminus to C-terminus, [A][B].

In some embodiments, the peptide comprises an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 consecutive amino acids from any one of the sequences provided in Tables 1, 2A, 2B, 13-19. In some embodiments, the peptide comprises an amino acid sequence comprising at least 3, 4, or 5 consecutive amino acids from any one of SEQ ID NOs: 945-980 or 985-986. In some embodiments, the peptide comprises an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 consecutive amino acids from any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909.

In some embodiments, the 3 consecutive amino acids comprise SPH. In some embodiments, the 4 consecutive amino acids comprise SPHS (SEQ ID NO: 4700). In some embodiments, the 5 consecutive amino acids comprise SPHSK (SEQ ID NO: 4701). In some embodiments, the 6 consecutive amino acids comprise SPHSKA (SEQ ID NO: 941).

In some embodiments, 3 consecutive amino acids comprise HDS. In some embodiments, the 4 consecutive amino acids comprise HDSP (SEQ ID NO: 4702). In some embodiments, the 5 consecutive amino acids comprise HDSPH (SEQ ID NO: 4703). In some embodiments, the 6 consecutive amino acids comprise HDSPHK (SEQ ID NO: 2).

In some embodiments, the 3 consecutive amino acids comprise SPH. In some embodiments, the 4 consecutive amino acids comprise SPHK (SEQ ID NO: 6398). In some embodiments, the 5 consecutive amino acids comprise SPHKY (SEQ ID NO: 4715). In some embodiments, the 6 consecutive amino acids comprise SPHKYG (SEQ ID NO: 966).

In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of any one of the sequences provided in Tables 1, 2A, 2B, 13-19. In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids, relative to the amino acid sequence of any one of the sequences provided in Tables 1, 2A, 2B, 13-19. In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of any one of SEQ ID NOs: 945-980 or 985-986. In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids, relative to the amino acid sequence of any one of SEQ ID NOs: 945-980 or 985-986. In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909. In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909. In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of SEQ ID NO: 3589. In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of SEQ ID NO: 3589. In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of SEQ ID NO: 1754. In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of SEQ ID NO: 1754.

In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941). In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941).

In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2). In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2).

In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of SPHKYG (SEQ ID NO: 966). In some embodiments, the peptide comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids relative to the amino acid sequence of SPHKYG (SEQ ID NO: 966).

In some embodiments, the peptide comprises the amino acid sequence of any of the sequences provided in Tables 1, 2A, 2B, 13-19. In some embodiments, the peptide comprises the amino acid sequence of any of SEQ ID NOs: 945-980 or 985-986. In some embodiments, the peptide comprises the amino acid sequence of any of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909. In some embodiments, the peptide comprises the amino acid sequence of SEQ ID NO: 941. In some embodiments, the peptide comprises the amino acid sequence of SEQ ID NO: 943. In some embodiments, the peptide comprises the amino acid sequence of SEQ ID NO: 2. In some embodiments, the peptide comprises the amino acid sequence of SEQ ID NO: 3589. In some embodiments, the peptide comprises the amino acid sequence of SEQ ID NO: 1754. In some embodiments, the peptide comprises the amino acid sequence of SEQ ID NO: 3241. In some embodiments, the peptide comprises the amino acid sequence of SEQ ID NO: 4100. In some embodiments, the peptide comprises the amino acid sequence of SEQ ID NO: 4062. In some embodiments, the peptide comprises the amino acid sequence of SEQ ID NO: 4486.

In some embodiments, the peptide comprises an amino acid sequence encoded by a nucleotide sequence described herein, e.g., a nucleotide sequence of Table 2A. In some embodiments, the peptide comprises an amino acid sequence encoded by a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 942. In some embodiments, the peptide comprises an amino acid sequence encoded by a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides, relative to the nucleotide sequence of SEQ ID NO: 942. In some embodiments, the peptide comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 942, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto. In some embodiments, the peptide comprises an amino acid sequence encoded by a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 944. In some embodiments, the peptide comprises an amino acid sequence encoded by a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides, relative to the nucleotide sequence of SEQ ID NO: 944. In some embodiments, the peptide comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 944, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto.

In some embodiments, the nucleotide sequence encoding a peptide described herein comprises a nucleotide sequence described herein, e.g., as described in Table 2A. In some embodiments, the nucleotide sequence encoding a peptide described herein is codon optimized. In some embodiments, the nucleotide sequence encoding a peptide described herein is isolated, e.g., recombinant.

In some embodiments the nucleotide sequence encoding a peptide described herein comprises the nucleotide sequence of SEQ ID NO: 942, or a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 942. In some embodiments, the nucleotide sequence encoding a peptide described herein comprises a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides, relative to the nucleotide sequence of SEQ ID NO: 942. In some embodiments the nucleic acid sequence encoding a peptide described herein comprises a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 942, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto.

In some embodiments, the nucleic acid encoding a peptide described herein comprises the nucleotide sequence of SEQ ID NO: 3, or a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 3. In some embodiments, the nucleotide sequence encoding a peptide described herein comprises a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides, relative to the nucleotide sequence of SEQ ID NO: 3. In some embodiments the nucleic acid encoding a peptide described herein comprises a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 3, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto.

In some embodiments, the nucleic acid encoding a peptide described herein comprises the nucleotide sequence of SEQ ID NO: 944, or a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 944. In some embodiments, the nucleotide sequence encoding a peptide described herein comprises a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides, relative to the nucleotide sequence of SEQ ID NO: 944. In some embodiments the nucleic acid encoding a peptide described herein comprises a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 944, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto.

In some embodiments, a peptide described herein is fused or coupled, e.g., conjugated, to an active agent. In some embodiments, the active agent is a therapeutic agent. In some embodiments, the agent is a therapeutic agent. In some embodiments, the active agent comprises a therapeutic protein, an antibody molecule, an enzyme, one or more components of a genome editing system, an Fc polypeptide fused or coupled (e.g., covalently or non covalently) to a therapeutic agent, and/or an RNAi agent (e.g., a dsRNA, antisense oligonucleotide (ASO), siRNA, shRNA, pre-miRNA, pri-miRNA, miRNA, stRNA, lncRNA, piRNA, or snoRNA). In some embodiments, the therapeutic agent is an antibody. In some embodiments, the peptide is fused or coupled, e.g., conjugated (e.g., directly or indirectly) to the Fc region of the antibody, e.g., at the C-terminus of the Fc region or the N-terminus of the Fc region. In some embodiments, the therapeutic agent is an RNAi agent. In some embodiments, the RNAi agent is a siRNA or an ASO. In some embodiments, the ASO or siRNA comprises at least one (e.g., one or more or all) modified nucleotides. In some embodiments, the peptide is fused or coupled, e.g., conjugated (e.g., directly or indirectly via a linker), to at least one strand of the RNAi agent. In some embodiments, the peptide is conjugated, e.g., directly or indirectly via a linker, to the C-terminus of at least one strand of the RNAi agent. In some embodiments, the peptide is conjugated, e.g., directly or indirectly via a linker, to an internal nucleotide of at least one strand of the RNAi agent. In some embodiments, the at least one strand is the sense strand. In some embodiments, the therapeutic agent modulates, e.g., inhibits, decreases, or increases, expression of, a CNS related gene, mRNA, and/or protein.

In some embodiments, the active agent is a diagnostic agent. In some embodiments, the diagnostic agent is or comprises an imaging agent (e.g., a protein or small molecule compound coupled to a detectable moiety). In some embodiments, the imaging agent comprises a PET or MRI ligand, or an antibody molecule coupled to a detectable moiety. In some embodiments, the detectable moiety is or comprises a radiolabel, a fluorophore, a chromophore, or an affinity tag. In some embodiments, the radiolabel is or comprises tc99m, iodine-123, a spin label, iodine-131, indium-111, fluorine-19, carbon-13, nitrogen-15, oxygen-17, gadolinium, manganese, or iron. In some embodiments, the active agent is a small molecule. In some embodiments, the active agent is a ribonucleic acid complex (e.g., a Cas9/gRNA complex), a plasmid, a closed-end DNA, a circ-RNA, or an mRNA.

In some embodiments, at least 1-5, e.g., at least 1, 2, 3, 4, or 5, peptides are fused or coupled, e.g., conjugated, to an active agent, e.g., a therapeutic agent or a diagnostic agent. In some embodiments, the at least 1-5, e.g., at least 1, 2, 3, 4, or 5, peptides comprise the same amino acid sequence. In some embodiments, the at least 1-5, e.g., at least 1, 2, 3, 4, or 5, peptides comprise different amino acid sequences. In some embodiments, the at least 1-5, e.g., at least 1, 2, 3, 4, or 5, peptides are present in tandem (e.g., connected directly or indirectly via a linker) or in a multimeric configuration. In some embodiments, the peptide comprises an amino acid sequence of at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 20, 25, 30, or 35 amino acids in length.

In some embodiments, the peptide covalently linked, e.g., directly or indirectly via a linker, to the active agent. In some embodiments, the peptide is conjugated to the active agent via a linker. In some embodiments, the linker is a cleavable linker or a non-cleavable linker. In some embodiments, the cleavable linker is a pH sensitive linker or an enzyme sensitive linker. In some embodiments, the pH sensitive linker comprises a hydrazine/hydrazone linker or a disulfide linker. In some embodiments, the enzyme sensitive linker comprises a peptide based linker, e.g., a peptide linker sensitive to a protease (e.g., a lysosomal protease); or a beta-glucuronide linker. In some embodiments, the non-cleavable linker is a linker comprising a thioether group or a maleimidocaproyl group. In some embodiments, the peptide and the active agent are fused or coupled post-translationally, e.g., using click chemistry. In some embodiments, the peptide and the active agent are fused or couple via chemically induced dimerization. In some embodiments, the peptide is present N-terminal relative to the active agent. In some embodiments, the peptide is present C-terminal relative to the active agent.

In some embodiments, the peptide is present or coupled to a carrier. In some embodiments, the carrier comprises an exosome, a microvesicle, or a lipid nanoparticle (LNP). In some embodiments, the carrier comprises a therapeutic agent (e.g., an RNAi agent (e.g., an dsRNA, a siRNA, a shRNA, a pre-miRNA, a pri-miRNA, a miRNA, a stRNA, a lncRNA, a piRNA, an antisense oligonucleotide agent (ASO), or a snoRNA), an mRNA, a ribonucleoprotein complex (e.g., a Cas9/gRNA complex), or a circRNA). In some embodiments, the peptide is present on the surface of the carrier. In some embodiments, at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, or 80% of the surface of the carrier comprises at least 1-5, e.g., at least 1, 2, 3, 4, or 5 peptides described herein.

The present disclosure also provides a nucleic acid or polynucleotide encoding any of the peptides described herein and AAV capsid variants, AAV particles, vectors, and cells comprising the same.

AAV Capsid Variant

In some embodiments, an AAV particle described herein comprises an AAV capsid variant, e.g., an AAV capsid variant described herein (e.g., an AAV capsid variant comprising a peptide described herein). In some embodiments, an AAV capsid variant comprises a peptide as set forth in any of Tables 1, 2A, 2B, 13-19.

In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence having the formula [N1]-[N2]-[N3], wherein [N2] comprises the amino acid sequence of SPH and [N3] comprises X4, X5, and X6, wherein at least one of X4, X5, or X6 is a basic amino acid, e.g., a K or R. In some embodiments, position X4 of [N2] is K. In some embodiments, position X5 of [N2] is K.

In some embodiments, [N1] comprises X1, X2, and X3, wherein at least one of X1, X2, or X3 is G. In some embodiments, position X1 of [N1] is G, V, R, D, E, M, T, I, S, A, N, L, K, H, P, W, or C. In some embodiments, position X2 of [N1] is: S, V, L, N, D, H, R, P, G, T, I, A, E, Y, M, or Q. In some embodiments, position X3 of [N1] is: G, C, L, D, E, Y, H, V, A, N, P, or S. In some embodiments, [N1] comprises GS, SG, GH, HD, GQ, QD, VS, CS, GR, RG, QS, SH, MS, RN, TS, IS, GP, ES, SS, GN, AS, NS, LS, GG, KS, GT, PS, RS, GI, WS, DS, ID, GL, DA, DG, ME, EN, KN, KE, AI, NG, PG, TG, SV, IG, LG, AG, EG, SA, YD, HE, HG, RD, ND, PD, MG, QV, DD, HN, HP, GY, GM, GD, or HS. In some embodiments, [N1] comprises GS, SG, GH, or HD. In some embodiments [N1] is or comprises GSG, GHD, GQD, VSG, CSG, CSH, GQS, GRG, GSH, RVG, GSC, GLL, GDD, GHE, GNY, MSG, RNG, TSG, ISG, GPG, ESG, SSG, GNG, ASG, NSG, LSG, GGG, KSG, HSG, GTG, PSG, GSV, RSG, GIG, WSG, DSG, IDG, GLG, DAG, DGG, MEG, ENG, GSA, KNG, KEG, AIG, GYD, GHG, GRD, GND, GPD, GMG, GQV, GHN, GHP, or GHS. In some embodiments, [N1] is or comprises GSG. In some embodiments, [N1] is or comprises GHD. In some embodiments, [N1]-[N2] comprises SGSPH (SEQ ID NO: 4752), HDSPH (SEQ ID NO: 4703), QDSPH (SEQ ID NO: 4753), RGSPH (SEQ ID NO: 4754), SHSPH (SEQ ID NO: 4755), QSSPH (SEQ ID NO: 4756), DDSPH (SEQ ID NO: 4757), HESPH (SEQ ID NO: 4758), NYSPH (SEQ ID NO: 4759), VGSPH (SEQ ID NO: 4760), SCSPH (SEQ ID NO: 4761), LLSPH (SEQ ID NO: 4762), NGSPH (SEQ ID NO: 4763), PGSPH (SEQ ID NO: 4764), GGSPH (SEQ ID NO: 4765), TGSPH (SEQ ID NO: 4766), SVSPH (SEQ ID NO: 4767), IGSPH (SEQ ID NO: 4768), DGSPH (SEQ ID NO: 4769), LGSPH (SEQ ID NO: 4770), AGSPH (SEQ ID NO: 4771), EGSPH (SEQ ID NO: 4772), SASPH (SEQ ID NO: 4773), YDSPH (SEQ ID NO: 4774), HGSPH (SEQ ID NO: 4775), RDSPH (SEQ ID NO: 4776), NDSPH (SEQ ID NO: 4777), PDSPH (SEQ ID NO: 4778), MGSPH (SEQ ID NO: 4779), QVSPH (SEQ ID NO: 4780), HNSPH (SEQ ID NO: 4781), HPSPH (SEQ ID NO: 4782), or HSSPH (SEQ ID NO: 4783); an amino acid sequence comprising any portion of any of the aforesaid amino acid sequences (e.g., any 2, 3, or 4 amino acids, e.g., consecutive amino acids) thereof; an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the aforesaid amino acid sequences; or an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the aforesaid amino acid sequences. In some embodiments, [N1]-[N2] is or comprises GSGSPH (SEQ ID NO: 4695), GHDSPH (SEQ ID NO: 4784), GQDSPH (SEQ ID NO: 4785), VSGSPH (SEQ ID NO: 4786), CSGSPH (SEQ ID NO: 4787), GRGSPH (SEQ ID NO: 4788), CSHSPH (SEQ ID NO: 4789), GQSSPH (SEQ ID NO: 4790), GSHSPH (SEQ ID NO: 4791), GDDSPH (SEQ ID NO: 4792), GHESPH (SEQ ID NO: 4793), GNYSPH (SEQ ID NO: 4794), RVGSPH (SEQ ID NO: 4795), GSCSPH (SEQ ID NO: 4796), GLLSPH (SEQ ID NO: 4797), MSGSPH (SEQ ID NO: 4798), RNGSPH (SEQ ID NO: 4799), TSGSPH (SEQ ID NO: 4800), ISGSPH (SEQ ID NO: 4801), GPGSPH (SEQ ID NO: 4802), ESGSPH (SEQ ID NO: 4803), SSGSPH (SEQ ID NO: 4804), GNGSPH (SEQ ID NO: 4805), ASGSPH (SEQ ID NO: 4806), NSGSPH (SEQ ID NO: 4807), LSGSPH (SEQ ID NO: 4808), GGGSPH (SEQ ID NO: 4809), KSGSPH (SEQ ID NO: 4810), HSGSPH (SEQ ID NO: 4811), GTGSPH (SEQ ID NO: 4812), PSGSPH (SEQ ID NO: 4813), GSVSPH (SEQ ID NO: 4814), RSGSPH (SEQ ID NO: 4815), GIGSPH (SEQ ID NO: 4816), WSGSPH (SEQ ID NO: 4817), DSGSPH (SEQ ID NO: 4818), IDGSPH (SEQ ID NO: 4819), GLGSPH (SEQ ID NO: 4820), DAGSPH (SEQ ID NO: 4821), DGGSPH (SEQ ID NO: 4822), MEGSPH (SEQ ID NO: 4823), ENGSPH (SEQ ID NO: 4824), GSASPH (SEQ ID NO: 4825), KNGSPH (SEQ ID NO: 4826), KEGSPH (SEQ ID NO: 4827), AIGSPH (SEQ ID NO: 4828), GYDSPH (SEQ ID NO: 4829), GHGSPH (SEQ ID NO: 4830), GRDSPH (SEQ ID NO: 4831), GNDSPH (SEQ ID NO: 4832), GPDSPH (SEQ ID NO: 4833), GMGSPH (SEQ ID NO: 4834), GQVSPH (SEQ ID NO: 4835), GHNSPH (SEQ ID NO: 4836), GHPSPH (SEQ ID NO: 4837), or GHSSPH (SEQ ID NO: 4838); an amino acid sequence comprising any portion of any of the aforesaid amino acid sequences (e.g., any 2, 3, 4, or 5 amino acids, e.g., consecutive amino acids) thereof; an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the aforesaid amino acid sequences; or an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the aforesaid amino acid sequences. In some embodiments, [N1]-[N2] is or comprises GSGSPH (SEQ ID NO: 4695). In some embodiments, [N1]-[N2] is or comprises GHDSPH (SEQ ID NO: 4784).

In some embodiments, X4, X5, or both of [N3] are K. In some embodiments, X4, X5, or X6 of [N3] is R. In some embodiments, position X4 of [N3] is: A, K, V, S, T, G, F, W, V, N, or R. In some embodiments, position X5 of [N3] is: S, K, T, F, I, L, Y, H, M, or R. In some embodiments, position X6 of [N3] is: G, R, A, M, I, N, T, Y, D, P, V, L, E, W, N, Q, K, or S. In some embodiments, [N3] comprises SK, KA, KS, AR, RM, VK, AS, SR, VK, KR, KK, KN, VR, RS, RK, KT, TS, KF, FG, KI, IG, KL, LG, TT, TY, KY, YG, KD, KP, TR, RG, VR, GA, SL, SS, FL, WK, SA, RA, LR, KW, RR, GK, TK, NK, AK, KV, KG, KH, KM, TG, SE, SV, SW, SN, HG, SQ, LW, MG, MA, or SG. In some embodiments, [N3] comprises SK, KA, KS, or SG. In some embodiments, [N3] is or comprises SKA, KSG, ARM, VKS, ASR, VKI, KKN, VRM, RKA, KTS, KFG, KIG, KLG, KTT, KTY, KYG, SKD, SKP, TRG, VRG, KRG, GAR, KSA, KSR, SKL, SRA, SKR, SLR, SRG, SSR, FLR, SKW, SKS, WKA, VRR, SKV, SKT, SKG, GKA, TKA, NKA, SKL, SKN, AKA, KTG, KSL, KSE, KSV, KSW, KSN, KHG, KSQ, KSK, KLW, WKG, KMG, KMA, or RSG. In some embodiments, [N3] is or comprises SKA. In some embodiments, [N3] is or comprises KSG. In some embodiments, [N2]-[N3] comprises SPHSK (SEQ ID NO: 4701), SPHKS (SEQ ID NO: 4704), SPHAR (SEQ ID NO: 4705), SPHVK (SEQ ID NO: 4706), SPHAS (SEQ ID NO: 4707), SPHKK (SEQ ID NO: 4708), SPHVR (SEQ ID NO: 4709), SPHRK (SEQ ID NO: 4710), SPHKT (SEQ ID NO: 4711), SPHKF (SEQ ID NO: 4712), SPHKI (SEQ ID NO: 4713), SPHKL (SEQ ID NO: 4714), SPHKY (SEQ ID NO: 4715), SPHTR (SEQ ID NO: 4716), SPHKR (SEQ ID NO: 4717), SPHGA (SEQ ID NO: 4718), SPHSR (SEQ ID NO: 4719), SPHSL (SEQ ID NO: 4720), SPHSS (SEQ ID NO: 4721), SPHFL (SEQ ID NO: 4722), SPHWK (SEQ ID NO: 4723), SPHGK (SEQ ID NO: 4724), SPHTK (SEQ ID NO: 4725), SPHNK (SEQ ID NO: 4726), SPHAK (SEQ ID NO: 4727), SPHKH (SEQ ID NO: 4728), SPHKM (SEQ ID NO: 4729), or SPHRS (SEQ ID NO: 4730). In some embodiments [N2]-[N3] comprises SPHSK (SEQ ID NO: 4701) or SPHKS (SEQ ID NO: 4704). In some embodiments, [N2]-[N3] is or comprises SPHSKA (SEQ ID NO: 941), SPHKSG (SEQ ID NO: 946), SPHARM (SEQ ID NO: 947), SPHVKS (SEQ ID NO: 948), SPHASR (SEQ ID NO: 949), SPHVKI (SEQ ID NO: 950), SPHKKN (SEQ ID NO: 954), SPHVRM (SEQ ID NO: 955), SPHRKA (SEQ ID NO: 956), SPHKFG (SEQ ID NO: 957), SPHKIG (SEQ ID NO: 958), SPHKLG (SEQ ID NO: 959), SPHKTS (SEQ ID NO: 963), SPHKTT (SEQ ID NO: 964), SPHKTY (SEQ ID NO: 965), SPHKYG (SEQ ID NO: 966), SPHSKD (SEQ ID NO: 967), SPHSKP (SEQ ID NO: 968), SPHTRG (SEQ ID NO: 972), SPHVRG (SEQ ID NO: 973), SPHKRG (SEQ ID NO: 974), SPHGAR (SEQ ID NO: 975), SPHKSA (SEQ ID NO: 977), SPHKSR (SEQ ID NO: 951), SPHSKL (SEQ ID NO: 960), SPHSRA (SEQ ID NO: 969), SPHSKR (SEQ ID NO: 978), SPHSLR (SEQ ID NO: 952), SPHSRG (SEQ ID NO: 961), SPHSSR (SEQ ID NO: 970), SPHFLR (SEQ ID NO: 979), SPHSKW (SEQ ID NO: 953), SPHSKS (SEQ ID NO: 962), SPHWKA (SEQ ID NO: 971), SPHVRR (SEQ ID NO: 980), SPHSKT (SEQ ID NO: 4731), SPHSKG (SEQ ID NO: 4732), SPHGKA (SEQ ID NO: 4733), SPHNKA (SEQ ID NO: 4734), SPHSKN (SEQ ID NO: 4735), SPHAKA (SEQ ID NO: 4736), SPHSKV (SEQ ID NO: 4737), SPHKTG (SEQ ID NO: 4738), SPHTKA (SEQ ID NO: 4739), SPHKSL (SEQ ID NO: 4740), SPHKSE (SEQ ID NO: 4741), SPHKSV (SEQ ID NO: 4742), SPHKSW (SEQ ID NO: 4743), SPHKSN (SEQ ID NO: 4744), SPHKHG (SEQ ID NO: 4745), SPHKSQ (SEQ ID NO: 4746), SPHKSK (SEQ ID NO: 4747), SPHKLW (SEQ ID NO: 4748), SPHWKG (SEQ ID NO: 4749), SPHKMG (SEQ ID NO: 4750), SPHKMA (SEQ ID NO: 4751), or SPHRSG (SEQ ID NO: 976). In some embodiments, [N2]-[N3] is SPHSKA (SEQ ID NO: 941). In some embodiments, [N2]-[N3] is or comprises SPHKSG (SEQ ID NO: 946).

In some embodiments, [N1]-[N2]-[N3] comprises SGSPHSK (SEQ ID NO: 4839), HDSPHKS (SEQ ID NO: 4840), SGSPHAR (SEQ ID NO: 4841), SGSPHVK (SEQ ID NO: 4842), QDSPHKS (SEQ ID NO: 4843), SGSPHKK (SEQ ID NO: 4844), SGSPHVR (SEQ ID NO: 4845), SGSPHAS (SEQ ID NO: 4846), SGSPHRK (SEQ ID NO: 4847), SGSPHKT (SEQ ID NO: 4848), SHSPHKS (SEQ ID NO: 4849), QSSPHRS (SEQ ID NO: 4850), RGSPHAS (SEQ ID NO: 4851), RGSPHSK (SEQ ID NO: 4852), SGSPHKF (SEQ ID NO: 4853), SGSPHKI (SEQ ID NO: 4854), SGSPHKL (SEQ ID NO: 4855), SGSPHKY (SEQ ID NO: 4856), SGSPHTR (SEQ ID NO: 4857), SHSPHKR (SEQ ID NO: 4858), SGSPHGA (SEQ ID NO: 4859), HDSPHKR (SEQ ID NO: 4860), DDSPHKS (SEQ ID NO: 4861), HESPHKS (SEQ ID NO: 4862), NYSPHKI (SEQ ID NO: 4863), SGSPHSR (SEQ ID NO: 4864), SGSPHSL (SEQ ID NO: 4865), SGSPHSS (SEQ ID NO: 4866), VGSPHSK (SEQ ID NO: 4867), SCSPHRK (SEQ ID NO: 4868), SGSPHFL (SEQ ID NO: 4869), LLSPHWK (SEQ ID NO: 4870), NGSPHSK (SEQ ID NO: 4871), PGSPHSK (SEQ ID NO: 4872), GGSPHSK (SEQ ID NO: 4873), TGSPHSK (SEQ ID NO: 4874), SVSPHGK (SEQ ID NO: 4875), SGSPHTK (SEQ ID NO: 4876), IGSPHSK (SEQ ID NO: 4877), DGSPHSK (SEQ ID NO: 4878), SGSPHNK (SEQ ID NO: 4879), LGSPHSK (SEQ ID NO: 4880), AGSPHSK (SEQ ID NO: 4881), EGSPHSK (SEQ ID NO: 4882), SASPHSK (SEQ ID NO: 4883), SGSPHAK (SEQ ID NO: 4884), HDSPHKI (SEQ ID NO: 4885), YDSPHKS (SEQ ID NO: 4886), HDSPHKT (SEQ ID NO: 4887), RGSPHKR (SEQ ID NO: 4888), HGSPHSK (SEQ ID NO: 4889), RDSPHKS (SEQ ID NO: 4890), NDSPHKS (SEQ ID NO: 4891), QDSPHKI (SEQ ID NO: 4892), PDSPHKI (SEQ ID NO: 4893), PDSPHKS (SEQ ID NO: 4894), MGSPHSK (SEQ ID NO: 4895), HDSPHKH (SEQ ID NO: 4896), QVSPHKS (SEQ ID NO: 4897), HNSPHKS (SEQ ID NO: 4898), NGSPHKR (SEQ ID NO: 4899), HDSPHKY (SEQ ID NO: 4900), NDSPHKI (SEQ ID NO: 4901), HDSPHKL (SEQ ID NO: 4902), HPSPHWK (SEQ ID NO: 4903), HDSPHKM (SEQ ID NO: 4904), or HSSPHRS (SEQ ID NO: 4905). In some embodiments, [N1]-[N2]-[N3] is GSGSPHSKA (SEQ ID NO: 4697), GHDSPHKSG (SEQ ID NO: 4698), GSGSPHARM (SEQ ID NO: 4906), GSGSPHVKS (SEQ ID NO: 4907), GQDSPHKSG (SEQ ID NO: 4908), GSGSPHASR (SEQ ID NO: 4909), GSGSPHVKI (SEQ ID NO: 4910), GSGSPHKKN (SEQ ID NO: 4911), GSGSPHVRM (SEQ ID NO: 4912), VSGSPHSKA (SEQ ID NO: 4913), CSGSPHSKA (SEQ ID NO: 4914), GSGSPHRKA (SEQ ID NO: 4915), CSGSPHKTS (SEQ ID NO: 4916), CSHSPHKSG (SEQ ID NO: 4917), GQSSPHRSG (SEQ ID NO: 4918), GRGSPHASR (SEQ ID NO: 4919), GRGSPHSKA (SEQ ID NO: 4920), GSGSPHKFG (SEQ ID NO: 4921), GSGSPHKIG (SEQ ID NO: 4922), GSGSPHKLG (SEQ ID NO: 4923), GSGSPHKTS (SEQ ID NO: 4924), GSGSPHKTT (SEQ ID NO: 4925), GSGSPHKTY (SEQ ID NO: 4926), GSGSPHKYG (SEQ ID NO: 4927), GSGSPHSKD (SEQ ID NO: 4928), GSGSPHSKP (SEQ ID NO: 4929), GSGSPHTRG (SEQ ID NO: 4930), GSGSPHVRG (SEQ ID NO: 4931), GSHSPHKRG (SEQ ID NO: 4932), GSHSPHKSG (SEQ ID NO: 4933), VSGSPHASR (SEQ ID NO: 4934), VSGSPHGAR (SEQ ID NO: 4935), VSGSPHKFG (SEQ ID NO: 4936), GHDSPHKRG (SEQ ID NO: 4937), GDDSPHKSG (SEQ ID NO: 4938), GHESPHKSA (SEQ ID NO: 4939), GHDSPHKSA (SEQ ID NO: 4940), GNYSPHKIG (SEQ ID NO: 4941), GHDSPHKSR (SEQ ID NO: 4942), GSGSPHSKL (SEQ ID NO: 4943), GSGSPHSRA (SEQ ID NO: 4944), GSGSPHSKR (SEQ ID NO: 4945), GSGSPHSLR (SEQ ID NO: 4946), GSGSPHSRG (SEQ ID NO: 4947), GSGSPHSSR (SEQ ID NO: 4948), RVGSPHSKA (SEQ ID NO: 4949), GSCSPHRKA (SEQ ID NO: 4950), GSGSPHFLR (SEQ ID NO: 4951), GSGSPHSKW (SEQ ID NO: 4952), GSGSPHSKS (SEQ ID NO: 4953), GLLSPHWKA (SEQ ID NO: 4954), GSGSPHVRR (SEQ ID NO: 4955), GSGSPHSKV (SEQ ID NO: 4956), MSGSPHSKA (SEQ ID NO: 4957), RNGSPHSKA (SEQ ID NO: 4958), TSGSPHSKA (SEQ ID NO: 4959), ISGSPHSKA (SEQ ID NO: 4960), GPGSPHSKA (SEQ ID NO: 4961), GSGSPHSKT (SEQ ID NO: 4962), ESGSPHSKA (SEQ ID NO: 4963), SSGSPHSKA (SEQ ID NO: 4964), GNGSPHSKA (SEQ ID NO: 4965), ASGSPHSKA (SEQ ID NO: 4966), NSGSPHSKA (SEQ ID NO: 4967), LSGSPHSKA (SEQ ID NO: 4968), GGGSPHSKA (SEQ ID NO: 4969), KSGSPHSKA (SEQ ID NO: 4970), GGGSPHSKS (SEQ ID NO: 4971), GSGSPHSKG (SEQ ID NO: 4972), HSGSPHSKA (SEQ ID NO: 4973), GTGSPHSKA (SEQ ID NO: 4974), PSGSPHSKA (SEQ ID NO: 4975), GSVSPHGKA (SEQ ID NO: 4976), RSGSPHSKA (SEQ ID NO: 4977), GSGSPHTKA (SEQ ID NO: 4978), GIGSPHSKA (SEQ ID NO: 4979), WSGSPHSKA (SEQ ID NO: 4980), DSGSPHSKA (SEQ ID NO: 4981), IDGSPHSKA (SEQ ID NO: 4982), GSGSPHNKA (SEQ ID NO: 4983), GLGSPHSKS (SEQ ID NO: 4984), DAGSPHSKA (SEQ ID NO: 4985), DGGSPHSKA (SEQ ID NO: 4986), MEGSPHSKA (SEQ ID NO: 4987), ENGSPHSKA (SEQ ID NO: 4988), GSASPHSKA (SEQ ID NO: 4989), GNGSPHSKS (SEQ ID NO: 4990), KNGSPHSKA (SEQ ID NO: 4991), KEGSPHSKA (SEQ ID NO: 4992), AIGSPHSKA (SEQ ID NO: 4993), GSGSPHSKN (SEQ ID NO: 4994), GSGSPHAKA (SEQ ID NO: 4995), GHDSPHKIG (SEQ ID NO: 4996), GYDSPHKSG (SEQ ID NO: 4997), GHESPHKSG (SEQ ID NO: 4998), GHDSPHKTG (SEQ ID NO: 4999), GRGSPHKRG (SEQ ID NO: 5000), GQDSPHKSG (SEQ ID NO: 4908), GHDSPHKSL (SEQ ID NO: 5001), GHGSPHSKA (SEQ ID NO: 5002), GHDSPHKSE (SEQ ID NO: 5003), VSGSPHSKA (SEQ ID NO: 4913), GRDSPHKSG (SEQ ID NO: 5004), GNDSPHKSV (SEQ ID NO: 5005), GQDSPHKIG (SEQ ID NO: 5006), GHDSPHKSV (SEQ ID NO: 5007), GPDSPHKIG (SEQ ID NO: 5008), GPDSPHKSG (SEQ ID NO: 5009), GHDSPHKSW (SEQ ID NO: 5010), GHDSPHKSN (SEQ ID NO: 5011), GMGSPHSKT (SEQ ID NO: 5012), GHDSPHKHG (SEQ ID NO: 5013), GQVSPHKSG (SEQ ID NO: 5014), GDDSPHKSV (SEQ ID NO: 5015), GHNSPHKSG (SEQ ID NO: 5016), GNGSPHKRG (SEQ ID NO: 5017), GHDSPHKYG (SEQ ID NO: 5018), GHDSPHKSQ (SEQ ID NO: 5019), GNDSPHKIG (SEQ ID NO: 5020), GHDSPHKSK (SEQ ID NO: 5021), GHDSPHKLW (SEQ ID NO: 5022), GHPSPHWKG (SEQ ID NO: 5023), GHDSPHKMG (SEQ ID NO: 5024), GHDSPHKMA (SEQ ID NO: 5025), or GHSSPHRSG (SEQ ID NO: 5026); an amino acid sequence comprising any portion of any of the aforesaid amino acid sequences (e.g., any 2, 3, 4, 5, 6, 7, or 8 amino acids, e.g., consecutive amino acids) thereof; an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the aforesaid amino acid sequences; or an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the aforesaid amino acid sequences. In some embodiments, [N1]-[N2]-[N3] is or comprises GSGSPHSKA (SEQ ID NO: 4697). In some embodiments, [N1]-[N2]-[N3] is or comprises GHDSPHKSG (SEQ ID NO: 4698).

In some embodiments, the AAV capsid variant comprising an amino acid sequence having the formula [N1]-[N2]-[N3], further comprises [N4], wherein [N4] comprises X7 X8 X9 X10. In some embodiments, position X7 of [N4] is W, Q, K, R, G, L, V, S, P, H, K, I, M, A, E, or F. In some embodiments, position X8 of [N4] is N, Y, C, K, T, H, R, D, V, S, P, G, W, E, F, A, I, M, Q, or L. In some embodiments, position X9 of [N4] is Q, G, K, H, R, T, L, D, A, P, I, F, V, M, W, Y, S, E, N, or Y. In some embodiments, position X10 of [N4] is Q, H, L, R, W, K, A, P, E, M, I, S, G, N, Y, C, V, T, D, or V. In some embodiments [N4] comprises QNQQ (SEQ ID NO: 5028), WNQQ (SEQ ID NO: 5029), QYYV (SEQ ID NO: 5030), RRQQ (SEQ ID NO: 5031), GCGQ (SEQ ID NO: 5032), LRQQ (SEQ ID NO: 5033), RNQQ (SEQ ID NO: 5034), VNQQ (SEQ ID NO: 5035), FRLQ (SEQ ID NO: 5036), FNQQ (SEQ ID NO: 5037), LLQQ (SEQ ID NO: 5038), SNQQ (SEQ ID NO: 5039), RLQQ (SEQ ID NO: 5040), LNQQ (SEQ ID NO: 5041), QRKL (SEQ ID NO: 5042), LRRQ (SEQ ID NO: 5043), QRLR (SEQ ID NO: 5044), QRRL (SEQ ID NO: 5045), RRLQ (SEQ ID NO: 5046), RLRQ (SEQ ID NO: 5047), SKRQ (SEQ ID NO: 5048), QLYR (SEQ ID NO: 5049), QLTV (SEQ ID NO: 5050), QNKQ (SEQ ID NO: 5051), KNQQ (SEQ ID NO: 5052), QKQQ (SEQ ID NO: 5053), QTQQ (SEQ ID NO: 5054), QNHQ (SEQ ID NO: 5055), QHQQ (SEQ ID NO: 5056), QNQH (SEQ ID NO: 5057), QHRQ (SEQ ID NO: 5058), LTQQ (SEQ ID NO: 5059), QNQW (SEQ ID NO: 5060), QNTH (SEQ ID NO: 5061), RRRQ (SEQ ID NO: 5062), QYQQ (SEQ ID NO: 5063), QNDQ (SEQ ID NO: 5064), QNRH (SEQ ID NO: 5065), RDQQ (SEQ ID NO: 5066), PNLQ (SEQ ID NO: 5067), HVRQ (SEQ ID NO: 5068), PNQH (SEQ ID NO: 5069), HNQQ (SEQ ID NO: 5070), QSQQ (SEQ ID NO: 5071), QPAK (SEQ ID NO: 5072), QNLA (SEQ ID NO: 5073), QNQL (SEQ ID NO: 5074), QGQQ (SEQ ID NO: 5075), LNRQ (SEQ ID NO: 5076), QNPP (SEQ ID NO: 5077), QNLQ (SEQ ID NO: 5078), QDQE (SEQ ID NO: 5079), QDQQ (SEQ ID NO: 5080), HWQQ (SEQ ID NO: 5081), PNQQ (SEQ ID NO: 5082), PEQQ (SEQ ID NO: 5083), QRTM (SEQ ID NO: 5084), LHQH (SEQ ID NO: 5085), QHRI (SEQ ID NO: 5086), QYIH (SEQ ID NO: 5087), QKFE (SEQ ID NO: 5088), QFPS (SEQ ID NO: 5089), QNPL (SEQ ID NO: 5090), QAIK (SEQ ID NO: 5091), QNRQ (SEQ ID NO: 5092), QYQH (SEQ ID NO: 5093), QNPQ (SEQ ID NO: 5094), QHQL (SEQ ID NO: 5095), QSPP (SEQ ID NO: 5096), QAKL (SEQ ID NO: 5097), KSQQ (SEQ ID NO: 5098), QDRP (SEQ ID NO: 5099), QNLG (SEQ ID NO: 5100), QAFH (SEQ ID NO: 5101), QNAQ (SEQ ID NO: 5102), HNQL (SEQ ID NO: 5103), QKLN (SEQ ID NO: 5104), QNVQ (SEQ ID NO: 5105), QAQQ (SEQ ID NO: 5106), QTPP (SEQ ID NO: 5107), QPPA (SEQ ID NO: 5108), QERP (SEQ ID NO: 5109), QDLQ (SEQ ID NO: 5110), QAMH (SEQ ID NO: 5111), QHPS (SEQ ID NO: 5112), PGLQ (SEQ ID NO: 5113), QGIR (SEQ ID NO: 5114), QAPA (SEQ ID NO: 5115), QIPP (SEQ ID NO: 5116), QTQL (SEQ ID NO: 5117), QAPS (SEQ ID NO: 5118), QNTY (SEQ ID NO: 5119), QDKQ (SEQ ID NO: 5120), QNHL (SEQ ID NO: 5121), QIGM (SEQ ID NO: 5122), LNKQ (SEQ ID NO: 5123), PNQL (SEQ ID NO: 5124), QLQQ (SEQ ID NO: 5125), QRMS (SEQ ID NO: 5126), QGIL (SEQ ID NO: 5127), QDRQ (SEQ ID NO: 5128), RDWQ (SEQ ID NO: 5129), QERS (SEQ ID NO: 5130), QNYQ (SEQ ID NO: 5131), QRTC (SEQ ID NO: 5132), QIGH (SEQ ID NO: 5133), QGAI (SEQ ID NO: 5134), QVPP (SEQ ID NO: 5135), QVQQ (SEQ ID NO: 5136), LMRQ (SEQ ID NO: 5137), QYSV (SEQ ID NO: 5138), QAIT (SEQ ID NO: 5139), QKTL (SEQ ID NO: 5140), QLHH (SEQ ID NO: 5141), QNII (SEQ ID NO: 5142), QGHH (SEQ ID NO: 5143), QSKV (SEQ ID NO: 5144), QLPS (SEQ ID NO: 5145), IGKQ (SEQ ID NO: 5146), QAIH (SEQ ID NO: 5147), QHGL (SEQ ID NO: 5148), QFMC (SEQ ID NO: 5149), QNQM (SEQ ID NO: 5150), QHLQ (SEQ ID NO: 5151), QPAR (SEQ ID NO: 5152), QSLQ (SEQ ID NO: 5153), QSQL (SEQ ID NO: 5154), HSQQ (SEQ ID NO: 5155), QMPS (SEQ ID NO: 5156), QGSL (SEQ ID NO: 5157), QVPA (SEQ ID NO: 5158), HYQQ (SEQ ID NO: 5159), QVPS (SEQ ID NO: 5160), RGEQ (SEQ ID NO: 5161), PGQQ (SEQ ID NO: 5162), LEQQ (SEQ ID NO: 5163), QNQS (SEQ ID NO: 5164), QKVI (SEQ ID NO: 5165), QNND (SEQ ID NO: 5166), QSVH (SEQ ID NO: 5167), QPLG (SEQ ID NO: 5168), HNQE (SEQ ID NO: 5169), QIQQ (SEQ ID NO: 5170), QVRN (SEQ ID NO: 5171), PSNQ (SEQ ID NO: 5172), QVGH (SEQ ID NO: 5173), QRDI (SEQ ID NO: 5174), QMPN (SEQ ID NO: 5175), RGLQ (SEQ ID NO: 5176), PSLQ (SEQ ID NO: 5177), QRDQ (SEQ ID NO: 5178), QAKG (SEQ ID NO: 5179), QSAH (SEQ ID NO: 5180), QSTM (SEQ ID NO: 5181), QREM (SEQ ID NO: 5182), QYRA (SEQ ID NO: 5183), QRQQ (SEQ ID NO: 5184), QWQQ (SEQ ID NO: 5185), QRMN (SEQ ID NO: 5186), GDSQ (SEQ ID NO: 5187), QKIS (SEQ ID NO: 5188), PSMQ (SEQ ID NO: 5189), SPRQ (SEQ ID NO: 5190), MEQQ (SEQ ID NO: 5191), QYQN (SEQ ID NO: 5192), QIRQ (SEQ ID NO: 5193), QSVQ (SEQ ID NO: 5194), RSQQ (SEQ ID NO: 5195), QNKL (SEQ ID NO: 5196), QIQH (SEQ ID NO: 5197), PRQQ (SEQ ID NO: 5198), HTQQ (SEQ ID NO: 5199), QRQH (SEQ ID NO: 5200), RNQE (SEQ ID NO: 5201), QSKQ (SEQ ID NO: 5202), QNQP (SEQ ID NO: 5203), QSPQ (SEQ ID NO: 5204), QTRQ (SEQ ID NO: 5205), QNLH (SEQ ID NO: 5206), QNQE (SEQ ID NO: 5207), LNQP (SEQ ID NO: 5208), QNQD (SEQ ID NO: 5209), QNLL (SEQ ID NO: 5210), QLVI (SEQ ID NO: 5211), RTQE (SEQ ID NO: 5212), QTHQ (SEQ ID NO: 5213), QDQH (SEQ ID NO: 5214), QSQH (SEQ ID NO: 5215), VRQQ (SEQ ID NO: 5216), AWQQ (SEQ ID NO: 5217), QSVP (SEQ ID NO: 5218), QNIQ (SEQ ID NO: 5219), LDQQ (SEQ ID NO: 5220), PDQQ (SEQ ID NO: 5221), ESQQ (SEQ ID NO: 5222), QRQL (SEQ ID NO: 5223), QIIV (SEQ ID NO: 5224), QKQS (SEQ ID NO: 5225), QSHQ (SEQ ID NO: 5226), QFVV (SEQ ID NO: 5227), QSQP (SEQ ID NO: 5228), QNEQ (SEQ ID NO: 5229), INQQ (SEQ ID NO: 5230), RNRQ (SEQ ID NO: 5231), RDQK (SEQ ID NO: 5232), QWKR (SEQ ID NO: 5233), ENRQ (SEQ ID NO: 5234), QTQP (SEQ ID NO: 5235), QKQL (SEQ ID NO: 5236), RNQL (SEQ ID NO: 5237), ISIQ (SEQ ID NO: 5238), QTVC (SEQ ID NO: 5239), QQIM (SEQ ID NO: 5240), LNHQ (SEQ ID NO: 5241), QNQA (SEQ ID NO: 5242), QMIH (SEQ ID NO: 5243), RNHQ (SEQ ID NO: 5244), or QKMN (SEQ ID NO: 5245), or any dipeptide or tripeptide thereof. In some embodiments, [N1]-[N2]-[N3]-[N4] is or comprises: the amino acid sequence of any of SEQ ID NOs: 1800-2241; an amino acid sequence comprising any portion of any of the aforesaid amino acid sequences (e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids, e.g., consecutive amino acids) thereof; an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the aforesaid amino acid sequences; or an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the aforesaid amino acid sequences. In some embodiments, [N1]-[N2]-[N3]-[N4] is or comprises GSGSPHSKAQNQQ (SEQ ID NO: 1801). In some embodiments, [N1]-[N2]-[N3]-[N4] is or comprises GHDSPHKSGQNQQ (SEQ ID NO: 1800).

In some embodiments, the AAV capsid variant comprising an amino acid sequence having the formula [N1]-[N2]-[N3], further comprises [N0], wherein [N0] comprises XA XB and XC. In some embodiments, XA of [N0] is T, S, Y, M, A, C, I, R, L, D, F, V, Q, N, H, E, or G. In some embodiments, XB of [N0] is I, M, P, E, N, D, S, A, T, G, Q, F, V, L, C, H, R, W, or L. In some embodiments, XC of [N0] is N, M, E, G, Y, W, T, I, Q, F, V, A, L, I, P, K, R, H, S, D, or S. In some embodiments, [N0] comprises TIN, SMN, TIM, YLS, GLS, MPE, MEG, MEY, AEW, CEW, ANN, IPE, ADM, IEY, ADY, IET, MEW, CEY, RIN, MEI, LEY, ADW, IEI, DIM, FEQ, MEF, CDQ, LPE, IEN, MES, AEI, VEY, IIN, TSN, IEV, MEM, AEV, MDA, VEW, AEQ, LEW, MEL, MET, MEA, IES, MEV, CEI, ATN, MDG, QEV, ADQ, NMN, IEM, ISN, TGN, QQQ, HDW, IEG, TII, TFP, TEK, EIN, TVN, TFN, SIN, TER, TSY, ELH, AIN, SVN, TDN, TFH, TVH, TEN, TSS, TID, TCN, NIN, TEH, AEM, AIK, TDK, TFK, SDQ, TEI, NTN, TET, SIK, TEL, TEA, TAN, TIY, TFS, TES, TTN, TED, TNN, EVH, TIS, TVR, TDR, TIK, NHI, TIP, ESD, TDL, TVP, TVI, AEH, NCL, TVK, NAD, TIT, NCV, TIR, NAL, VIN, TIQ, TEF, TRE, QGE, SEK, NVN, GGE, EFV, SDK, TEQ, EVQ, TEY, NCW, TDV, SDI, NSI, NSL, EVV, TEP, SEL, TWQ, TEV, AVN, GVL, TLN, TEG, TRD, NAI, AEN, AET, ETA, NNL, or any dipeptide thereof. In some embodiments, [N0]-[N1]-[N2]-[N3]-[N4] is or comprises the amino acid sequence of any one of SEQ ID NOs: 2242-2886; an amino acid sequence comprising any portion of any of the aforesaid amino acid sequences (e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids, e.g., consecutive amino acids) thereof; an amino acid sequence comprising one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the aforesaid amino acid sequences; or an amino acid sequence comprising one, two, or three but no more than four different amino acids, relative to any one of the aforesaid amino acid sequences. In some embodiments, [N0]-[N1]-[N2]-[N3]-[N4] is or comprises TINGSGSPHSKAQNQQ (SEQ ID NO: 2242). In some embodiments, [N0]-[N1]-[N2]-[N3]-[N4] is or comprises TINGHDSPHKSGQNQQ (SEQ ID NO: 2243).

In some embodiments, [N1]-[N2]-[N3] is present in loop IV of the AAV capsid variant. In some embodiments [N0] and [N4] are present in loop IV of the AAV capsid variant. In some embodiments, [N0]-[N1]-[N2]-[N3]-[N4] is present in loop IV of the AAV capsid variant. In some embodiments, [N0] is present immediately subsequent to position 449, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, wherein [N0] is present immediately subsequent to position 449, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981 or 982. In some embodiments, [N0] replaces positions 450, 451, and 452 (e.g., amino acids T450, 1451, and N452), relative to a reference sequence numbered according to SEQ ID NO: 138, 981, or 982. wherein [N0] is present immediately subsequent to position 449 and wherein [N0] replaces positions 450-452 (e.g., T450, I451, and N452), relative to a reference sequence numbered according to SEQ ID NO: 138, 981, or 982. In some embodiments, [N1] is present immediately subsequent to position 452, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138, 981 or 982. In some embodiments, wherein [N1] replaces positions 453-455 (e.g., G453, S454, and G455), relative to a reference sequence numbered according to SEQ ID NO: 138, 981, or 982. In some embodiments, [N1] is present immediately subsequent to position 452 and wherein [N1] replaces positions 453-455 (e.g., G453, S454, and G455), relative to a reference sequence numbered according to SEQ ID NO: 138, 981, or 982. In some embodiments, [N2] is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138, 981, or 982. In some embodiments, [N2]-[N3] is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138, 981, or 982. In some embodiments [N1]-[N2]-[N3] is present immediately subsequent to position 452, numbered relative to SEQ ID NO: 138, 981, or 982. In some embodiments, [N1]-[N2]-[N3] replaces positions 453-455 (e.g., G453, S454, and G455), relative to a reference sequence numbered according to SEQ ID NO: 138, 981, or 982. In some embodiments, [N1] is present immediately subsequent to position 452 and wherein [N1]-[N2]-[N3] replaces positions 453-455 (e.g., G453, S454, and G455), relative to a reference sequence numbered according to SEQ ID NO: 138, 981, or 982. In some embodiments, [N4] is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [N4] replaces positions 456-459 (e.g., Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [N4] is present immediately subsequent to position 455, and [N4] replaces positions 456-459 (e.g., Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [N2]-[N3]-[N4] replaces positions 456-459 (e.g., Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [N2]-[N3]-[N4] is present immediately subsequent to position 455, and wherein [N2]-[N3]-[N4] replaces positions 456-459 (e.g., Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [N1]-[N2]-[N3]-[N4] replaces positions 453-459 (e.g., G453, S454, G455, Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [N1]-[N2]-[N3]-[N4] is present immediately subsequent to position 452, and wherein [N1]-[N2]-[N3]-[N4] replaces positions 453-459 (e.g., G453, S454, G455, Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [N0]-[N1]-[N2]-[N3]-[N4] replaces positions 450-456 (e.g., T450, I451, N452, G453, 5454, G455, Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [N0]-[N1]-[N2]-[N3]-[N4] is present immediately subsequent to position 449, and wherein [N0]-[N1]-[N2]-[N3]-[N4] replaces positions 450-456 (e.g., T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, and Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.

In some embodiments, [N3] is present immediately subsequent to [N2].

In some embodiments, the AAV capsid variant comprises from N-terminus to C-terminus, [N2]-[N3]. In some embodiments, the AAV capsid variant comprises from N-terminus to C-terminus, [N1]-[N2]-[N3]. In some embodiments, the AAV capsid variant comprises from N-terminus to C-terminus, [N1]-[N2]-[N3]-[N4]. In some embodiments, the AAV capsid variant comprises from N-terminus to C-terminus, [N0]-[N1]-[N2]-[N3]. In some embodiments, the AAV capsid variant comprises from N-terminus to C-terminus, [N0]-[N1]-[N2]-[N3]-[N4].

In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence having the formula [A][B](SEQ ID NO: 4694), wherein [A] comprises the amino acid sequence of GSGSPH (SEQ ID NO: 4695) and [B] comprises X1 X2 X3 X4 X5 X6 X7. In some embodiments, position X1 of [B] is S, C, F, or V. In some embodiments, position X2 of [B] is K, L, R, I, E, Y, V, or S. In some embodiments, X3 of [B] is A, R, L, G, I, Y, S, F, or W. In some embodiments X4 of [B] is W, Q, R, G, L, V, S, or F. In some embodiments, position X5 of [B] is N, Y, R, C, K, or L. In some embodiments, position X6 of [B] is Q, G, K, R, T, L, or Y. In some embodiment, position X7 of [B] is Q, L, R, or V. In some embodiments, [B] comprises S LLWNQQ (SEQ ID NO: 5247), SKAQYYV (SEQ ID NO: 5248), SKLRRQQ (SEQ ID NO: 5249), SIWQNQQ (SEQ ID NO: 5250), SKAGCGQ (SEQ ID NO: 5251), SRAQNQQ (SEQ ID NO: 5252), SKRLRQQ (SEQ ID NO: 5253), SLRRNQQ (SEQ ID NO: 5254), SRGRNQQ (SEQ ID NO: 5255), SEIVNQQ (SEQ ID NO: 5256), SSRRNQQ (SEQ ID NO: 5257), CLLQNQQ (SEQ ID NO: 5258), SKAFRLQ (SEQ ID NO: 5259), CLAQNQQ (SEQ ID NO: 5260), FLRQNQQ (SEQ ID NO: 5261), SLRFNQQ (SEQ ID NO: 5262), SYLRNQQ (SEQ ID NO: 5263), CSLQNQQ (SEQ ID NO: 5264), VLWQNQQ (SEQ ID NO: 5265), SKWLLQQ (SEQ ID NO: 5266), SLWSNQQ (SEQ ID NO: 5267), SKRRLQQ (SEQ ID NO: 5268), SVYLNQQ (SEQ ID NO: 5269), SLWLNQQ (SEQ ID NO: 5270), SKAQRKL (SEQ ID NO: 5271), SKALRRQ (SEQ ID NO: 5272), SKAQRLR (SEQ ID NO: 5273), SKAQNQQ (SEQ ID NO: 5274), SKAQRRL (SEQ ID NO: 5275), SKARRQQ (SEQ ID NO: 5276), SKARRLQ (SEQ ID NO: 5277), SKSRRQQ (SEQ ID NO: 5278), SKARLRQ (SEQ ID NO: 5279), SKASKRQ (SEQ ID NO: 5280), VRRQNQQ (SEQ ID NO: 5281), SKAQLYR (SEQ ID NO: 5282), SLFRNQQ (SEQ ID NO: 5283), SKAQLTV (SEQ ID NO: 5284), or any dipeptide, tripeptide, tetrapeptide, pentapeptide, or hexapeptide thereof. In some embodiments, [A][B] comprises GSGSPHSLLWNQQ (SEQ ID NO: 5285), GSGSPHSKAQYYV (SEQ ID NO: 2060), GSGSPHSKLRRQQ (SEQ ID NO: 2061), GSGSPHSIWQNQQ (SEQ ID NO: 5286), GSGSPHSKAGCGQ (SEQ ID NO: 2062), GSGSPHSRAQNQQ (SEQ ID NO: 2063), GSGSPHSKRLRQQ (SEQ ID NO: 2064), GSGSPHSLRRNQQ (SEQ ID NO: 2065), GSGSPHSRGRNQQ (SEQ ID NO: 2066), GSGSPHSEIVNQQ (SEQ ID NO: 5287), GSGSPHSSRRNQQ (SEQ ID NO: 2067), GSGSPHCLLQNQQ (SEQ ID NO: 5288), GSGSPHSKAFRLQ (SEQ ID NO: 2068), GSGSPHCLAQNQQ (SEQ ID NO: 5289), GSGSPHFLRQNQQ (SEQ ID NO: 2070), GSGSPHSLRFNQQ (SEQ ID NO: 2071), GSGSPHSYLRNQQ (SEQ ID NO: 5290), GSGSPHCSLQNQQ (SEQ ID NO: 5291), GSGSPHVLWQNQQ (SEQ ID NO: 5292), GSGSPHSKWLLQQ (SEQ ID NO: 2072), GSGSPHSLWSNQQ (SEQ ID NO: 5293), GSGSPHSKRRLQQ (SEQ ID NO: 2073), GSGSPHSVYLNQQ (SEQ ID NO: 5294), GSGSPHSLWLNQQ (SEQ ID NO: 5295), GSGSPHSKAQRKL (SEQ ID NO: 2074), GSGSPHSKALRRQ (SEQ ID NO: 2075), GSGSPHSKAQRLR (SEQ ID NO: 2076), GSGSPHSKAQNQQ (SEQ ID NO: 1801), GSGSPHSKAQRRL (SEQ ID NO: 2077), GSGSPHSKARRQQ (SEQ ID NO: 2078), GSGSPHSKARRLQ (SEQ ID NO: 2079), GSGSPHSKSRRQQ (SEQ ID NO: 2080), GSGSPHSKARLRQ (SEQ ID NO: 2082), GSGSPHSKASKRQ (SEQ ID NO: 2083), GSGSPHVRRQNQQ (SEQ ID NO: 2084), GSGSPHSKAQLYR (SEQ ID NO: 2085), GSGSPHSLFRNQQ (SEQ ID NO: 5296), GSGSPHSKAQLTV (SEQ ID NO: 2086), or any portion thereof, e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids, e.g., consecutive amino acids, thereof.

In some embodiments, [A][B] is present in loop IV of the AAV capsid variant. In some embodiments, [A] is present immediately subsequent to position 452, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [A] replaces positions 453-455 (e.g., G453, S454, G455), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [A] is present immediately subsequent to position 452, and wherein [A] replaces positions 453-455 (e.g., G453, S454, G455), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [B] is present immediately subsequent to [A]. In some embodiments, [B] replaces positions 456-459 (e.g., Q456, N457, Q458, Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [A][B] replaces positions 453-459 (e.g., G453, S454, G455, Q456, N457, Q458, Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [A][B] is present immediately subsequent to position 452, and wherein [A][B] replaces positions 453-459 (e.g., G453, S454, G455, Q456, N457, Q458, Q459), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises from N-terminus to C-terminus, [A][B].

In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence having the formula [A][B](SEQ ID NO: 4699), wherein [A] comprises X1 X2 X3 X4 X5 X6 and [B] comprises SPHKSG (SEQ ID NO: 946). In some embodiments, position X1 of [A] is T, M, A, C, I, R, L, D, F, V, Q, N, or H. In some embodiments, position X2 of [A] is I, P, E, N, D, S, A, T, M, or Q. In some embodiments, position X3 of [A] is N, E, G, Y, W, M, T, I, K, Q, F, S, V, A, or L. In some embodiments, position X4 of [A] is G, D, R, or E. In some embodiments, position X5 of [A] is H, Q, N, or D. In some embodiments, position X6 of [A] is D or R. In some embodiments, [A] comprises TINGHD (SEQ ID NO: 5297), MPEGHD (SEQ ID NO: 5298), MEGGHD (SEQ ID NO: 5299), MEYGHD (SEQ ID NO: 5300), AEWGHD (SEQ ID NO: 5301), CEWGHD (SEQ ID NO: 5302), ANNGQD (SEQ ID NO: 5303), IPEGHD (SEQ ID NO: 5304), ADMGHD (SEQ ID NO: 5305), IEYGHD (SEQ ID NO: 5306), ADYGHD (SEQ ID NO: 5307), IETGHD (SEQ ID NO: 5308), MEWGHD (SEQ ID NO: 5309), CEYGHD (SEQ ID NO: 5310), RINGHD (SEQ ID NO: 5311), MEIGHD (SEQ ID NO: 5312), LEYGHD (SEQ ID NO: 5313), ADWGHD (SEQ ID NO: 5314), IEIGHD (SEQ ID NO: 5315), TIKDND (SEQ ID NO: 5316), DIMGHD (SEQ ID NO: 5317), FEQGHD (SEQ ID NO: 5318), MEFGHD (SEQ ID NO: 5319), CDQGHD (SEQ ID NO: 5320), LPEGHD (SEQ ID NO: 5321), IENGHD (SEQ ID NO: 5322), MESGHD (SEQ ID NO: 5323), AEIGHD (SEQ ID NO: 5324), VEYGHD (SEQ ID NO: 5325), TSNGDD (SEQ ID NO: 5326), IEVGHD (SEQ ID NO: 5327), MEMGHD (SEQ ID NO: 5328), AEVGHD (SEQ ID NO: 5329), MDAGHD (SEQ ID NO: 5330), VEWGHD (SEQ ID NO: 5331), AEQGHD (SEQ ID NO: 5332), LEWGHD (SEQ ID NO: 5333), MELGHD (SEQ ID NO: 5334), METGHD (SEQ ID NO: 5335), MEAGHD (SEQ ID NO: 5336), TINRQR (SEQ ID NO: 5337), IESGHD (SEQ ID NO: 5338), TAKDHD (SEQ ID NO: 5339), MEVGHD (SEQ ID NO: 5340), CEIGHD (SEQ ID NO: 5341), ATNGHD (SEQ ID NO: 5342), MDGGHD (SEQ ID NO: 5343), QEVGHD (SEQ ID NO: 5344), ADQGHD (SEQ ID NO: 5345), NMNGHD (SEQ ID NO: 5346), TPWEHD (SEQ ID NO: 5347), IEMGHD (SEQ ID NO: 5348), TANEHD (SEQ ID NO: 5349), QQQGHD (SEQ ID NO: 5350), TPQDHD (SEQ ID NO: 5351), HDWGHD (SEQ ID NO: 5352), IEGGHD (SEQ ID NO: 5353), or any dipeptide, tripeptide, tetrapeptide, or pentapeptide thereof. In some embodiments, [A][B] comprises TINGHDSPHKR (SEQ ID NO: 5354), MPEGHDSPHKS (SEQ ID NO: 5355), MEGGHDSPHKS (SEQ ID NO: 5356), MEYGHDSPHKS (SEQ ID NO: 5357), AEWGHDSPHKS (SEQ ID NO: 5358), CEWGHDSPHKS (SEQ ID NO: 5359), ANNGQDSPHKS (SEQ ID NO: 5360), IPEGHDSPHKS (SEQ ID NO: 5361), ADMGHDSPHKS (SEQ ID NO: 5362), IEYGHDSPHKS (SEQ ID NO: 5363), ADYGHDSPHKS (SEQ ID NO: 5364), IETGHDSPHKS (SEQ ID NO: 5365), MEWGHDSPHKS (SEQ ID NO: 5366), CEYGHDSPHKS (SEQ ID NO: 5367), RINGHDSPHKS (SEQ ID NO: 5368), MEIGHDSPHKS (SEQ ID NO: 5369), LEYGHDSPHKS (SEQ ID NO: 5370), ADWGHDSPHKS (SEQ ID NO: 5371), IEIGHDSPHKS (SEQ ID NO: 5372), TIKDNDSPHKS (SEQ ID NO: 5373), DIMGHDSPHKS (SEQ ID NO: 5374), FEQGHDSPHKS (SEQ ID NO: 5375), MEFGHDSPHKS (SEQ ID NO: 5376), CDQGHDSPHKS (SEQ ID NO: 5377), LPEGHDSPHKS (SEQ ID NO: 5378), IENGHDSPHKS (SEQ ID NO: 5379), MESGHDSPHKS (SEQ ID NO: 5380), AEIGHDSPHKS (SEQ ID NO: 5381), VEYGHDSPHKS (SEQ ID NO: 5382), TSNGDDSPHKS (SEQ ID NO: 5383), IEVGHDSPHKS (SEQ ID NO: 5384), MEMGHDSPHKS (SEQ ID NO: 5385), AEVGHDSPHKS (SEQ ID NO: 5386), MDAGHDSPHKS (SEQ ID NO: 5387), VEWGHDSPHKS (SEQ ID NO: 5388), AEQGHDSPHKS (SEQ ID NO: 5389), LEWGHDSPHKS (SEQ ID NO: 5390), MELGHDSPHKS (SEQ ID NO: 5391), METGHDSPHKS (SEQ ID NO: 5392), MEAGHDSPHKS (SEQ ID NO: 5393), TINRQRSPHKS (SEQ ID NO: 5394), IESGHDSPHKS (SEQ ID NO: 5395), TAKDHDSPHKS (SEQ ID NO: 5396), MEVGHDSPHKS (SEQ ID NO: 5397), CEIGHDSPHKS (SEQ ID NO: 5398), ATNGHDSPHKS (SEQ ID NO: 5399), MDGGHDSPHKS (SEQ ID NO: 5400), QEVGHDSPHKS (SEQ ID NO: 5401), ADQGHDSPHKS (SEQ ID NO: 5402), NMNGHDSPHKS (SEQ ID NO: 5403), TPWEHDSPHKS (SEQ ID NO: 5404), IEMGHDSPHKS (SEQ ID NO: 5405), TANEHDSPHKS (SEQ ID NO: 5406), TINGHDSPHKS (SEQ ID NO: 5407), QQQGHDSPHKS (SEQ ID NO: 5408), TPQDHDSPHKS (SEQ ID NO: 5409), HDWGHDSPHKS (SEQ ID NO: 5410), IEGGHDSPHKS (SEQ ID NO: 5411), or any portion thereof, e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids, e.g., consecutive amino acids, thereof.

In some embodiments, [A][B] is present in loop IV of the AAV capsid variant. In some embodiments, [A] is present immediately subsequent to position 449, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [A] replaces positions 450-455 (e.g., T450, 1451, N452, G453, S454, G455), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [A] is present immediately subsequent to position 449, and wherein [A] replaces positions 450-455 (e.g., T450, 1451, N452, G453, S454, G455), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [B] is present immediately subsequent to [A]. In some embodiments, [A][B] replaces positions 450-455 (e.g., T450, 1451, N452, G453, S454, G455), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, [A][B] is present immediately subsequent to position 449, and wherein [A][B] replaces positions 450-455 (e.g., T450, 1451, N452, G453, S454, G455), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the peptide comprises from N-terminus to C-terminus, [A][B].

In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 16, or 17 consecutive amino acids from any one of the sequences provided in Tables 1, 2A, 2B, 13-19. In some embodiments, the AAV capsid variant comprises an amino acid sequence comprising at least 3, 4, or 5 consecutive amino acids from any one of SEQ ID NOs: 945-980 or 985-986. In some embodiments, the AAV capsid variant comprises an amino acid sequence comprising at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 consecutive amino acids from any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909. In some embodiments, the amino acid sequence is present in loop IV. In some embodiments, the amino acid sequence is present immediately subsequent to position 448, 452, 453, 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138, 981, or 982. In some embodiments, the amino acid sequence is present immediately subsequent to position 455, numbered according to SEQ ID NO: 982. In some embodiments, the amino acid sequence is present immediately subsequent to position 455, numbered according to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 453, numbered according to SEQ ID NO: 981. In some embodiments, the amino acid sequence is present immediately subsequent to position 453, numbered according to SEQ ID NO: 138. In some embodiments, the amino acid sequence replaces 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or all of positions 499 (e.g., K499), 450 (e.g., T450), 451 (e.g., I451), 452 (e.g., N452), 453 (e.g., G453), 454 (e.g., S454), 455 (e.g., G455), 456 (e.g., Q456), 457 (e.g., N457), 458 (e.g., Q458), 459 (e.g., Q459), and 460 (e.g., T460), numbered according to SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises one or more amino acid substitutions at positions 499 (e.g., K499), 450 (e.g., T450), 451 (e.g., I451), 452 (e.g., N452), 453 (e.g., G453), 454 (e.g., S454), 455 (e.g., G455), 456 (e.g., Q456), 457 (e.g., N457), 458 (e.g., Q458), 459 (e.g., Q459), and/or 460 (e.g., T460), numbered according to SEQ ID NO: 138.

In some embodiments, the 3 consecutive amino acids comprise SPH. In some embodiments, the 4 consecutive amino acids comprise SPHS (SEQ ID NO: 4700). In some embodiments, the 5 consecutive amino acids comprise SPHSK (SEQ ID NO: 4701). In some embodiments, the 6 consecutive amino acids comprise SPHSKA (SEQ ID NO: 941).

In some embodiments, 3 consecutive amino acids comprise HDS. In some embodiments, the 4 consecutive amino acids comprise HDSP (SEQ ID NO: 4702). In some embodiments, the 5 consecutive amino acids comprise HDSPH (SEQ ID NO: 4703). In some embodiments, the 6 consecutive amino acids comprise HDSPHK (SEQ ID NO: 2).

In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of any one of the sequences provided in Tables 1, 2A, 2B, 13-19. In some embodiments, the AAV capsid variant comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids, relative to the amino acid sequence of any one of the sequences provided in Tables 1, 2A, 2B, 13-19. In some embodiments, the AAV capsid variant comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of any one of SEQ ID NOs: 945-980 or 985-986. In some embodiments, the AAV capsid variant comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids, relative to the amino acid sequence of any one of SEQ ID NOs: 945-980 or 985-986. In some embodiments, the AAV capsid variant comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909. In some embodiments, the AAV capsid variant comprises an amino acid sequence comprising at least one, two, or three but no more than four different amino acids, from the amino acid sequence of any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909. In some embodiments, the amino acid sequence is present in loop IV. In some embodiments, the amino acid sequence is present immediately subsequent to position 448, 452, 453, 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138, 981, or 982. In some embodiments, the amino acid sequence is present immediately subsequent to position 455, numbered according to SEQ ID NO: 982. In some embodiments, the amino acid sequence is present immediately subsequent to position 455, numbered according to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 453, numbered according to SEQ ID NO: 981. In some embodiments, the amino acid sequence is present immediately subsequent to position 453, numbered according to SEQ ID NO: 138. In some embodiments, the amino acid sequence replaces 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or all of positions 499 (e.g., K499), 450 (e.g., T450), 451 (e.g., I451), 452 (e.g., N452), 453 (e.g., G453), 454 (e.g., S454), 455 (e.g., G455), 456 (e.g., Q456), 457 (e.g., N457), 458 (e.g., Q458), 459 (e.g., Q459), and 460 (e.g., T460), numbered according to SEQ ID NO: 138.

In some embodiments, the AAV capsid variant comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941). In some embodiments, the AAV capsid variant comprises an amino acid sequence comprising at least one, two, or three, but no more than four different amino acids from the amino acid sequence of SPHSKA (SEQ ID NO: 941).

In some embodiments, the AAV capsid variant comprises an amino acid sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2). In some embodiments, the AAV capsid variant comprises an amino acid sequence comprising at least one, two, or three, but no more than four different amino acids that relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2).

In some embodiments, the AAV capsid variant, comprises the amino acid sequence of any of the sequences provided in Tables 1, 2A, 2B, 13-19. In some embodiments, the peptide comprises the amino acid sequence of any of SEQ ID NOs: 945-980 or 985-986. In some embodiments, the AAV capsid variant comprises the amino acid sequence of any of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909. In some embodiments, the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 941. In some embodiments, the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 2. In some embodiments, the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 943. In some embodiments, the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 3589. In some embodiments, the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 1754. In some embodiments, the amino acid sequence is present in loop IV. In some embodiments, the amino acid sequence is present immediately subsequent to position 448, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the amino acid sequence replaces positions 449-460 (e.g., K449, T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 448 and replaces positions 449-460 (e.g., K449, T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 449, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the amino acid sequence replaces positions 450-460 (e.g., T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 449, and replaces positions 450-460 (e.g., T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 450, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the amino acid sequence replaces positions 451-460 (e.g., I451, N452, G453, S454, G455, Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 450 and replaces positions 451-460 (e.g., I451, N452, G453, S454, G455, Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 451, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the amino acid sequence replaces positions 452-460 (e.g., N452, G453, S454, G455, Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 451 and replaces positions 452-460 (e.g., N452, G453, S454, G455, Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 452, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the amino acid sequence replaces positions 453-460 (e.g., G453, S454, G455, Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 452, and replaces positions 453-460 (e.g., G453, S454, G455, Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 453, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the amino acid sequence replaces positions 454 and 455 (e.g., S454 and G455), numbered according to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 453, and replaces positions 454 and 455 (e.g., S454 and G455), numbered according to SEQ ID NO: 138. In some embodiments, the amino acid sequence replaces positions 454-460 (e.g., S454, G455, Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 453, and replaces positions 454-460 (e.g., S454, G455, Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 454, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 454, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981. In some embodiments, the amino acid sequence replaces positions 455-460 (e.g., positions G455, Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to positions 454, and replaces positions 455-460 (e.g., positions G455, Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 982. In some embodiments, the amino acid sequence replaces positions 456-460 (e.g., Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138. In some embodiments, the amino acid sequence is present immediately subsequent to position 455, and replaces positions 456-460 (e.g., Q456, N457, Q458, Q459, and T460), numbered relative to SEQ ID NO: 138.

In some embodiments, the AAV capsid variant (e.g., an AAV capsid variant described herein), comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 942 or 944, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto. In some embodiments, the AAV capsid variant described herein, comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 3 or 942, or a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 3 or 942. In some embodiments, the AAV capsid variant comprises an amino acid sequence encoded by a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides relative to the nucleotide sequence of SEQ ID NO: 3 or 942.

In some embodiments, the nucleotide sequence encoding the AAV capsid variant (e.g., an AAV capsid variant described herein), comprises the nucleotide sequence of SEQ ID NO: 942, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto. In some embodiments, the nucleic acid sequence encoding the AAV capsid variant comprises a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequences of SEQ ID NO: 942. In some embodiments, the nucleotide sequence encoding an AAV capsid variant described herein comprises a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides, relative to the nucleotide sequence of SEQ ID NO: 942.

In some embodiments, the nucleotide sequence encoding the AAV capsid variant (e.g., an AAV capsid variant described herein), comprises the nucleotide sequence of SEQ ID NO: 3, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto. In some embodiments, the nucleic acid sequence encoding the AAV capsid variant comprises a nucleotide sequence comprising at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequences of SEQ ID NO: 3. In some embodiments, the nucleotide sequence encoding an AAV capsid variant described herein comprises a nucleotide sequence comprising at least one, two, three, four, five, six, or seven, but no more than ten different nucleotides relative to the nucleotide sequence of SEQ ID NO: 3.

In some embodiments, an AAV capsid variant described herein comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, an AAV capsid variant described herein comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 981.

In some embodiments, an AAV capsid variant described herein comprises the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein the amino acid sequence is present immediately subsequent to position 453, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, an AAV capsid variant described herein comprises the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein the amino acid sequence is present immediately subsequent to position 453, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 982.

In some embodiments, an AAV capsid variant described herein comprises (i) the amino acid sequence of HDSPHSKA (SEQ ID NO: 4486), which is present immediately subsequent to position 453; and (ii) a deletion of amino acids SG at position 454 and 455; wherein (i) and (ii) are numbered according to SEQ ID NO: 138.

In some embodiments, an AAV capsid variant described herein comprises an amino acid other than S at position 454 and/or an amino acid other than G at position 455, numbered according to SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises the amino acid H at position 454 and the amino acid D at position 455, numbered according to SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941). In some embodiments, the AAV capsid variant comprises: (i) the amino acid H at position 454 and the amino acid D at position 455, and (ii) the amino acid sequence SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence of SPHKSG (SEQ ID NO: 946) is present immediately subsequent to position 455, wherein (i) and (ii) are numbered according to SEQ ID NO: 138.

In some embodiments, an AAV capsid variant described herein comprises a modification, e.g., substitution, relative to SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises a modification, e.g., substitution, at position S454 and/or G455, numbered relative to SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises a S454H substitution and/or G455D substitution, numbered relative to SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises a S454H substitution and a G455D substitution, numbered relative to SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941). In some embodiments, the AAV capsid variant comprises: (i) a S454H substitution and a G455D substitution, and (ii) the amino acid sequence SPHKSG (SEQ ID NO: 946), wherein the amino acid sequence of SPHSKA (SEQ ID NO: 941) is present immediately subsequent to position 455, wherein (i) and (ii) are numbered according to SEQ ID NO: 138.

In some embodiments, the AAV capsid variant further comprises one, two, or all of an amino acid other than T at position 450 (e.g., S, Y, or G), an amino acid other than I at position 451 (e.g., M or L), and/or an amino acid other than N at position 452 (e.g., S), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises an S at position 450 and an M at position 451, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises a Y at position 450, an L at position 451, and an S at position 452, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises a G at position 450, an L at position 451, and an S at position 452, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.

In some embodiments, the AAV capsid variant further comprises one, two, three, four, or all of an amino acid other than Q at position 456 (e.g., R or L), N at position 457 (e.g., H, K, or R), Q at position 458 (e.g., R or T), Q at position 459 (H), and/or T at position 460 (N or S), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises an R at position 456, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises an L at position 456, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises an H at position 457 and an R at position 458, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises a K at position 457 and an N at position 460, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises a T at position 458, an H at position 459, and an S at position 460, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises an R at position 456, an R at position 457, and an R at position 458, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138.

In some embodiments, an AAV capsid variant described herein comprises an amino acid other than I at position 451, an amino acid other than N at position 452, and an amino acid other than G at position 453, numbered according to SEQ ID NO: 138 or 981. In some embodiments, the AAV capsid variant comprises E at position 451, R at position 452, and V at position 453, numbered according to SEQ ID NO: 138 or 981. In some embodiments, the AAV capsid variant comprises the substitutions 1451E, N452R, and G453V, numbered according to SEQ ID NO: 138 or 981.

In some embodiments, the AAV capsid variant comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to position 455 and wherein the AAV capsid variant comprises the E at position 451, R at position 452, and V at position 453, numbered according to the amino acid sequence of SEQ ID NO: 138 or 981. In some embodiments, the AAV capsid variant comprises the substitutions I451E, N452R, and G453V, and further comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to position 455, all numbered according to SEQ ID NO: 138 or 981. In some embodiments, the AAV capsid variant comprises the amino acid sequence of ERVSGSPHSKA (SEQ ID NO: 6399), and wherein the amino acid sequence is present immediately subsequent to position 449 and replaces positions 450-455, numbered according to SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589), wherein the amino acid sequence is present immediately subsequent to position 448 and replaces positions 449-460, numbered according to SEQ ID NO: 138.

In some embodiments, an AAV capsid variant described herein comprises an amino acid other than T at position 450, an amino acid other than I at position 451, and an amino acid other than N at position 452, numbered according to SEQ ID NO: 138 or 982. In some embodiments, the AAV capsid variant comprises A at position 450, E at position 451, and I at position 452, numbered according to SEQ ID NO: 138 or 982. In some embodiments, the AAV capsid variant comprises the substitutions T450A, 1451E, and N452I, numbered according to SEQ ID NO: 138 or 982.

In some embodiments, the AAV capsid variant comprises the amino acid sequence of HDSPHK (SEQ ID NO: 2), which is present immediately subsequent to positions 453, and further comprises A at position 450, E at position 451, and I at position 452, all numbered according to SEQ ID NO: 138 or 982. In some embodiments, the AAV capsid variant comprises the substitutions T450A, I451E, and N452I, and further comprises the amino acid sequence HDSPHK (SEQ ID NO: 2) present immediately subsequent to position 453, all numbered according to SEQ ID NO: 138 or 982. In some embodiments, the AAV capsid variant comprises the amino acid sequence of AEIGHDSPHKSG (SEQ ID NO: 6400), wherein the amino acid sequence is present immediately subsequent to position 449 and replaces positions 450-455, numbered according to SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises the amino acid sequence of KAEIGHDSPHKSGQNQQT (SEQ ID NO: 1754), wherein the amino acid sequence is present immediately subsequent to position 448 and replaces positions 449-460, numbered according to SEQ ID NO: 138.

In some embodiments, the AAV capsid variant, further comprises a substitution at position K449, e.g., a K449R substitution, numbered according to SEQ ID NO: 138. In some embodiments, the AAV capsid variant, further comprises an amino acid other than K at position 449 (e.g., R), relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises an R at position 449, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises a modification, e.g., an insertion, substitution, and/or deletion in loop I, II, VI, and/or VIII.

In some embodiments, the AAV capsid variant, further comprises an amino acid sequence comprising at least one, two or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, of the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant, further comprises an amino acid sequence comprising at least one, two or three, but not more than 30, 20 or 10 amino acids that differ from the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises the amino acid sequence of SEQ ID NO: 138, or an amino acid sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.

In some embodiments, the AAV capsid variant further comprises (a) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 138, 981, or 982; (b) a VP2 protein comprising the amino acid sequence of positions 138-736 of SEQ ID NO: 138 or positions 138-742 of SEQ ID NO: 981 or 982; (c) a VP3 protein comprising the amino acid sequence of positions 203-736 of SEQ ID NO: 138 or positions 203-742 of SEQ ID NO: 981 or 982; or (d) an amino acid sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity to any of the amino acid sequences in (a)-(c), an amino acid sequence comprising at least one, two or three, but not more than 30, 20 or 10 different amino acids relative to any of the amino acid sequences in (a)-(c), or an amino acid sequence comprising at least one, two or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to any of the amino acid sequences in (a)-(c).

In some embodiments, the AAV capsid variant further comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 137, or a sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, the AAV capsid variant further comprises an amino acid sequence encoded by a nucleotide sequence comprising at least one, two or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 137. In some embodiments, the AAV capsid variant further comprises an amino acid sequence encoded by a nucleotide sequence comprising at least one, two or three, but not more than 30, 20 or 10 different nucleotides, relative to the amino acid sequence of SEQ ID NO: 137.

In some embodiments, the nucleotide sequence encoding the AAV capsid variant further comprises the nucleotide sequence of SEQ ID NO: 137, or a sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, the nucleotide sequence encoding the AAV capsid variant further comprises a nucleotide sequence comprising at least one, two or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 137. In some embodiments, the nucleotide sequence encoding the AAV capsid variant further comprises a nucleotide sequence comprising at least one, two or three, but not more than 30, 20 or 10 different nucleotides, relative to the amino acid sequence of SEQ ID NO: 137.

In some embodiments, an AAV capsid variant of the present disclosure comprises an amino acid sequence as described herein, e.g., an amino acid sequence of an AAV capsid variant of TTM-001 or TTM-002, e.g., as described in Tables 3 and 4. In some embodiments, an AAV capsid variant of the present disclosure comprises an amino acid sequence as described herein, e.g., an amino acid sequence of an AAV capsid variant of TTM-003, TTM-004, TTM-005, TTM-006, TTM-007, TTM-008, TTM-009, TTM-010, TTM-011, TTM-012, TTM-013, TTM-014, TTM-015, TTM-016, TTM-017, TTM-018, TTM-019, TTM-020, TTM-021, TTM-022, TTM-023, TTM-024, TTM-025, or TTM-026, e.g., as described in Table 4.

In some embodiments, an AAV capsid variant described herein comprises a VP1, VP2, and/or VP3 protein comprising an amino acid sequence described herein, e.g., an amino acid sequence of an AAV capsid variant of TTM-001 or TTM-002, e.g., as described in Tables 3 and 4. In some embodiments, an AAV capsid variant described herein comprises a VP1, VP2, and/or VP3 protein comprising an amino acid sequence described herein, e.g., an amino acid sequence of an AAV capsid variant of TTM-003, TTM-004, TTM-005, TTM-006, TTM-007, TTM-008, TTM-009, TTM-010, TTM-011, TTM-012, TTM-013, TTM-014, TTM-015, TTM-016, TTM-017, TTM-018, TTM-019, TTM-020, TTM-021, TTM-022, TTM-023, TTM-024, TTM-025, or TTM-026, e.g., as described in Table 4.

In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence encoded by a nucleotide sequence as described herein, e.g., a nucleotide sequence of an AAV capsid variant of TTM-001 or TTM-002, e.g., as described in Tables 3 and 5. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence encoded by a nucleotide sequence as described herein, e.g., a nucleotide sequence of an AAV capsid variant of TTM-003, TTM-004, TTM-005, TTM-006, TTM-007, TTM-008, TTM-009, TTM-010, TTM-011, TTM-012, TTM-013, TTM-014, TTM-015, TTM-016, TTM-017, TTM-018, TTM-019, TTM-020, TTM-021, TTM-022, TTM-023, TTM-024, TTM-025, or TTM-026, e.g., as described in Table 5.

In some embodiments, a polynucleotide or nucleic acid encoding an AAV capsid variant, of the present disclosure comprises a nucleotide sequence described herein, e.g., a nucleotide sequence of an AAV capsid variant of TTM-001 or TTM-002, e.g., as described in Tables 3 and 5. In some embodiments, a polynucleotide or nucleic acid encoding an AAV capsid variant, of the present disclosure comprises a nucleotide sequence described herein, e.g., a nucleotide sequence of an AAV capsid variant of TTM-003, TTM-004, TTM-005, TTM-006, TTM-007, TTM-008, TTM-009, TTM-010, TTM-011, TTM-012, TTM-013, TTM-014, TTM-015, TTM-016, TTM-017, TTM-018, TTM-019, TTM-020, TTM-021, TTM-022, TTM-023, TTM-024, TTM-025, or TTM-026, e.g., as described in Table 5.

TABLE 3

Exemplary full length capsid sequences

VP1
Peptide
Peptide

VP1 DNA
(amino acid)
(amino acid)
DNA SEQ

Name
SEQ ID NO:
SEQ ID NO:
SEQ ID NO:
ID NO:

TTM-001
983
981
941
942

TTM-002
984
982
2
3

TABLE 4

Exemplary full length capsid amino acid sequences

SEQ

Name and
ID

Annotation
NO:
Amino Acid Sequence

TTM-001
981
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVEMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

742 aa

KTINGSGSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-002
982
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 454

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVEMIPQYGYLTLNDGSQAVGRS

position 453);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

742 aa

KTINGHDSPHKSGQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-003
36
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVEMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTERVSGSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 451,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

452, and 453

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-004
37
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 454

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 453);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KAEIGHDSPHKSGQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 450,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

451, and 452

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSLITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-005
38
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTIIGSGSPHSKAQNRHTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 452,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

464, and 465

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-006
39
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTEKMSGSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 451,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

452, and 453

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-007
40
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KEINGRGSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 450

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

and 454

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-008
41
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVEMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTFNGSGSPHSKAPNLQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 451,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

462, and 464

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-009
42
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTEKTSGSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 451,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

452, and 453

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-010
43
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTMNGHDSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 451,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

454, and 455

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-011
44
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTIDGHDSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 452,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

454, and 455

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-012
45
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTNNGHDSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 452,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

454, and 455

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-013
46
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVEMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTQRKSGSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 451,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

452, and 453

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-014
47
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVEMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTINGSGSPHSKAQARKTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 463,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

464, and 465

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-015
48
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KYIVGSGSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 450

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

and 452

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAENKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-016
49
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTISKRGSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 452,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

453, and 454

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-017
50
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KGLGGSGSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 450,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

451, and 452

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAENKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-018
51
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTINGHDSPHSKAQNLQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 454,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

455, and 464

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-019
52
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTVNGHDSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 451,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

454, and 455

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAENKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-020
53
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTINGHDSPHSKALNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 454,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

455, and 462

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAENKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-021
54
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTINGHDSPHSKAQNQQSLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 454,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

455, and 466

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-022
55
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTINGHDSPHSKAQNQQILKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 454,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

455, and 466

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAENKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-023
56
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVEMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

742 aa

KTINGSGSPHFTRQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-024
57
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modifications at

KTSNGHDSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 451,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

454, and 455

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-025
58
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modification at

KTINGSGSPHSLPWNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

position 462

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

underlined;

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

742 aa

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TTM-026
59
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPG

6mer peptide

NGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGG

underlined,

NLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAK

starts at

KRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSS

position 456

SGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTP

(immediately

WGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIAN

subsequent to

NLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS

position 455);

SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLS

modification at

KTINGHDSPHSKAQNHQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNN

positions 454,

SEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVD

455, and 464

ADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDR

underlined;

DVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDK

742 aa

LNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYS

EPRPIGTRYLTRNL

TABLE 5

Exemplary full length capsid nucleic acid sequences

SEQ

Name and
ID

Annotation
NO:
NT Sequence

TTM-001
983
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

9mer peptide

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

underlined

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGACCAATACTTGTAtTActTgagt

AAaACaATTAACGGAAGCGGAAGCCCACACAGCAAAGCACAAAACCAACAGACCtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTaTAA

TTM-002
984
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

7mer peptide

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

underlined

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGACCAATACTTGTAtTActTgagt

AAaACaATTAACGGACACGACAGCCCACACAAAAGCGGACAAAACCAACAGACCtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTaTAA

TTM-023
12
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGGAGCGTGTGTCTGGTTCTCCGCATTCTAAGGCGCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAAcATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-004
13
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGGCGGAGATTGGTCATGATTCTCCGCATAAGTCTGGTCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAAcATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTAATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-005
14
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGATTATTGGTTCTGGTTCTCCGCATTCTAAGGCGCAGAATCGTCATACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-006
15
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGGAGAAGATGTCTGGTTCTCCGCATTCTAAGGCGCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-007
16
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGGAGATTAATGGTCGTGGTTCTCCGCATTCTAAGGCGCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-008
17
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGTTTAATGGTTCTGGTTCTCCGCATTCTAAGGCGCCGAATCTGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-009
18
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGGAGAAGACGTCTGGTTCTCCGCATTCTAAGGCGCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-010
19
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGATGAATGGTCATGATTCTCCGCATTCTAAGGCGCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-011
20
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGATTGATGGTCATGATTCTCCGCATTCTAAGGCGCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-012
21
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGAATAATGGTCATGATTCTCCGCATTCTAAGGCGCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-013
22
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGCAGCGTAAGTCTGGTTCTCCGCATTCTAAGGCGCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-014
23
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGATTAATGGTTCTGGTTCTCCGCATTCTAAGGCGCAGGCGCGTAAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-015
24
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGTATATTGTGGGTTCTGGTTCTCCGCATTCTAAGGCGCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-016
25
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGATTTCTAAGCGTGGTTCTCCGCATTCTAAGGCGCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-017
26
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGGGTCTGGGTGGTTCTGGTTCTCCGCATTCTAAGGCGCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-018
27
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGATTAATGGTCATGATTCTCCGCATTCTAAGGCGCAGAATCTGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-019
28
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGGTGAATGGTCATGATTCTCCGCATTCTAAGGCGCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-020
29
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGATTAATGGTCATGATTCTCCGCATTCTAAGGCGCTGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-021
30
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGATTAATGGTCATGATTCTCCGCATTCTAAGGCGCAGAATCAGCAGTCTtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-022
31
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGATTAATGGTCATGATTCTCCGCATTCTAAGGCGCAGAATCAGCAGATTtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-023
32
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGATTAATGGTTCTGGTTCTCCGCATTTTACGCGTCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-024
33
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGTCTAATGGTCATGATTCTCCGCATTCTAAGGCGCAGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-025
34
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGATTAATGGTTCTGGTTCTCCGCATTCTCTGCCGTGGAATCAGCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

TTM-026
35
ATGGCTGCCGATGGTTATCTTCCAGattggcTCGAGGACAACCTTAGTGAAGGAAT

TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC

ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC

AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA

CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA

ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC

AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT

GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC

CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA

AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC

AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG

GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC

TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG

CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA

ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA

GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT

TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT

AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT

GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA

TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG

TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT

CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC

AAAGCCTGGACCGACTAATGAATCCACTCATCGAtCAgTAtcTGTAtTActTgagt

AAGACGATTAATGGTCATGATTCTCCGCATTCTAAGGCGCAGAATCATCAGACGtT

gAAgTTttcgGTaGCtGGtCCtAGCAACATGGCTGTCCAGGGAAGAAACTACATAC

CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC

AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT

GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC

CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT

GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC

AACGGAGTCCTATGGACAAGtggccacaaaccaccagagtGCCCAAGCACAGGCGC

AGaccggctgggttcaaaaccaAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA

GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT

TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC

TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG

CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG

GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA

ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT

GAACCCCGCCCCATTGGCACgcGgTAttTAACgaGgAActTa

In some embodiments, the polynucleotide encoding an AAV capsid variant, described herein comprises the nucleotide sequence of SEQ ID NO: 983 or 984, or a nucleotide sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.

In some embodiments, the polynucleotide encoding an AAV capsid variant, described herein comprises the nucleotide sequence of any one of SEQ ID NOs: 12-35, or a nucleotide sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.

In some embodiments, the polynucleotide encoding an AAV capsid variant described herein comprises the nucleotide sequence of SEQ ID NO: 983, or a nucleotide sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, the nucleotide sequence encoding an AAV capsid variant described herein, comprises a nucleotide sequence comprising at least one, two or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 983. In some embodiments, the nucleotide sequence encoding an AAV capsid variant described herein, comprises a nucleotide sequence comprising at least one, two or three, but not more than 30, 20 or 10 different nucleotides relative to the amino acid sequence of SEQ ID NO: 983. In some embodiments, the nucleic acid sequence encoding an AAV capsid variant described herein is codon optimized.

In some embodiments, the polynucleotide encoding an AAV capsid variant described herein comprises the nucleotide sequence of SEQ ID NO: 984, or a nucleotide sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, the nucleotide sequence encoding an AAV capsid variant described herein, comprises a nucleotide sequence comprising at least one, two or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 984. In some embodiments, the nucleotide sequence encoding an AAV capsid variant described herein, comprises a nucleotide sequence comprising at least one, two or three, but not more than 30, 20 or 10 different nucleotides, relative to the amino acid sequence of SEQ ID NO: 984. In some embodiments, the nucleic acid sequence encoding an AAV capsid variant described herein is codon optimized.

In some embodiments, an AAV capsid variant described herein comprises the amino acid sequence of any one of SEQ ID NOs: 36-59, 981, or 982, or an amino acid sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of SEQ ID NO: 36-59, 981, or 982. In some embodiments, the AAV capsid variant, comprises an amino acid sequence comprising at least one, two or three, but not more than 30, 20 or 10 different amino acids, relative to the amino acid sequence of SEQ ID NO: 36-59, 981, or 982.

In some embodiments, an AAV capsid variant described herein, comprises the amino acid sequence of SEQ ID NO: 981, or an amino acid sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of SEQ ID NO: 981. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence comprising at least one, two or three, but not more than 30, 20 or 10 different amino acids, relative to the amino acid sequence of SEQ ID NO: 981.

In some embodiments, an AAV capsid variant described herein comprises the amino acid sequence of SEQ ID NO: 982, or an amino acid sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of SEQ ID NO: 982. In some embodiments, the AAV capsid variant, comprises an amino acid sequence comprising at least one, two or three, but not more than 30, 20 or 10 different amino acids, relative to the amino acid sequence of SEQ ID NO: 982.

In some embodiments, an AAV capsid variant described herein comprises the amino acid sequence of SEQ ID NO: 36, or an amino acid sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of SEQ ID NO: 36. In some embodiments, the AAV capsid variant, comprises an amino acid sequence comprising at least one, two or three, but not more than 30, 20 or 10 different amino acids, relative to the amino acid sequence of SEQ ID NO: 36.

In some embodiments, an AAV capsid variant described herein comprises the amino acid sequence of SEQ ID NO: 39, or an amino acid sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of SEQ ID NO: 39. In some embodiments, the AAV capsid variant, comprises an amino acid sequence comprising at least one, two or three, but not more than 30, 20 or 10 different amino acids, relative to the amino acid sequence of SEQ ID NO: 39.

In some embodiments, an AAV capsid variant described herein comprises the amino acid sequence of SEQ ID NO: 51, or an amino acid sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of SEQ ID NO: 51. In some embodiments, the AAV capsid variant, comprises an amino acid sequence comprising at least one, two or three, but not more than 30, 20 or 10 different amino acids, relative to the amino acid sequence of SEQ ID NO: 51.

In some embodiments, an AAV capsid variant described herein comprises the amino acid sequence of SEQ ID NO: 52, or an amino acid sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the amino acid sequence of SEQ ID NO: 52. In some embodiments, the AAV capsid variant, comprises an amino acid sequence comprising at least one, two or three, but not more than 30, 20 or 10 different amino acids, relative to the amino acid sequence of SEQ ID NO: 52.

In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 983 or 984, or a nucleotide sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence encoded by a nucleotide sequence comprising at least one, two or three, but not more than 30, 20 or 10 different nucleotides, relative to the amino acid sequence of SEQ ID NO: 983 or 984. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence encoded by a nucleotide sequence comprising at least one, two or three modifications, e.g., substitutions, insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions, insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 983 or 984.

In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence encoded by the nucleotide sequence of any one of SEQ ID NOs: 12-35, or a nucleotide sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence encoded by a nucleotide sequence comprising at least one, two or three, but not more than 30, 20 or 10 different nucleotides, relative to the amino acid sequence of any one of SEQ ID NOs: 12-35. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence encoded by a nucleotide sequence comprising at least one, two or three modifications, e.g., substitutions, insertions, or deletions, but not more than 30, 20 or 10 modifications, e.g., substitutions, insertions, or deletions, relative to the nucleotide sequence of any one of SEQ ID NOs: 12-35.

In some embodiments, an AAV capsid variant described herein comprises a VP1, VP2, VP3 protein, or a combination thereof. In some embodiments, an AAV capsid variant comprises the amino acid sequence corresponding to positions 138-742, e.g., a VP2, of SEQ ID NO: 981 or 982, or a sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, the AAV capsid protein comprises the amino acid sequence corresponding to positions 203-742, e.g., a VP3, of SEQ ID NO: 981 or 982, or a sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, the AAV capsid variant comprises the amino acid sequence corresponding to positions 1-742, e.g., a VP1, of SEQ ID NO: 981 or 982, or an amino acid sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.

In some embodiments, an AAV capsid variant described herein comprises a VP1, VP2, VP3 protein, or a combination thereof. In some embodiments, an AAV capsid variant comprises the amino acid sequence corresponding to positions 138-742, e.g., a VP2, of any one of SEQ ID NOs: 36-59, or a sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, the AAV capsid protein comprises the amino acid sequence corresponding to positions 203-742, e.g., a VP3, of any one of SEQ ID NOs: 36-59, or a sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto. In some embodiments, the AAV capsid variant comprises the amino acid sequence corresponding to positions 1-742, e.g., a VP1, of any one of SEQ ID NOs: 36-59, or an amino acid sequence with at least 70% (e.g., at least about 80, 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto.

In some embodiments, an AAV capsid variant, described herein has an increased tropism for a CNS cell or tissue, e.g., a brain cell, brain tissue, spinal cord cell, or spinal cord tissue, relative to the tropism of a reference sequence comprising the amino acid sequence of SEQ ID NO: 138.

In some embodiments, an AAV capsid variant described herein transduces a brain region, e.g., a midbrain region (e.g., the hippocampus, or thalamus) or the brain stem. In some embodiments, the level of transduction is at least 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, or 65-fold greater as compared to a reference sequence of SEQ ID NO: 138. In some embodiments, the level of transduction is at least 30, 35, 40, 45, 50, 55, 60, or 65-fold greater as compared to a reference sequence of SEQ ID NO: 138.

In some embodiments, an AAV capsid variant described herein is enriched at least about 3, 4, 5, 6, 7, 8, 9, or 10-fold in the brain compared to a reference sequence of SEQ ID NO: 138. In some embodiments, an AAV capsid variant described herein is enriched at least about 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80 or 85-fold in the brain compared to a reference sequence of SEQ ID NO: 138.

In some embodiments, an AAV capsid variant described herein is enriched in the brain of at least two to three species, e.g., a non-human primate and rodent (e.g., mouse) species, compared to a reference sequence of SEQ ID NO: 138. In some embodiments, an AAV capsid variant described herein is enriched at least about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100-fold in the brain of at least two to three species, e.g., a non-human primate and rodent (e.g., mouse) species, compared to a reference sequence of SEQ ID NO: 138. In some embodiments, the at least two to three species are Macacafascicularis, Chlorocebus sabaeus, Callithrixjacchus, and/or mouse (e.g., BALB/c mice, C57B1/6 mice, and/or CD-1 outbred mice).

In some embodiments, an AAV capsid variant described herein is enriched at least about 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, or 8-fold, in the brain compared to a reference sequence of SEQ ID NO: 981. In some embodiments, an AAV capsid variant described herein is enriched about 2, 2.5, 3, 3.5, 4, 4.5, 5, or 5.5-fold, in the brain compared to a reference sequence of SEQ ID NO: 982.

In some embodiments, an AAV capsid variant described herein delivers an increased level of viral genomes to a brain region. In some embodiments, the level of viral genomes is increased by at least 20, 25, 30, 35, 40, 45, or 50-fold, as compared to a reference sequence of SEQ ID NO: 138. In some embodiments, the brain region comprises a midbrain region (e.g., the hippocampus or thalamus) and/or the brainstem.

In some embodiments, an AAV capsid variant described herein delivers an increased level of a payload to a brain region. In some embodiments, the level of the payload is increased by at least 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, or 70-fold, as compared to a reference sequence of SEQ ID NO: 138. In some embodiments, the brain region comprises a midbrain region (e.g., the hippocampus or thalamus) and/or the brainstem.

In some embodiments, an AAV capsid variant described herein is enriched at least about 5, 10, 15, 20, 25, 30, or 35-fold, in the spinal cord compared to a reference sequence of SEQ ID NO: 138.

In some embodiments, an AAV capsid variant described herein shows preferential transduction in a brain region relative to the transduction in the dorsal root ganglia (DRG). In some embodiments, the AAV capsid variant shows preferential transduction in a brain region relative to the transduction in the liver. In some embodiments, the AAV capsid variant shows preferential transduction in a brain region relative to the transduction in the liver and the DRG. In some embodiments, the AAV capsid variant shows preferential transduction in a brain region relative to the transduction in the heart. In some embodiments, the AAV capsid variant shows preferential transduction in a brain region relative to the transduction in the heart and DRG. In some embodiments, the AAV capsid variant shows preferential transduction in a brain region relative to the transduction in the heart, DRG, and liver. In some embodiments, the AAV capsid variant shows preferential transduction in a brain region and/or a heart region relative to the transduction in the liver and DRG.

In some embodiments, an AAV capsid variant described herein is capable of transducing non-neuronal cells, e.g., glial cells (e.g., oligodendrocytes or astrocytes). In some embodiments, the AAV capsid variant described herein is capable of transducing neuronal cells and non-neuronal cells, e.g., glial cells (e.g., oligodendrocytes or astrocytes). In some embodiments, the non-neuronal cells are glial cells, oligodendrocytes (e.g., Olig2 positive oligodendrocytes), or astrocytes (e.g., Olig2 positive astrocytes). In some embodiments, the AAV capsid variant is capable of transducing Olig2 positive cells, e.g., Olig2 positive astrocytes or Olig2 positive oligodendrocytes.

In some embodiments, an AAV capsid polypeptide, e.g., an AAV capsid variant, described herein has an increased tropism for a muscle cell or tissue, e.g., a heart or quadriceps cell or tissue, relative to the tropism of a reference sequence comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant is enriched at least about 4, 5, 8, 12, 17, 18, 20, 26, 27, 28, 30, or 36-fold, in the muscle compared to a reference sequence of SEQ ID NO: 138. In some embodiments, the muscle region comprises a quadriceps muscle, heart muscle, and/or a diaphragm muscle region. In some embodiments, the muscle region comprises a heart muscle region, e.g., a heart atrium muscle region or a heart ventricle muscle region.

In some embodiments, an AAV capsid polypeptide, e.g., an AAV capsid variant, described herein has an increased tropism for a heart cell or heart tissue. In some embodiments, the AAV capsid variant is enriched at least about 4, 5, 8, 10, 11, 12, 13, 14, 18, 19, 20, 21, 22, 24, 25, 27, 31, 33, or 34-fold, in the heart compared to a reference sequence of SEQ ID NO: 138.

In some embodiments, an AAV capsid variant described herein has increased tropism for a liver cell or tissue, relative to the tropism of a reference sequence comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant is enriched at least about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 150, 160, 170, 180, 190, or 200-fold, in the liver compared to a reference sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant has reduced tropism for a CNS cell or tissue, e.g., a brain cell, brain tissue, spinal cord cell, or spinal cord tissue, relative to the tropism of a reference sequence comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant shows preferential transduction in a liver region relative to the transduction in the brain and/or dorsal root ganglia (DRG). In some embodiments, the AAV capsid variant shows preferential transduction in a liver region relative to the transduction in the heart and/or muscle (e.g., quadriceps).

In some embodiments, an AAV capsid variant of the present disclosure has decreased tropism for the liver. In some embodiments, an AAV capsid variant comprises a modification, e.g., substitution (e.g., conservative substitution), insertion, or deletion, that results in reduced tropism (e.g., de-targeting) and/or activity in the liver. In some embodiments, the reduced tropism in the liver is compared to an otherwise similar capsid that does not comprise the modification, e.g., a wild-type capsid. In some embodiments, an AAV capsid variant described herein comprises a modification, e.g., substitution (e.g., conservative substitution), insertion, or deletion that results in one or more of the following properties: (1) reduced tropism in the liver; (2) reduced, e.g., de-targeted, expression in the liver; (3) reduced activity in the liver; and/or (4) reduced binding to galactose. In some embodiments, the reduction in any one, or all of properties (1)-(3) is compared to an otherwise similar AAV capsid variant that does not comprise the modification. Exemplary modifications are provided in WO 2018/119330; Pulicherla et al. (2011) Mol. Ther. 19(6): 1070-1078; Adachi et al. (2014) Nature Communications 5(3075), DOI: 10.1038/ncomms4075; and Bell et al. (2012) J. Virol. 86(13): 7326-33; the contents of which are hereby incorporated by reference in their entirety. In some embodiments, the AAV capsid variant comprises a modification e.g., substitution (e.g., conservative substitution), insertion, or deletion, at position N470 (e.g., N470A), D271 (e.g., D271A), N272 (e.g., N297A), Y446 (e.g., Y446A), N498 (e.g., N498Y or N4981), W503 (e.g., W530R or W530A), L620 (e.g., L620F), or a combination thereof, relative to a reference sequence numbered according to SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises one, two, three, four, five or all of an amino acid other than N at position 470 (e.g., A), an amino acid other than D at position 271 (e.g., A), an amino acid other than N at position 272 (e.g., A), an amino acid other than Y at position 446 (e.g., A), and amino acid other than N at position 498/(e.g., Y or I), and amino acid other than W at position 503 (e.g., R or A), and amino acid other than L at position 620 (e.g., F), relative to a reference sequence numbered according to SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises a modification e.g., substitution (e.g., conservative substitution), insertion, or deletion, at position N470 (e.g., N470A), D271 (e.g., D271A), N272 (e.g., N297A), Y446 (e.g., Y446A), and W503 (e.g., W530R or W530A), relative to a reference sequence numbered according to SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises a modification e.g., substitution (e.g., conservative substitution), insertion, or deletion, at N498 (e.g., N498Y) and L620 (e.g., L620F).

In some embodiments, an AAV capsid variant comprised herein comprises a modification as described in Adachi et al. (2014) Nature Communications 5(3075), DOI: 10.1038/ncomms4075, the contents of which are hereby incorporated by reference in its entirety. Exemplary modifications that alter or do not alter tissue transduction in at least the brain, liver, heart, lung, and/or kidney can be found in Supplementary Data 2 showing the AAV Barcode-Seq data obtained with AAV9-AA-VBCLib of Adachi et al. (supra), the contents of which are hereby incorporated by reference in its entirety.

In some embodiments, an AAV capsid variant of the present disclosure is isolated, e.g., recombinant. In some embodiments, a polynucleotide encoding an AAV capsid polypeptide, e.g., an AAV capsid variant, of the present disclosure is isolated, e.g., recombinant.

Also provided herein are polynucleotide sequences encoding any of the AAV capsid variants described above and AAV particles, vectors, and cells comprising the same.

AAV Serotypes and Capsids

In some embodiments, an AAV particle of the present disclosure may comprise a capsid protein or variant thereof any natural or recombinant AAV serotype. AAV serotypes may differ in characteristics such as, but not limited to, packaging, tropism, transduction and immunogenic profiles. While not wishing to be bound by theory, it is believed in some embodiments, that the AAV capsid protein, e.g., an AAV capsid variant, can modulate, e.g., direct, AAV particle tropism to a particular tissue.

In some embodiments, an AAV capsid variant described herein allows for blood brain barrier penetration following intravenous administration. In some embodiments, the AAV capsid variant allows for blood brain barrier penetration following intravenous administration, focused ultrasound (FUS), e.g., coupled with the intravenous administration of microbubbles (FUS-MB), or MRI-guided FUS coupled with intravenous administration. In some embodiments the AAV capsid variant allows for increased distribution to a brain region. In some embodiments, the brain region comprises a frontal cortex, sensory cortex, motor cortex, caudate, dentate nucleus, cerebellar cortex, cerebral cortex, brain stem, hippocampus, thalamus, putamen, or a combination thereof. In some embodiments, the AAV capsid variant allows for preferential transduction in a brain region relative to the transduction in the dorsal root ganglia (DRG). In some embodiments, the AAV capsid variant allows for transduction in a non-neuronal cell, e.g., a glial cell (e.g., an astrocyte, an oligodendrocyte, or a combination thereof).

In some embodiments, an AAV capsid variant allows for increased distribution to a spinal cord region. In some embodiments, the spinal region comprises a cervical spinal cord region, thoracic spinal cord region, and/or lumbar spinal cord region.

In some embodiments, the AAV capsid variant, is suitable for intramuscular administration and/or transduction of muscle fibers. In some embodiments the AAV capsid variant, allows for increased distribution to a muscle region. In some embodiments, the muscle region comprises a heart muscle, quadriceps muscle, a diaphragm muscle region, or a combination thereof. In some embodiments, the muscle region comprises a heart muscle region, e.g., a heart atrium muscle region or a heart ventricle muscle region.

In some embodiments, the initiation codon for translation of the AAV VP1 capsid protein, e.g., a capsid variant, described herein may be CTG, TTG, or GTG as described in U.S. Pat. No. 8,163,543, the contents of which are herein incorporated by reference in its entirety. The present disclosure refers to structural capsid proteins (including VP1, VP2 and VP3) which are encoded by capsid (Cap) genes. These capsid proteins form an outer protein structural shell (e.g., capsid) of a viral vector such as AAV. VP capsid proteins synthesized from Cap polynucleotides generally include a methionine as the first amino acid in the peptide sequence (Met1), which is associated with the start codon (AUG or ATG) in the corresponding Cap nucleotide sequence. However, it is common for a first-methionine (Met1) residue or generally any first amino acid (AA1) to be cleaved off after or during polypeptide synthesis by protein processing enzymes such as Met-aminopeptidases. This “Met/AA-clipping” process often correlates with a corresponding acetylation of the second amino acid in the polypeptide sequence (e.g., alanine, valine, serine, threonine, etc.). Met-clipping commonly occurs with VP1 and VP3 capsid proteins but can also occur with VP2 capsid proteins.

Where the Met/AA-clipping is incomplete, a mixture of one or more (one, two or three) VP capsid proteins comprising the viral capsid may be produced, some of which may include a Met1/AA1 amino acid (Met+/AA+) and some of which may lack a Met1/AA1 amino acid as a result of Met/AA-clipping (Met−/AA−). For further discussion regarding Met/AA-clipping in capsid proteins, see Jin, et al. Direct Liquid Chromatography/Mass Spectrometry Analysis for Complete Characterization of Recombinant Adeno-Associated Virus Capsid Proteins. Hum Gene Ther Methods. 2017 Oct. 28(5):255-267; Hwang, et al. N-Terminal Acetylation of Cellular Proteins Creates Specific Degradation Signals. Science. 2010 Feb. 19. 327(5968): 973-977; the contents of which are each incorporated herein by reference in its entirety.

According to the present disclosure, references to capsid proteins, e.g., AAV capsid variants, is not limited to either clipped (Met−/AA−) or unclipped (Met+/AA+) and may, in context, refer to independent capsid proteins, viral capsids comprised of a mixture of capsid proteins, and/or polynucleotide sequences (or fragments thereof) which encode, describe, produce or result in capsid proteins of the present disclosure. A direct reference to a capsid protein or capsid polypeptide (such as VP1, VP2 or VP2) may also comprise VP capsid proteins which include a Met1/AA1 amino acid (Met+/AA+) as well as corresponding VP capsid proteins which lack the Met1/AA1 amino acid as a result of Met/AA-clipping (Met−/AA−).

Further according to the present disclosure, a reference to a specific SEQ ID NO: (whether a protein or nucleic acid) which comprises or encodes, respectively, one or more capsid proteins which include a Met1/AA1 amino acid (Met+/AA+) should be understood to teach the VP capsid proteins which lack the Met1/AA1 amino acid as upon review of the sequence, it is readily apparent any sequence which merely lacks the first listed amino acid (whether or not Met1/AA1).

As a non-limiting example, reference to a VP1 polypeptide sequence which is 736 amino acids in length, and which includes a “Met1” amino acid (Met+) encoded by the AUG/ATG start codon may also be understood to teach a VP1 polypeptide sequence which is 735 amino acids in length, and which does not include the “Met1” amino acid (Met−) of the 736 amino acid Met+ sequence. As a second non-limiting example, reference to a VP1 polypeptide sequence which is 736 amino acids in length, and which includes an “AA1” amino acid (AA1+) encoded by any NNN initiator codon may also be understood to teach a VP1 polypeptide sequence which is 735 amino acids in length, and which does not include the “AA1” amino acid (AA1−) of the 736 amino acid AA1+ sequence.

References to viral capsids formed from VP capsid proteins (such as reference to specific AAV capsid serotypes), can incorporate VP capsid proteins which include a Met1/AA1 amino acid (Met+/AA1+), corresponding VP capsid proteins which lack the Met1/AA1 amino acid as a result of Met/AA1-clipping (Met−/AA1−), and combinations thereof (Met+/AA1+ and Met−/AA1−).

As a non-limiting example, an AAV capsid serotype can include VP1 (Met+/AA1+), VP1 (Met−/AA1−), or a combination of VP1 (Met+/AA1+) and VP1 (Met−/AA1−). An AAV capsid serotype can also include VP3 (Met+/AA1+), VP3 (Met−/AA1−), or a combination of VP3 (Met+/AA1+) and VP3 (Met−/AA1−); and can also include similar optional combinations of VP2 (Met+/AA1) and VP2 (Met−/AA1−).

Additional AAV Sequences

In some embodiments, the AAV capsid variant, comprises immediately subsequent to position 448, 449, 452, 453, 455, numbered relative to SEQ ID NO: 138 or corresponding to equivalent positions in any other AAV serotype (e.g., AAV1, AAV2, AAV3, AAV3b, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAVrh10, AAVrh32.33, AAVrh74, SEQ ID NO: 1, SEQ ID NO: 11, PHP.N, PHP.B, or an AAV serotype as provided in Table 6 of WO 2021/230987 (the contents of which are hereby incorporated by reference in their entirety)), at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 consecutive amino acids of any of amino acid sequence provided in Tables 1, 2A, 2B, 13-19. In some embodiments, the at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 consecutive amino acids of any of amino acid sequence provided in Tables 1, 2A, 2B, 13-19 replaces at least one, two, three, four, five, six, seven, eight, nine, ten, or all of positions K449, T450, I451, N452, G453, 5454, G455, Q456, N457, Q458, and/or Q459, numbered according to SEQ ID NO: 138 or corresponding to equivalent positions in any other AAV serotype (e.g., AAV1, AAV2, AAV3, AAV3b, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAVrh10, AAVrh32.33, AAVrh74, SEQ ID NO: 1, SEQ ID NO: 11, PHP.N, PHP.B, or an AAV serotype as provided in Table 6 of WO 2021/230987 (the contents of which are hereby incorporated by reference in their entirety). In some embodiments, the at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 consecutive amino acids of any of amino acid sequence provided in Tables 1, 2A, 2B, 13-19 replaces positions S454, G455, or both positions S454 and G455, numbered according to SEQ ID NO: 138 or corresponding to equivalent positions in any other AAV serotype (e.g., AAV1, AAV2, AAV3, AAV3b, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAVrh10, AAVrh32.33, AAVrh74, SEQ ID NO: 1, SEQ ID NO: 11, PHP.N, PHP.B, or an AAV serotype as provided in Table 6 of WO 2021/230987 (the contents of which are hereby incorporated by reference in their entirety). In some embodiments, the AAV capsid variant comprises an amino acid other than the wild-type, e.g., native, amino acid, at one, two, three, four, five, six, seven, eight, nine or all of positions T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, and/or Q459, numbered according to SEQ ID NO: 138 or corresponding to equivalent positions in any other AAV serotype (e.g., AAV1, AAV2, AAV3, AAV3b, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAVrh10, AAVrh32.33, AAVrh74, SEQ ID NO: 1, SEQ ID NO: 11, PHP.N, PHP.B, or an AAV serotype as provided in Table 6 of WO 2021/230987 (the contents of which are hereby incorporated by reference in their entirety). In some embodiments, the AAV capsid variant comprises an amino acid other than the wild-type, e.g., native, amino acid, at position S454, G455, or both positions S454 and G455, numbered according to SEQ ID NO: 138 or corresponding to equivalent positions in any other AAV serotype (e.g., AAV1, AAV2, AAV3, AAV3b, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAVrh10, AAVrh32.33, AAVrh74, SEQ ID NO: 1, SEQ ID NO: 11, PHP.N, PHP.B, or an AAV serotype as provided in Table 6 of WO 2021/230987 (the contents of which are hereby incorporated by reference in their entirety)). In some embodiments, the AAV capsid variant comprises a modification, e.g., substitution, at one, two, three, four, five, six, seven, eight, nine, ten or all of positions K449, T450, 1451, N452, G453, S454, G455, Q456, N457, Q458, and/or Q459, numbered according to SEQ ID NO: 138 or corresponding to equivalent positions in any other AAV serotype (e.g., AAV1, AAV2, AAV3, AAV3b, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAVrh10, AAVrh32.33, AAVrh74, SEQ ID NO: 1, SEQ ID NO: 11, PHP.N, PHP.B, or an AAV serotype as provided in Table 6 of WO 2021/230987 (the contents of which are hereby incorporated by reference in their entirety). In some embodiments, the AAV capsid variant comprises a modification, e.g., substitution, at position S454, G455, or both positions S454 and G455, numbered according to SEQ ID NO: 138 or corresponding to equivalent positions in any other AAV serotype (e.g., AAV1, AAV2, AAV3, AAV3b, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAVrh10, AAVrh32.33, AAVrh74, SEQ ID NO: 1, SEQ ID NO: 11, PHP.N, PHP.B, or an AAV serotype as provided in Table 6 of WO 2021/230987 (the contents of which are hereby incorporated by reference in their entirety).

In some embodiments, an AAV capsid polypeptide or AAV capsid variant described herein may comprise a VOY101 capsid polypeptide, an AAVPHP.B (PHP.B) capsid polypeptide, a AAVPHP.N (PHP.N) capsid polypeptide, an AAV1 capsid polypeptide, an AAV2 capsid polypeptide, an AAV5 capsid polypeptide, an AAV9 capsid polypeptide, an AAV9 K449R capsid polypeptide, an AAVrh10 capsid polypeptide, or a functional variant thereof. In some embodiments, the AAV capsid polypeptide, e.g., AAV capsid variant, comprises an amino acid sequence of any of the AAV capsid polypeptides in Table 6, or an amino acid sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto. In some embodiments, the nucleotide sequence encoding the AAV capsid polypeptide comprises any one of the nucleotide sequences in Table 6, or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto.

In some embodiments, an AAV capsid polypeptide or an AAV capsid variant described herein comprises the amino acid sequence of SEQ ID NO: 138 or an amino acid sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto. In some embodiments the AAV capsid polypeptide or the AAV capsid variant, comprises an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than 30, 20, or 10 modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid polypeptide or the AAV capsid variant, comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 137 or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto. In some embodiments, the nucleotide sequence encoding the AAV capsid polypeptide or the AAV capsid variant comprises the nucleotide sequence of SEQ ID NO: 137 or a nucleotide sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto. In some embodiments, the AAV capsid polypeptide or the AAV capsid variant, comprises substitution at position K449, e.g., a K449R substitution, numbered according to SEQ ID NO: 138.

In some embodiments, the AAV capsid polypeptide or the AAV capsid variant, comprises a peptide comprising the amino acid sequence of TLAVPFK (SEQ ID NO: 4680). In some embodiments, the peptide is present immediately subsequent to position 588, relative to a reference sequence numbered according to SEQ ID NO: 138. In some embodiments, the capsid polypeptide comprises the amino acid substitutions of A587D and Q588G, numbered according to SEQ ID NO: 138.

In some embodiments, the AAV capsid polypeptide or the AAV capsid variant comprises the amino acid substitution of K449R, numbered according to SEQ ID NO: 138; and a peptide comprising the amino acid sequence of TLAVPFK (SEQ ID NO: 4680), wherein the peptide is present immediately subsequent to position 588, relative to a reference sequence numbered according to SEQ ID NO: 138.

In some embodiments, the AAV capsid polypeptide or the AAV capsid variant comprises the amino acid substitution of K449R, numbered according to SEQ ID NO: 138; an peptide comprising the amino acid sequence of TLAVPFK (SEQ ID NO: 4680), wherein the insert is present immediately subsequent to position 588, relative to a reference sequence numbered according to SEQ ID NO: 138; and the amino acid substitutions of A587D and Q588G, numbered according to SEQ ID NO: 138.

In some embodiments, the AAV capsid polypeptide or the AAV capsid variant comprises a peptide comprising the amino acid sequence of TLAVPFK (SEQ ID NO: 4680), wherein the insert is present immediately subsequent to position 588, relative to a reference sequence numbered according to SEQ ID NO: 138; and the amino acid substitutions of A587D and Q588G, numbered according to SEQ ID NO: 138.

In some embodiments, the AAV capsid polypeptide or the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 11 or an amino acid sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto. In some embodiments the AAV capsid polypeptide or the AAV capsid variant, comprises an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than 30, 20, or 10 modifications, e.g., substitutions (conservative substitutions), relative to the amino acid sequence of SEQ ID NO: 11, optionally wherein position 449 is not R.

In some embodiments, the AAV capsid polypeptide or AAV capsid variant, comprises the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence substantially identical (e.g., having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, or 99% sequence identity) thereto. In some embodiments the AAV capsid polypeptide or the AAV capsid variant, comprises an amino acid sequence comprising at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than 30, 20, or 10 modifications, e.g., substitutions (e.g., conservative substitutions), relative to the amino acid sequence of SEQ ID NO: 1.

TABLE 6

AAV Sequences

SEQ

ID

Serotype
NO:
Sequence

VQY101
1
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPGNGLD

KGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGGNLGRAVFQ

AKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAKKRLNFGQTGDTE

SVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVI

TTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQR

LINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTSTVQVFTDSDYQLPYVLGSAH

EGCLPPFPADVFMIPQYGYLTLNDGSQAVGRSSFYCLEYFPSQMLRTGNNFQFSYEFENV

PFHSSYAHSQSLDRLMNPLIDQYLYYLSKTINGSGQNQQTLKFSVAGPSNMAVQGRNYIP

GPSYRQQRVSTTVTQNNNSEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGS

LIFGKQGTGRDNVDADKVMITNEEEIKTTNPVATESYGQVATNHQSDGTLAVPFKAQAQT

GWVQNQGILPGMVWQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTP

VPADPPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEF

AVNTEGVYSEPRPIGTRYLTRNL

AAV9/hu.14
11
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPGNGLD

K449R

KGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGGNLGRAVFQ

AKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAKKRLNFGQTGDTE

SVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVI

TTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQR

LINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTSTVQVFTDSDYQLPYVLGSAH

EGCLPPFPADVEMIPQYGYLTLNDGSQAVGRSSFYCLEYFPSQMLRTGNNFQFSYEFENV

PFHSSYAHSQSLDRLMNPLIDQYLYYLSRTINGSGQNQQTLKFSVAGPSNMAVQGRNYIP

GPSYRQQRVSTTVTQNNNSEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGS

LIFGKQGTGRDNVDADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQG

ILPGMVWQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPT

AFNKDKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGV

YSEPRPIGTRYLTRNL

AAV9/hu.14
138
MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHODNARGLVLPGYKYLGPGNGLD

WT

KGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGGNLGRAVFQ

(amino

AKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAKKRLNFGQTGDTE

acid)

SVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVI

TTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQR

LINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTSTVQVFTDSDYQLPYVLGSAH

EGCLPPFPADVFMIPQYGYLTLNDGSQAVGRSSFYCLEYFPSQMLRTGNNFQFSYEFENV

PFHSSYAHSQSLDRLMNPLIDQYLYYLSKTINGSGQNQQTLKFSVAGPSNMAVQGRNYIP

GPSYRQQRVSTTVTQNNNSEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGS

LIFGKQGTGRDNVDADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQG

ILPGMVWQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPT

AFNKDKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGV

YSEPRPIGTRYLTRNL

AAV9/hu.14
137
ATGGCTGCCGATGGTTATCTTCCAGATTGGCTCGAGGACAACCTTAGTGAAGGAATTCGC

WT

GAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAACATCAAGAC

(DNA)

AACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGCAACGGACTCGAC

AAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCACGACAAGGCCTACGAC

CAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACAACCACGCCGACGCCGAGTTC

CAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGCAACCTCGGGCGAGCAGTCTTCCAG

GCCAAAAAGAGGCTTCTTGAACCTCTTGGTCTGGTTGAGGAAGCGGCTAAGACGGCTCCT

GGAAAGAAGAGGCCTGTAGAGCAGTCTCCTCAGGAACCGGACTCCTCCGCGGGTATTGGC

AAATCGGGTGCACAGCCCGCTAAAAAGAGACTCAATTTCGGTCAGACTGGCGACACAGAG

TCAGTCCCAGACCCTCAACCAATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCT

CTTACAATGGCTTCAGGTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGA

GTGGGTAGTTCCTCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATC

ACCACCAGCACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATC

TCCAACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC

TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCAGCGA

CTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCTTCAACATT

CAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAATAACCTTACCAGC

ACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGTGCTCGGGTCGGCTCAC

GAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGATTCCTCAGTACGGGTATCTG

ACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCGTCCTTTTACTGCCTGGAATATTTC

CCGTCGCAAATGCTAAGAACGGGTAACAACTTCCAGTTCAGCTACGAGTTTGAGAACGTA

CCTTTCCATAGCAGCTACGCTCACAGCCAAAGCCTGGACCGACTAATGAATCCACTCATC

GACCAATACTTGTACTATCTCTCAAAGACTATTAACGGTTCTGGACAGAATCAACAAACG

CTAAAATTCAGTGTGGCCGGACCCAGCAACATGGCTGTCCAGGGAAGAAACTACATACCT

GGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAACAGCGAA

TTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTTGATGAATCCT

GGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTCCTTTGTCTGGATCT

TTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGATGCGGACAAAGTCATGATA

ACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGCAACGGAGTCCTATGGACAAGTG

GCCACAAACCACCAGAGTGCCCAAGCACAGGCGCAGACCGGCTGGGTTCAAAACCAAGGA

ATACTTCCGGGTATGGTTTGGCAGGACAGAGATGTGTACCTGCAAGGACCCATTTGGGCC

AAAATTCCTCACACGGACGGCAACTTTCACCCTTCTCCGCTGATGGGAGGGTTTGGAATG

AAGCACCCGCCTCCTCAGATCCTCATCAAAAACACACCTGTACCTGCGGATCCTCCAACG

GCCTTCAACAAGGACAAGCTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGC

GTGGAGATCGAGTGGGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAG

TACACTTCCAACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTA

TATAGTGAACCCCGCCCCATTGGCACCAGATACCTGACTCGTAATCTGTAA

Viral Genome of the AAV Particle

In some embodiments, an AAV particle as described herein comprising an AAV capsid variant described herein, may be used for the delivery of a viral genome to a tissue (e.g., CNS, DRG, and/or muscle). In some embodiments, an AAV particle comprising an AAV capsid variant described herein can be used for delivery of a viral genome to a tissue or cell, e.g., CNS, DRG, or muscle cell or tissue. In some embodiments, an AAV particle of the present disclosure is a recombinant AAV particle. In some embodiments, an AAV particle of the present disclosure is an isolated AAV particle.

The viral genome may encode any payload, such as but not limited to a polypeptide (e.g., a therapeutic polypeptide), an antibody, an enzyme, an RNAi agent and/or components of a gene editing system. In one embodiment, the AAV particles described herein are used to deliver a payload to cells of the CNS, after intravenous delivery. In another embodiment, the AAV particles described herein are used to deliver a payload to cells of the DRG, after intravenous delivery. In some embodiments, the AAV particles described herein are used to deliver a payload to cells of a muscle, e.g., a heart muscle, after intravenous delivery.

In some embodiments, a viral genome of an AAV particle comprising an AAV capsid variant, as described herein, comprises a nucleotide sequence comprising a transgene encoding a payload. In some embodiments, the viral genome comprises an inverted terminal repeat sequence (ITR). In some embodiments, the viral genome comprises two ITR sequences, one at the 5′ end of the viral genome (e.g., 5′ relative to the encoded payload) and one at the 3′ end of the viral genome (e.g., 3′ relative to the encoded payload). In some embodiments, a viral genome of an AAV particle, e.g., an AAV particle comprising an AAV capsid variant described herein, may comprise a regulatory element (e.g., promoter), untranslated regions (UTR), a miR binding site, a polyadenylation sequence (polyA), a filler or stuffer sequence, an intron, and/or a linker sequence, e.g., for enhancing transgene expression.

In some embodiments, the viral genome components are selected and/or engineered for expression of the payload in a target tissue (e.g., CNS, muscle, or DRG).

Viral Genome Component: Inverted Terminal Repeats (ITRs)

In some embodiments, the AAV particle comprising an AAV capsid variant described herein comprises a viral genome comprising an ITR and a transgene encoding a payload. In some embodiments, the viral genome comprises two ITRs. In some embodiments, the two ITRs flank the nucleotide sequence encoding the payload at the 5′ and 3′ ends. In some embodiments, the ITRs function as origins of replication comprising recognition sites for replication. In some embodiments, the ITRs comprise sequence regions which can be complementary and symmetrically arranged. In some embodiments, the ITRs incorporated into viral genomes as described herein may be comprised of naturally occurring polynucleotide sequences or recombinantly derived polynucleotide sequences.

In some embodiments, the ITR may be from the same serotype as the capsid polypeptide, e.g., capsid variant, selected from any of the known serotypes, or a variant thereof. In some embodiments, the ITR may be of a different serotype than the capsid. In some embodiments, the viral genome comprises two ITR sequence regions, wherein the ITRs are of the same serotype as one another. In some embodiments, the viral genome comprises two ITR sequence regions, wherein the ITRs are of different serotypes. Non-limiting examples include zero, one or both of the ITRs having the same serotype as the capsid. In one embodiment both ITRs of the viral genome of the AAV particle are AAV2 ITRs.

Independently, each ITR may be about 100 to about 150 nucleotides in length. An ITR may be about 100-105 nucleotides in length, 106-110 nucleotides in length, 111-115 nucleotides in length, 116-120 nucleotides in length, 121-125 nucleotides in length, 126-130 nucleotides in length, 131-135 nucleotides in length, 136-140 nucleotides in length, 141-145 nucleotides in length or 146-150 nucleotides in length. In one embodiment, the ITRs are 140-142 nucleotides in length. Non-limiting examples of ITR length are 102, 105, 130, 140, 141, 142, 145 nucleotides in length.

Viral Genome Component: Promoters

In some embodiments, viral genome of an AAV particle described herein comprises at least one element to enhance the payload target specificity and expression (See e.g., Powell et al. Viral Expression Cassette Elements to Enhance Transgene Target Specificity and Expression in Gene Therapy, 2015; the contents of which are herein incorporated by reference in their entirety). Non-limiting examples of elements to enhance payload target specificity and expression include promoters, endogenous miRNAs, post-transcriptional regulatory elements (PREs), polyadenylation (Poly A) signal sequences and upstream enhancers (USEs), CMV enhancers and introns.

In some embodiments, an AAV particle comprising an AAV capsid variant described herein comprises a viral genome comprising a nucleic acid comprising a transgene encoding a payload, wherein the transgene is operably linked to a promoter. In some embodiments, the promoter is a species specific promoter, an inducible promoter, a tissue-specific promoter, or a cell cycle-specific promoter (e.g., a promoter as described in Parr et al., Nat. Med. 3:1145-9 (1997); the contents of which are herein incorporated by reference in their entirety).

In some embodiments, the Promoter may be naturally occurring or non-naturally occurring. Non-limiting examples of promoters include those derived from viruses, plants, mammals, or humans. In some embodiments, the promoters may be those derived from human cells or systems. In some embodiments, the promoter may be truncated or mutated, e.g., a promoter variant.

In some embodiments, the promoter is a ubiquitous promoter, e.g., capable of expression in multiple tissues. In some embodiments the promoter is an human elongation factor 1α-subunit (EF1α) promoter, the cytomegalovirus (CMV) immediate-early enhancer and/or promoter, the chicken β-actin (CBA) promoter and its derivative CAG, β glucuronidase (GUSB) promoter, or ubiquitin C (UBC) promoter. In some embodiments, the promoter is a cell or tissue specific promoter, e.g., capable of expression in tissues or cells of the central or peripheral nervous systems, targeted regions within (e.g., frontal cortex), and/or sub-sets of cells therein (e.g., excitatory neurons). In some embodiments, the promoter is a cell-type specific promoters capable of expression of a payload in excitatory neurons (e.g., glutamatergic), inhibitory neurons (e.g., GABA-ergic), neurons of the sympathetic or parasympathetic nervous system, sensory neurons, neurons of the dorsal root ganglia, motor neurons, or supportive cells of the nervous systems such as microglia, glial cells, astrocytes, oligodendrocytes, and/or Schwann cells.

In some embodiments, the promoter is a liver specific promoter (e.g., hAAT, TBG), skeletal muscle specific promoter (e.g., desmin, MCK, C512), B cell promoter, monocyte promoter, leukocyte promoter, macrophage promoter, pancreatic acinar cell promoter, endothelial cell promoter, lung tissue promoter, and/or cardiac or cardiovascular promoter (e.g., aMHC, cTnT, and CMV-MLC2k).

In some embodiments, the promoter is a tissue-specific promoter for payload expression in a tissue or cell of the central nervous system. In some embodiments, the promoter is a synapsin (Syn) promoter, glutamate vesicular transporter (VGLUT) promoter, vesicular GABA transporter (VGAT) promoter, parvalbumin (PV) promoter, sodium channel Na_v1.8 promoter, tyrosine hydroxylase (TH) promoter, choline acetyltransferase (ChaT) promoter, methyl-CpG binding protein 2 (MeCP2) promoter, Ca²⁺/calmodulin-dependent protein kinase II (CaMKII) promoter, metabotropic glutamate receptor 2 (mGluR2) promoter, neurofilament light (NFL) or heavy (NFH) promoter, neuron-specific enolase (NSE) promoter, β-globin minigene nβ2 promoter, preproenkephalin (PPE) promoter, enkephalin (Enk) promoter, and excitatory amino acid transporter 2 (EAAT2) promoter, or a fragment thereof. In some embodiments, the promoter is a cell-type specific promoter capable of expression in an astrocyte, e.g., a glial fibrillary acidic protein (GFAP) promoter and a EAAT2 promoter, or a fragment thereof. In some embodiments, the promoter is a cell-type specific promoter capable of expression in an oligodendrocyte, e.g., a myelin basic protein (MBP) promoter or a fragment thereof.

In some embodiments, the promoter is a GFAP promoter. In some embodiments, the promoter is a synapsin (syn or synl) promoter, or a fragment thereof.

In some embodiments, the promoter comprises an insulin promoter or a fragment thereof.

In some embodiments, the promoter of the viral genome described herein (e.g., comprised within an AAV particle comprising an AAV capsid variant described herein) comprises an EF-1α promoter or variant thereof, e.g., as provided in Table 8. In some embodiments, the EF-1α promoter comprises the nucleotide sequence of any one of SEQ ID NOs: 987, 988, 990, 991, 995, 996, 998-1007 or any one of the sequences provided in Table 8, a nucleotide sequence comprising at least one, two, or three but no more than four modifications, e.g., substitutions, relative to the nucleotide sequence of SEQ ID NOs: 987, 988, 990, 991, 995, 996, 998-1007 or any one of the sequences provided in Table 8, or a nucleotide sequence with at least 70% (e.g., 80, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to any one of SEQ ID NOs: 987, 988, 990, 991, 995, 996, 998-1007 or any one of the sequences provided in Table 8.

TABLE 8

Exemplary Promoter Variants

SEQ ID

Description
Sequences
NO:

EF1a Promoter
CGTGAGGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGA
987

(intron
GAAGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCGGG

underlined)
GTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGG

GAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTT

TGCCGCCAGAACACAGGTAAGTGCCGTGTGTGGTTCCCGCGGGCCTGGCCTCTTT

ACGGGTTATGGCCCTTGCGTGCCTTGAATTACTTCCACCTGGCTGCAGTACGTGA

TTCTTGATCCCGAGCTTCGGGTTGGAAGTGGGTGGGAGAGTTCGAGGCCTTGCGC

TTAAGGAGCCCCTTCGCCTCGTGCTTGAGTTGAGGCCTGGCCTGGGCGCTGGGGC

CGCCGCGTGCGAATCTGGTGGCACCTTCGCGCCTGTCTCGCTGCTTTCGATAAGT

CTCTAGCCATTTAAAATTTTTGATGACCTGCTGCGACGCTTTTTTTCTGGCAAGA

TAGTCTTGTAAATGCGGGCCAAGATCTGCACACTGGTATTTCGGTTTTTGGGGCC

GCGGGCGGCGACGGGGCCCGTGCGTCCCAGCGCACATGTTCGGCGAGGCGGGGCC

TGCGAGCGCGGCCACCGAGAATCGGACGGGGGTAGTCTCAAGCTGGCCGGCCTGC

TCTGGTGCCTGGCCTCGCGCCGCCGTGTATCGCCCCGCCCTGGGCGGCAAGGCTG

GCCCGGTCGGCACCAGTTGCGTGAGCGGAAAGATGGCCGCTTCCCGGCCCTGCTG

CAGGGAGCTCAAAATGGAGGACGCGGCGCTCGGGAGAGCGGGCGGGTGAGTCACC

CACACAAAGGAAAAGGGCCTTTCCGTCCTCAGCCGTCGCTTCATGTGACTCCACG

GAGTACCGGGCGCCGTCCAGGCACCTCGATTAGTTCTCGAGCTTTTGGAGTACGT

CGTCTTTAGGTTGGGGGGAGGGGTTTTATGCGATGGAGTTTCCCCACACTGAGTG

GGTGGAGACTGAAGTTAGGCCAGCTTGGCACTTGATGTAATTCTCCTTGGAATTT

GCCCTTTTTGAGTTTGGATCTTGGTTCATTCTCAAGCCTCAGACAGTGGTTCAAA

GTTTTTTTCTTCCATTTCAGGTGTCGTGA

miniEF1a
GCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAG
988

GGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAG

TGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATA

AGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAACAC

GCGTAAG

Promoter
GCATG

Variant 1

Promoter
GGTGGAGAAGAGCATG
990

Variant 2

Promoter
GTCATCACTGAGGTGGAGAAGAGCATG
991

Variant 3

Promoter
CGTGAG

Variant 4

Promoter
GT

Variant 5

Promoter
GCTCCGGT

Variant 6

Promoter
GCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAG
995

Variant 19
GGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAG

TGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATA

AGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAACAC

AG

Promoter
GCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAG
996

Variant 20
GGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAG

TGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATA

AGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAACAC

GC

Promoter
GTAAG

Variant 7

Promoter
GTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGG
998

Variant 8
AGGGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAA

AGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATA

TAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAAC

ACGCGTAAG

Promoter
GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTT
999

Variant 9
GGGGGGAGGGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCGGGGTAAAC

TGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAAC

CGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGC

CAGAACACGCGTAAG

Promoter
CGTGAGGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGA
1000

Variant 10
GAAGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCGGG

GTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGG

GAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTT

TGCCGCCAGAACACGCGTAAG

Promoter
CGTGAGGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGA
1001

Variant 11
GAAGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCGGG

GTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGG

GAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTT

TGCCGCCAGAACACAG

Promoter
GCATGCGTGAGGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTC
1002

Variant 12
CCCGAGAAGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGC

GCGGGGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGG

TGGGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACGCGTAAG

Promoter
GCATGCGTGAGGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTC
1003

Variant 13
CCCGAGAAGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGC

GCGGGGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGG

TGGGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAG

Promoter
GGTGGAGAAGAGCATGCGTGAGGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACAT
1004

Variant 14
CGCCCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGCCTA

GAGAAGGTGGCGCGGGGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTT

TTTCCCGAGGGTGGGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTC

TTTTTCGCAACGGGTTTGCCGCCAGAACACGCGTAAG

Promoter
GGTGGAGAAGAGCATGCGTGAGGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACAT
1005

Variant 15
CGCCCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGCCTA

GAGAAGGTGGCGCGGGGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTT

TTTCCCGAGGGTGGGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTC

TTTTTCGCAACGGGTTTGCCGCCAGAACACAG

Promoter
GTCATCACTGAGGTGGAGAAGAGCATGCGTGAGGCTCCGGTGCCCGTCAGTGGGC
1006

Variant 16
AGAGCGCACATCGCCCACAGTCCCCGAGAAGITGGGGGGAGGGGTCGGCAATTGA

ACCGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAGTGATGTCGTGTACT

GGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATAAGTGCAGTAGTCGC

CGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAACACGCGTAAG

Promoter
GTCATCACTGAGGTGGAGAAGAGCATGCGTGAGGCTCCGGTGCCCGTCAGTGGGC
1007

Variant 18
AGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCGGCAATTGA

ACCGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAGTGATGTCGTGTACT

GGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATAAGTGCAGTAGTCGC

CGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAACACAG

Viral Genome Component: Untranslated Regions (UTRs)

In some embodiments, wild type untranslated regions (UTRs) of a gene are transcribed but not translated. Generally, the 5′ UTR starts at the transcription start site and ends at the start codon and the 3′ UTR starts immediately following the stop codon and continues until the termination signal for transcription.

Features typically found in abundantly expressed genes of specific target organs (e.g., CNS tissue, muscle, or DRG) may be engineered into UTRs to enhance stability and protein production. As a non-limiting example, a 5′ UTR from mRNA normally expressed in the brain (e.g., huntingtin) may be used in the viral genomes of the AAV particles described herein to enhance expression in neuronal cells or other cells of the central nervous system.

While not wishing to be bound by theory, wild-type 5′ untranslated regions (UTRs) include features which play roles in translation initiation. Kozak sequences, which are commonly known to be involved in the process by which the ribosome initiates translation of many genes, are usually included in 5′ UTRs. Kozak sequences have the consensus CCR(A/G)CCAUGG, where R is a purine (adenine or guanine) three bases upstream of the start codon (ATG), which is followed by another ‘G’.

In one embodiment, the 5′UTR in the viral genome includes a Kozak sequence.

In one embodiment, the 5′UTR in the viral genome does not include a Kozak sequence.

While not wishing to be bound by theory, wild-type 3′ UTRs are known to have stretches of Adenosines and Uridines embedded therein. These AU rich signatures are particularly prevalent in genes with high rates of turnover. Based on their sequence features and functional properties, the AU rich elements (AREs) can be separated into three classes (Chen et al, 1995, the contents of which are herein incorporated by reference in its entirety): Class I AREs, such as, but not limited to, c-Myc and MyoD, contain several dispersed copies of an AUUUA motif within U-rich regions. Class II AREs, such as, but not limited to, GM-CSF and TNF-α, possess two or more overlapping UUAUUUA(U/A)(U/A) nonamers. Class III ARES, such as, but not limited to, c-Jun and Myogenin, are less well defined. These U rich regions do not contain an AUUUA motif Most proteins binding to the AREs are known to destabilize the messenger, whereas members of the ELAV family, most notably HuR, have been documented to increase the stability of mRNA. HuR binds to AREs of all the three classes. Engineering the HuR specific binding sites into the 3′ UTR of nucleic acid molecules will lead to HuR binding and thus, stabilization of the message in vivo.

Introduction, removal or modification of 3′ UTR AU rich elements (AREs) can be used to modulate the stability of a polynucleotide. When engineering specific polynucleotides, e.g., payload regions of viral genomes, one or more copies of an ARE can be introduced to make polynucleotides less stable and thereby curtail translation and decrease production of the resultant protein. Likewise, AREs can be identified and removed or mutated to increase the intracellular stability and thus increase translation and production of the resultant protein.

In one embodiment, the 3′ UTR of the viral genome may include an oligo(dT) sequence for templated addition of a poly-A tail.

In one embodiment, the viral genome may include at least one miRNA seed, binding site or full sequence. microRNAs (or miRNA or miR) are 19-25 nucleotide noncoding RNAs that bind to the sites of nucleic acid targets and down-regulate gene expression either by reducing nucleic acid molecule stability or by inhibiting translation. In some embodiments, a microRNA sequence comprises a seed region, e.g., a sequence in the region of positions 2-8 of the mature microRNA, which has Watson-Crick sequence fully or partially complementarity to the miRNA target sequence of the nucleic acid.

In one embodiment, the viral genome may be engineered to include, alter or remove at least one miRNA binding site, full sequence or seed region.

Any UTR from any gene known in the art may be incorporated into the viral genome of the AAV particle. These UTRs, or portions thereof, may be placed in the same orientation as in the gene from which they were selected or they may be altered in orientation or location. In one embodiment, the UTR used in the viral genome of the AAV particle may be inverted, shortened, lengthened, made with one or more other 5′ UTRs or 3′ UTRs known in the art. As used herein, the term “altered” as it relates to a UTR, means that the UTR has been changed in some way in relation to a reference sequence. For example, a 3′ or 5′ UTR may be altered relative to a wild type or native UTR by the change in orientation or location as taught above or may be altered by the inclusion of additional nucleotides, deletion of nucleotides, swapping or transposition of nucleotides.

In one embodiment, the viral genome of the AAV particle comprises at least one artificial UTR which is not a variant of a wild type UTR.

In one embodiment, the viral genome of the AAV particle comprises UTRs which have been selected from a family of transcripts whose proteins share a common function, structure, feature or property.

Viral Genome Component: Polyadenylation Sequence

The viral genome of the AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant, described herein) may comprise a polyadenylation sequence. In some embodiments, the viral genome of the AAV particle (e.g., an AAV particle comprising an AAV capsid variant, described herein) comprises a polyadenylation sequence between the 3′ end of the nucleotide sequence encoding the payload and the 5′ end of the 3′ITR.

Viral Genome Component: Introns

In some embodiments, the viral genome of the AAV particle as described herein (e.g., an AAV particle comprising an AAV capsid variant), comprises an element to enhance the payload target specificity and expression (See e.g., Powell et al. Viral Expression Cassette Elements to Enhance Transgene Target Specificity and Expression in Gene Therapy, Discov. Med, 2015, 19(102): 49-57; the contents of which are herein incorporated by reference in their entirety), such as an intron. Non-limiting examples of introns include, MVM (67-97 bps), FIX truncated intron 1 (300 bps), β-globin SD/immunoglobulin heavy chain splice acceptor (250 bps), adenovirus splice donor/immunoglobin splice acceptor (500 bps), SV40 late splice donor/splice acceptor (19S/16S) (180 bps) and hybrid adenovirus splice donor/IgG splice acceptor (230 bps).

Viral Genome Component: Stuffer Sequences

In some embodiments, the viral genome of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant), comprises an element to improve packaging efficiency and expression, such as a stuffer or filler sequence. Non-limiting examples of stuffer sequences include albumin and/or alpha-1 antitrypsin. Any known viral, mammalian, or plant sequence may be manipulated for use as a stuffer sequence.

In one embodiment, the stuffer or filler sequence may be from about 100-3500 nucleotides in length. The stuffer sequence may have a length of about 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900 or 3000 nucleotides.

Viral Genome Component: miRNA

In one embodiment, the viral genome comprises a sequence encoding a miRNA to reduce the expression of the payload in a tissue or cell, e.g., the DRG (dorsal root ganglion), or neurons of other ganglia, such as those of the sympathetic or parasympathetic nervous system. In some embodiments, a miRNA, e.g., a miR183, a miR182, and/or miR96, may be encoded in the viral genome to modulate, e.g., reduce the expression, of the viral genome in a DRG neuron. As another non-limiting example, a miR-122 miRNA may be encoded in the viral genome to modulate, e.g., reduce, the expression of the viral genome in the liver. In some embodiments, a miRNA, e.g., a miR-142-3p, may be encoded in the viral genome to modulate, e.g., reduce, the expression, of the viral genome in a cell or tissue of the hematopoietic lineage, including for example immune cells (e.g., antigen presenting cells or APC, including dendritic cells (DCs), macrophages, and B-lymphocytes). In some embodiments, a miRNA, e.g., a miR-1, may be encoded in the viral genome to modulate, e.g., reduce, the expression, of the viral genome in a cell or tissue of the heart.

Viral Genome Component: miR Binding Site

Tissue- or cell-specific expression of the AAV viral particles disclosed herein can be enhanced by introducing tissue- or cell-specific regulatory sequences, e.g., promoters, enhancers, microRNA binding sites, e.g., a detargeting site. Without wishing to be bound by theory, it is believed that an encoded miR binding site can modulate, e.g., prevent, suppress, or otherwise inhibit, the expression of a gene of interest on the viral genome disclosed herein, based on the expression of the corresponding endogenous microRNA (miRNA) or a corresponding controlled exogenous miRNA in a tissue or cell, e.g., a non-targeting cell or tissue. In some embodiments, a miR binding site modulates, e.g., reduces, expression of the payload encoded by a viral genome of an AAV particle described herein in a cell or tissue where the corresponding mRNA is expressed.

In some embodiments, the viral genome of an AAV particle described herein comprises a nucleotide sequence encoding a microRNA binding site, e.g., a detargeting site. In some embodiments, the viral genome of an AAV particle described herein comprises a nucleotide sequence encoding a miR binding site, a microRNA binding site series (miR BSs), or a reverse complement thereof.

In some embodiments, the nucleotide sequence encoding the miR binding site series or the miR binding site is located in the 3′-UTR region of the viral genome (e.g., 3′ relative to the nucleotide sequence encoding a payload), e.g., before the polyA sequence, 5′-UTR region of the viral genome (e.g., 5′ relative to the nucleotide sequence encoding a payload), or both.

In some embodiments, the encoded miR binding site series comprise at least 1-5 copies, e.g., at least 1-3, 2-4, 3-5, 1, 2, 3, 4, 5 or more copies of a miR binding site (miR BS). In some embodiments, all copies are identical, e.g., comprise the same miR binding site. In some embodiments, the miR binding sites within the encoded miR binding site series are continuous and not separated by a spacer. In some embodiments, the miR binding sites within an encoded miR binding site series are separated by a spacer, e.g., a non-coding sequence. In some embodiments, the spacer is about 1 to 6 nucleotides or about 5 to 10 nucleotides, e.g., about 7-8 nucleotides, nucleotides in length. In some embodiments, the spacer coding sequence or reverse complement thereof comprises one or more of (i) GGAT; (ii) CACGTG; (iii) GCATGC, or a repeat of one or more of (i)-(iii). In some embodiments, the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions relative to the nucleotide sequence of GATAGTTA.

In some embodiments, the encoded miR binding site series comprise at least 1-5 copies, e.g., at least 1-3, 2-4, 3-5, 1, 2, 3, 4, 5 or more copies of a miR binding site (miR BS). In some embodiments, at least 1, 2, 3, 4, 5, or all of the copies are different, e.g., comprise a different miR binding site. In some embodiments, the miR binding sites within the encoded miR binding site series are continuous and not separated by a spacer. In some embodiments, the miR binding sites within an encoded miR binding site series are separated by a spacer, e.g., a non-coding sequence. In some embodiments, the spacer is about 1 to 6 nucleotides or about 5 to 10 nucleotides, e.g., about 7-8 nucleotides, in length. In some embodiments, the spacer comprises one or more of (i) GGAT; (ii) CACGTG; (iii) GCATGC, or a repeat of one or more of (i)-(iii). In some embodiments, the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions relative to the nucleotide sequence of GATAGTTA.

In some embodiments, the encoded miR binding site is substantially identical (e.g., at least 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100% identical), to the miR in the host cell. In some embodiments, the encoded miR binding site comprises at least 1, 2, 3, 4, or 5 mismatches or no more than 6, 7, 8, 9, or 10 mismatches to a miR in the host cell. In some embodiments, the mismatched nucleotides are contiguous. In some embodiments, the mismatched nucleotides are non-contiguous. In some embodiments, the mismatched nucleotides occur outside the seed region-binding sequence of the miR binding site, such as at one or both ends of the miR binding site. In some embodiments, the miR binding site is 100% identical to the miR in the host cell.

In some embodiments, the nucleotide sequence encoding the miR binding site is substantially complementary (e.g., at least 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100% complementary), to the miR in the host cell. In some embodiments, to complementary sequence of the nucleotide sequence encoding the miR binding site comprises at least 1, 2, 3, 4, or 5 mismatches or no more than 6, 7, 8, 9, or 10 mismatches to a miR in the host cell. In some embodiments, the mismatched nucleotides are contiguous. In some embodiments, the mismatched nucleotides are non-contiguous. In some embodiments, the mismatched nucleotides occur outside the seed region-binding sequence of the miR binding site, such as at one or both ends of the miR binding site. In some embodiments, the encoded miR binding site is 100% complementary to the miR in the host cell.

In some embodiments, an encoded miR binding site or sequence region is at least about 10 to about 125 nucleotides in length, e.g., at least about 10 to 50 nucleotides, 10 to 100 nucleotides, 50 to 100 nucleotides, 50 to 125 nucleotides, or 100 to 125 nucleotides in length. In some embodiments, an encoded miR binding site or sequence region is at least about 7 to about 28 nucleotides in length, e.g., at least about 8-28 nucleotides, 7-28 nucleotides, 8-18 nucleotides, 12-28 nucleotides, 20-26 nucleotides, 22 nucleotides, 24 nucleotides, or 26 nucleotides in length, and optionally comprises at least one consecutive region (e.g., 7 or 8 nucleotides) complementary (e.g., fully or partially complementary) to the seed sequence of a miRNA (e.g., a miR122, a miR142, a miR183, or a miR1).

In some embodiments, the encoded miR binding site is complementary (e.g., fully or partially complementary) to a miR expressed in liver or hepatocytes, such as miR122. In some embodiments, the encoded miR binding site or encoded miR binding site series comprises a miR122 binding site sequence. In some embodiments, the encoded miR122 binding site comprises the nucleotide sequence of ACAAACACCATTGTCACACTCCA (SEQ ID NO: 4673), or a nucleotide sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, at least 95%, at least 99%, or 100% sequence identity, or having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., insertions, deletions, or substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of SEQ ID NO: 4673, e.g., wherein the modification can result in a mismatch between the encoded miR binding site and the corresponding miRNA. In some embodiments, the viral genome comprises at least 2, 3, 4, or 5 copies of the encoded miR122 binding site, e.g., an encoded miR122 binding site series, optionally wherein the encoded miR122 binding site series comprises the nucleotide sequence of: ACAAACACCATTGTCACACTCCACACAAACACCATTGTCACACTCCACACAAACACCATTGTCACACT CCA (SEQ ID NO: 4674), or a nucleotide sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, at least 95%, at least 99%, or 100% sequence identity, or having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 4674, e.g., wherein the modification can result in a mismatch between the encoded miR binding site and the corresponding miRNA. In some embodiments, at least two of the encoded miR122 binding sites are connected directly, e.g., without a spacer. In other embodiments, at least two of the encoded miR122 binding sites are separated by a spacer, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides in length, which is located between two or more consecutive encoded miR122 binding site sequences. In embodiments, the spacer is about 1 to 6 nucleotides or about 5 to 10 nucleotides, e.g., about 7-8, in length. In some embodiments, the spacer coding sequence or reverse complement thereof comprises one or more of (i) GGAT; (ii) CACGTG; (iii) GCATGC, or a repeat of one or more of (i)-(iii). In some embodiments, an encoded miR binding site series comprises at least 3-5 copies (e.g., 4 copies) of a miR122 binding site, with or without a spacer, wherein the spacer is about 1 to 6 nucleotides or about 5 to 10 nucleotides, e.g., about 7-8 nucleotides or about 8 nucleotides, in length. In some embodiments, the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions relative to the nucleotide sequence of GATAGTTA.

In some embodiments, the encoded miR binding site is complementary (e.g., fully or partially complementary) to a miR expressed in the heart. In embodiments, the encoded miR binding site or encoded miR binding site series comprises a miR-1 binding site. In some embodiments, the encoded miR-1 binding site comprises the nucleotide sequence of ATACATACTTCTTTACATTCCA (SEQ ID NO: 4679), a nucleotide sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, at least 95%, at least 99%, or 100% sequence identity, or having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 4679, e.g., wherein the modification can result in a mismatch between the encoded miR binding site and the corresponding miRNA. In some embodiments, the viral genome comprises at least 2, 3, 4, or 5 copies of the encoded miR-1 binding site, e.g., an encoded miR-1 binding site series. In some embodiments, the at least 2, 3, 4, or 5 copies (e.g., 2 or 3 copies) of the encoded miR-1 binding site are continuous (e.g., not separated by a spacer) or separated by a spacer. In some embodiments, the spacer is about 1 to 6 nucleotides or about 5 to 10 nucleotides, e.g., about 7-8 nucleotides or about 8 nucleotides, in length. In some embodiments, the spacer sequence comprises one or more of (i) GGAT; (ii) CACGTG; (iii) GCATGC, or a repeat of one or more of (i)-(iii). In some embodiments, the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of GATAGTTA.

In some embodiments, the encoded miR binding site is complementary (e.g., fully or partially complementary) to a miR expressed in hematopoietic lineage, including immune cells (e.g., antigen presenting cells or APC, including dendritic cells (DCs), macrophages, and B-lymphocytes). In some embodiments, the encoded miR binding site complementary to a miR expressed in hematopoietic lineage comprises a nucleotide sequence disclosed, e.g., in US 2018/0066279, the contents of which are incorporated by reference herein in its entirety.

In embodiments, the encoded miR binding site or encoded miR binding site series comprises a miR-142-3p binding site sequence. In some embodiments, the encoded miR-142-3p binding site comprises the nucleotide sequence of TCCATAAAGTAGGAAACACTACA (SEQ ID NO: 4675), a nucleotide sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, at least 95%, at least 99%, or 100% sequence identity, or having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 4675, e.g., wherein the modification can result in a mismatch between the encoded miR binding site and the corresponding miRNA. In some embodiments, the viral genome comprises at least 2, 3, 4, or 5 copies of the encoded miR-142-3p binding site, e.g., an encoded miR-142-3p binding site series. In some embodiments, the at least 2, 3, 4, or 5 copies (e.g., 2 or 3 copies) of the encoded miR-142-3p binding site are continuous (e.g., not separated by a spacer) or separated by a spacer. In some embodiments, the spacer is about 1 to 6 nucleotides or about 5 to 10 nucleotides, e.g., about 7-8 nucleotides or about 8 nucleotides, in length. In some embodiments, the spacer sequence comprises one or more of (i) GGAT; (ii) CACGTG; (iii) GCATGC, or a repeat of one or more of (i)-(iii). In some embodiments, the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of GATAGTTA.

In some embodiments, the encoded miR binding site is complementary (e.g., fully complementary or partially complementary) to a miR expressed in a DRG (dorsal root ganglion) neuron, e.g., a miR183, a miR182, and/or miR96 binding site. In some embodiments, the encoded miR binding site is complementary to a miR expressed in expressed in a DRG neuron comprises a nucleotide sequence disclosed, e.g., in WO2020/132455, the contents of which are incorporated by reference herein in its entirety.

In some embodiments, the encoded miR binding site or encoded miR binding site series comprises a miR183 binding site sequence. In some embodiments, the encoded miR183 binding site comprises the nucleotide sequence of AGTGAATTCTACCAGTGCCATA (SEQ ID NO: 4676), or a nucleotide sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, at least 95%, at least 99%, or 100% sequence identity, or having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 4676, e.g., wherein the modification can result in a mismatch between the encoded miR binding site and the corresponding miRNA. In some embodiments, the sequence complementary to the seed sequence corresponds to the double underlined of the encoded miR-183 binding site sequence. In some embodiments, the viral genome comprises at least comprises at least 2, 3, 4, or 5 copies (e.g., at least 2 or 3 copies) of the encoded miR183 binding site, e.g., an encoded miR183 binding site. In some embodiments, the at least 2, 3, 4, or 5 copies (e.g., 2 or 3 copies) of the encoded miR183 binding site are continuous (e.g., not separated by a spacer) or separated by a spacer. In some embodiments, the spacer is about 1 to 6 nucleotides or about 5 to 10 nucleotides, e.g., about 7-8 nucleotides or about 8 nucleotides, in length. In some embodiments, the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of GATAGTTA. In some embodiments, the spacer sequence comprises one or more of (i) GGAT; (ii) CACGTG; (iii) GCATGC, or a repeat of one or more of (i)-(iii).

In some embodiments, the encoded miR binding site or the encoded miR binding site series comprises a miR182 binding site sequence. In some embodiments, the encoded miR182 binding site comprises, the nucleotide sequence of AGTGTGAGTTCTACCATTGCCAAA (SEQ ID NO: 4677), a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, at least 95%, at least 99%, or 100% sequence identity, or having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 4677, e.g., wherein the modification can result in a mismatch between the encoded miR binding site and the corresponding miRNA. In some embodiments, the viral genome comprises at least 2, 3, 4, or 5 copies of the encoded miR182 binding site, e.g., an encoded miR182 binding site series. In some embodiments, the at least 2, 3, 4, or 5 copies (e.g., 2 or 3 copies) of the encoded miR182 binding site are continuous (e.g., not separated by a spacer) or separated by a spacer. In some embodiments, the spacer is about 1 to 6 nucleotides or about 5 to 10 nucleotides, e.g., about 7-8 nucleotides or about 8 nucleotides, in length. In some embodiments, the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of GATAGTTA. In some embodiments, the spacer sequence comprises one or more of (i) GGAT; (ii) CACGTG; (iii) GCATGC, or a repeat of one or more of (i)-(iii).

In certain embodiments, the encoded miR binding site or the encoded miR binding site series comprises a miR96 binding site sequence. In some embodiments, the encoded miR96 binding site comprises the nucleotide sequence of AGCAAAAATGTGCTAGTGCCAAA (SEQ ID NO: 4678), a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, at least 95%, at least 99%, or 100% sequence identity, or having at least one, two, three, four, five, six, or seven modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than ten modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of SEQ ID NO: 4678, e.g., wherein the modification can result in a mismatch between the encoded miR binding site and the corresponding miRNA. In some embodiments, the viral genome comprises at least 2, 3, 4, or 5 copies of the encoded miR96 binding site, e.g., an encoded miR96 binding site series. In some embodiments, the at least 2, 3, 4, or 5 copies (e.g., 2 or 3 copies) of the encoded miR96 binding site are continuous (e.g., not separated by a spacer) or separated by a spacer. In some embodiments, the spacer is about 1 to 6 nucleotides or about 5 to 10 nucleotides, e.g., about 7-8 nucleotides or about 8 nucleotides, in length. In some embodiments, the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of GATAGTTA. In some embodiments, the spacer sequence comprises one or more of (i) GGAT; (ii) CACGTG; (iii) GCATGC, or a repeat of one or more of (i)-(iii).

In some embodiments, the encoded miR binding site series comprises a miR122 binding site, a miR-1, a miR142 binding site, a miR183 binding site, a miR182 binding site, a miR 96 binding site, or a combination thereof. In some embodiments, the encoded miR binding site series comprises at least 2, 3, 4, or 5 copies of a miR122 binding site, a miR142 binding site, a miR183 binding site, a miR182 binding site, a miR 96 binding site, or a combination thereof. In some embodiments, at least two of the encoded miR binding sites are connected directly, e.g., without a spacer. In other embodiments, at least two of the encoded miR binding sites are separated by a spacer, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides in length, which is located between two or more consecutive encoded miR binding site sequences. In embodiments, the spacer is at least about 5 to 10 nucleotides, e.g., about 7-8 nucleotides or about 8 nucleotides, in length. In some embodiments, the spacer coding sequence or reverse complement thereof comprises one or more of (i) GGAT; (ii) CACGTG; (iii) GCATGC, or a repeat of one or more of (i)-(iii). In some embodiments, the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of GATAGTTA.

In some embodiments, an encoded miR binding site series comprises at least 2-5 copies (e.g., 2 or 3 copies) of a combination of at least two, three, four, five, or all of a miR-1, miR122 binding site, a miR142 binding site, a miR183 binding site, a miR182 binding site, a miR96 binding site, wherein each of the miR binding sites within the series are continuous (e.g., not separated by a spacer) or are separated by a spacer. In some embodiments, the spacer is about 1 to 6 nucleotides or about 5 to 10 nucleotides, e.g., about 7-8 nucleotides or about 8 nucleotides, in length. In some embodiments, the spacer sequence comprises one or more of (i) GGAT; (ii) CACGTG; (iii) GCATGC, or a repeat of one or more of (i)-(iii). In some embodiments, the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of GATAGTTA.

In some embodiments, an encoded miR binding site series comprises at least 2-5 copies (e.g., 2 or 3 copies) of a combination of a miR-122 binding site and a miR-1 binding site, wherein each of the miR binding sites within the series are continuous (e.g., not separated by a spacer) or are separated by a spacer. In some embodiments, the spacer is about 1 to 6 nucleotides or about 5 to 10 nucleotides, e.g., about 7-8 nucleotides or about 8 nucleotides, in length. In some embodiments, the spacer sequence comprises one or more of (i) GGAT; (ii) CACGTG; (iii) GCATGC, or a repeat of one or more of (i)-(iii). In some embodiments, the spacer comprises the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, or three modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, but no more than four modifications, e.g., substitutions (e.g., conservative substitutions), insertions, or deletions, relative to the nucleotide sequence of GATAGTTA.

Genome Size

In one embodiment, the AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant), may comprise a single-stranded or double-stranded viral genome. The size of the viral genome may be small, medium, large or the maximum size. As described above, the viral genome may comprise a promoter and a polyA tail.

In one embodiment, the viral genome may be a small single stranded viral genome. A small single stranded viral genome may be 2.1 to 3.5 kb in size such as, but not limited to, about 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, and 3.5 kb in size.

In one embodiment, the viral genome may be a small double stranded viral genome. A small double stranded viral genome may be 1.3 to 1.7 kb in size such as, but not limited to, about 1.3, 1.4, 1.5, 1.6, and 1.7 kb in size.

In one embodiment, the viral genome may be a medium single stranded viral genome. A medium single stranded viral genome may be 3.6 to 4.3 kb in size such as, but not limited to, about 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2 and 4.3 kb in size.

In one embodiment, the viral genome may be a medium double stranded viral genome. A medium double stranded viral genome may be 1.8 to 2.1 kb in size such as, but not limited to, about 1.8, 1.9, 2.0, and 2.1 kb in size.

In one embodiment, the viral genome may be a large single stranded viral genome. A large single stranded viral genome may be 4.4 to 6.0 kb in size such as, but not limited to, about 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9 and 6.0 kb in size.

In one embodiment, the viral genome may be a large double stranded viral genome. A large double stranded viral genome may be 2.2 to 3.0 kb in size such as, but not limited to, about 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9 and 3.0 kb in size.

Payloads

In some embodiments, an AAV particle of the present disclosure (e.g. an AAV particle comprising an AAV capsid variant described herein) comprises a viral genome comprising a nucleic acid encoding a payload. In some embodiments, the encoded payload is an RNAi agent or a polypeptide. A payload of the present disclosure may be, but is not limited to, a peptide, a polypeptide, a protein, an antibody, an RNAi agent, etc.

In some embodiments, the nucleotide sequence encoding a payload may comprise a combination of coding and non-coding nucleic acid sequences. In some embodiments, the nucleotide sequence encoding the payload may encode a coding or non-coding RNA.

In some embodiments, the AAV particles described herein, e.g., an AAV particle comprising an AAV capsid variant, comprises a nucleic acid encoding a payload. In some embodiments, the encoded payload comprises a therapeutic protein, an antibody, an enzyme, one or more components of a genome editing system, and/or an RNAi agent (e.g., a dsRNA, siRNA, shRNA, pre-miRNA, pri-miRNA, miRNA, stRNA, lncRNA, piRNA, or snoRNA). In some embodiments, the encoded payload modulates, e.g., increases or decreases, the presence, level, and/or activity of a gene, mRNA, protein, or a combination thereof, e.g., in a cell or a tissue.

Polypeptides

In some embodiments, the encoded payload of AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant, described herein comprises a polypeptide, protein, or peptide, e.g., a polypeptide, protein, or peptide described herein. The nucleic acid encoding the payload, may encode a product of any known gene and/or a recombinant version thereof. In some embodiments, the nucleic acid encoding the payload may encode at least one allele of apolipoprotein E (APOE) such as, but not limited to ApoE2, ApoE3 and/or ApoE4. In one embodiment, the nucleic acid encoding the payload encodes ApoE2 (cys112, cys158) protein or a fragment or variant thereof. In one embodiment, the nucleic acid encoding the payload encodes an ApoE3 (cys112, arg158) protein or fragment or variant thereof. In one embodiment, the nucleic acid encoding the payload encodes ApoE4 (arg112, arg158). As another non-limiting example, the encoded payload comprises an aromatic L-amin acid decarboxylase (AADC) protein. As another non-limiting example, the encoded payload comprises an antibody, or a fragment thereof. As another non-limiting example, the encoded payload comprises a human survival of motor neuron (SMN) 1 or SMN2 protein, or fragments or variants thereof. As another non-limiting example, the encoded payload region comprises a glucocerebrosidase (GBA1) protein, or a fragment or variant thereof. As another non-limiting example, the encoded payload comprises a granulin precursor or progranulin (GRN) protein, or a fragment or variant thereof. As another non-limiting example, the encoded payload comprises an aspartoacylase (ASPA) protein, or a fragment or variant thereof. As another non-limiting example, the encoded payload comprises a tripeptidyl peptidase I (CLN2) protein, or a fragment or variant thereof. As another non-limiting example, the encoded payload comprises a beta-galactosidase (GLB1) protein, or a fragment or variant thereof. As another non-limiting example, the encoded payload comprises a N-sulphoglucosamine sulphohydrolase (SGSH) protein, or a fragment or variant thereof. As another non-limiting example, the encoded payload comprises an N-acetyl-alpha-glucosaminidase (NAGLU) protein, or a fragment or variant thereof. As another non-limiting example, the encoded payload comprises a iduronate 2-sulfatase (IDS) protein, or a fragment or variant thereof. As another non-limiting example, the encoded payload comprises an intracellular cholesterol transporter (NPC1) protein, or a fragment or variant thereof. As another non-limiting example, the encoded payload comprises a gigaxonin (GAN) protein, or a fragment or variant thereof. The AAV viral genomes encoding polypeptides described herein may be useful in the fields of human disease, viruses, infections veterinary applications and a variety of in vivo and in vitro settings.

Amino acid sequences of a payload polypeptide encoded by a viral genome described herein, may be translated as a whole polypeptide, a plurality of polypeptides or fragments of polypeptides, which independently may be encoded by one or more nucleic acids, fragments of nucleic acids or variants of any of the aforementioned.

Antibodies and Antibody Binding Fragments

In some embodiments, the encoded payload of AAV particle comprising an AAV capsid variant described herein comprises an antibody or antibody binding fragment. In some embodiments, the antibody may be a full antibody, a fragment, or any functional variant thereof. As non-limiting examples, an antibody may be a native antibody (e.g., with two heavy and two light chains), a heavy chain variable region, a light chain variable region, a heavy chain constant region, a light chain constant region, Fab, Fab′, F(ab′)₂, Fv, or scFv fragments, a diabody, a linear antibody, a single-chain antibody, a multi-specific antibody, an intrabody, one or more heavy chain complementarity determining regions (CDR), one or more light chain CDRs, a bi-specific antibody, a monoclonal antibody, a polyclonal antibody, a humanized antibody, an antibody mimetic, an antibody variant, a miniaturized antibody, a unibody, a maxibody, and/or a chimeric antigen receptor. The encoded antibody or antibody binding fragment may be useful in the treatment of a neurological disease, a neurodegenerative disorder, a muscular disease, a neuromuscular disorder, a neuro-oncological disorder, or any disorder associated with the central and/or peripheral nervous systems.

In some embodiments, the viral genome of the AAV particle (e.g., an AAV particle comprising an AAV capsid variant described herein) may comprise a nucleic acid which has been engineered to enable or enhance the expression of an antibody, or antibody binding fragment thereof.

In some embodiments, the encoded antibody of the payload of an AAV particle comprising an AAV capsid variant, described herein comprises at least one immunoglobulin variable domain sequence. An antibody may include, for example, full-length, mature antibodies and antigen-binding fragments of an antibody. For example, an antibody can include a heavy (H) chain variable domain sequence (VH), and a light (L) chain variable domain sequence (VL). In another example, an antibody includes two heavy (H) chain variable domain sequences and two light (L) chain variable domain sequence, thereby forming two antigen binding sites, such as Fab, Fab′, F(ab′)2, Fc, Fd, Fd′, Fv, single chain antibodies (scFv for example), single variable domain antibodies, diabodies (Dab) (bivalent and bispecific), and chimeric (e.g., humanized) antibodies, which may be produced by the modification of whole antibodies or those synthesized de novo using recombinant DNA technologies. These functional antibody fragments, e.g., an antibody binding fragments, retain the ability to selectively bind with their respective antigen or receptor.

In some embodiments, the antibody binding fragment comprises at least one portion of an intact antibody, or recombinant variants thereof, and refers to the antigen binding domain, for example, an antigenic determining variable region of an intact antibody, that is sufficient to confer recognition and specific binding of the antibody fragment to a target, such as an antigen. Examples of antigen binding fragments include: (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab′)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a diabody (dAb) fragment, which consists of a VH domain; (vi) a camelid or camelized variable domain; (vii) a single chain Fv (scFv), see e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883); and (viii) a single domain antibody. These antibody fragments are obtained using conventional techniques known to those with skill in the art, and the fragments are screened for utility in the same manner as are intact antibodies. An antibody fragment can also be incorporated into single domain antibodies, maxibodies, minibodies, nanobodies, intrabodies, diabodies, triabodies, tetrabodies, v-NAR and bis-scFv (see, for example, Hollinger and Hudson, Nature Biotechnology 23:1126-1136, 2005).

In some embodiments, the encoded antibody of the payload of an AAV particle described herein comprises a multispecific antibody, e.g., it comprises a plurality of immunoglobulin variable domains sequences, wherein a first immunoglobulin variable domain sequence of the plurality has binding specificity for a first epitope and a second immunoglobulin variable domain sequence of the plurality has binding specificity for a second epitope. In some embodiments, the first and second epitopes are on the same antigen, e.g., the same protein (or subunit of a multimeric protein). In some embodiments, the first and second epitopes overlap. In some embodiments, the first and second epitopes do not overlap. In some embodiments, the first and second epitopes are on different antigens, e.g., the different proteins (or different subunits of a multimeric protein). In some embodiments, a multispecific antibody comprises a third, fourth or fifth immunoglobulin variable domain. In some embodiments, a multispecific antibody is a bispecific antibody, a trispecific antibody, or tetraspecific antibody.

In some embodiments, an encoded multispecific antibody of the payload of an AAV particle described herein is an encoded bispecific antibody. A bispecific antibody has specificity for no more than two antigens. A bispecific antibody is characterized by a first immunoglobulin variable domain sequence which has binding specificity for a first epitope and a second immunoglobulin variable domain sequence that has binding specificity for a second epitope. In some embodiments, the first and second epitopes are on the same antigen, e.g., the same protein (or subunit of a multimeric protein). In some embodiments, the first and second epitopes overlap. In some embodiments, the first and second epitopes do not overlap. In some embodiments, the first and second epitopes are on different antigens, e.g., the different proteins (or different subunits of a multimeric protein).

An antibody or an antibody binding fragment encoded by a viral genome of an AAV particle described herein, may be, but is not limited to, an antibody or antibody fragment that binds to β3-amyloid, APOE, tau, SOD1, TDP-43, huntingtin, and/or synuclein. In some embodiments, the encoded payload comprises an antibody or antibody fragment that binds to a neuro-oncology related target, e.g., HER2, EGFR (e.g., EGFRvIII). In some embodiments, the encoded payload comprises an antibody that binds to HER2/neu. In some embodiments, the encoded payload comprises an antibody that binds to β3-amyloid. In some embodiments, the encoded payload comprises an antibody that binds to tau.

Gene Editing System

In some embodiments, the encoded payload of AAV particle comprising an AAV capsid variant described herein comprises a gene editing system or one or more components thereof. In some embodiments, the gene editing system comprises nucleic acid sequences that encode proteins having enzymatic activity to (i) selectively induce double or single stranded breaks in a DNA or RNA sequence, or (ii) substitute, insert or delete a particular base or set of bases of a DNA or RNA sequence in the absence of a double or single stranded break in the DNA or RNA. In some embodiments, the gene editing system includes, but is not limited to a CRISPR-Cas system (including different Cas or Cas-related nucleases), a Zinc finger nuclease, a meganuclease, a TALEN or a base editors. In some embodiments, the gene editing system comprises a chromosomal integration of a transgene, e.g., introduced by a parvovirus vector in the absence of an exogenous nuclease or an enzymatic entity.

RNAi Agents

In some embodiments, the encoded payload of AAV particle comprising an AAV capsid variant described herein comprises an RNAi agent, e.g., an RNAi agent described herein. In some embodiments, the encoded payload of a viral genome of an AAV particle comprising an AAV capsid variant described herein comprises a dsRNA, a siRNA, a shRNA, a pre-miRNA, a pri-miRNA, a miRNA, a stRNA, a lncRNA, a piRNA, or a snoRNA. In some embodiments, the encoded payload comprises an RNAi agent for inhibiting expression of a SOD1, MAPT, APOE, HTT, C90RF72, TDP-43, APP, BACE, SNCA, ATXN1, ATXN3, ATXN7, SCN1A-SCN5A, or SCN8A-SCN11A gene, protein, and/or mRNA. In some embodiments, the RNAi agent encoded by a viral genome described herein inhibits SOD1, MAPT, APOE, HTT, C90RF72, TDP-43, APP, BACE, SNCA, ATXN1, ATXN3, ATXN7, SCN1A-SCN5A, or SCN8A-SCN11A.

An AAV particle comprising an AAV capsid variant described herein may comprise a viral genome encoding an RNAi agent, which targets the mRNA of a gene to modulate, e.g., interfere with gene expression and/or protein production.

In some embodiments, the RNAi agent may target a gene at the location of a single-nucleotide polymorphism (SNP) or variant within the nucleotide sequence of the gene.

The RNAi agent may be an siRNA duplex, wherein the siRNA duplex contains an antisense strand (guide strand) and a sense strand (passenger strand) hybridized together forming a duplex structure, wherein the antisense strand is complementary to the nucleic acid sequence of the targeted gene, and wherein the sense strand is homologous to the nucleic acid sequence of the targeted gene. In some aspects, the 5′ end of the antisense strand has a 5′ phosphate group and the 3′ end of the sense strand contains a 3′ hydroxyl group. In other aspects, there are none, one or 2 nucleotide overhangs at the 3′ end of each strand.

Each strand of an siRNA duplex targeting a gene of interest may be about 19 to 25, 19 to 24 or 19 to 21 nucleotides in length, preferably about 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, or 25 nucleotides in length.

In one embodiment, an siRNA or dsRNA includes at least two sequences that are complementary to each other. The dsRNA includes a sense strand having a first sequence and an antisense strand having a second sequence. The antisense strand includes a nucleotide sequence that is substantially complementary to at least part of an mRNA encoding the target gene, and the region of complementarity is 30 nucleotides or less, and at least 15 nucleotides in length. Generally, the dsRNA is 19 to 25, 19 to 24 or 19 to 21 nucleotides in length. In some embodiments, the dsRNA is from about 15 to about 25 nucleotides in length, and in other embodiments the dsRNA is from about 25 to about 30 nucleotides in length. In some embodiments, the dsRNA is about 15 nucleotides in length, 16 nucleotides in length, 17 nucleotides in length, 18 nucleotides in length, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides in length, 26 nucleotides in length, 27 nucleotides in length, 28 nucleotides in length, 29 nucleotides in length, or 30 nucleotides in length.

In some embodiments, the encoded RNAi agent is an siRNA.

In some embodiments, the RNAi agent, e.g., an RNAi agent described herein inhibits the expression of the gene, mRNA, and/or protein by at least 10%, at least 20%, at least 25%, at least 30%, at least 35% or at least 40% or more, such as when assayed by a method known in the art. In some embodiments, the RNAi agent inhibits expression of a gene, mRNA, and protein by 50-100%, e.g., by 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95% and 100%.

In some embodiments, the AAV particle described herein, comprising a viral genome encoding an RNAi agent targeting a gene of interest is administered to a subject in need for treating and/or ameliorating a disease, e.g., a neurological disorder of any disease associated with the central or peripheral nervous systems.

Design of siRNA

An AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) may comprise a viral genome encoding a siRNA molecule (e.g., siRNA duplex or encoded dsRNA) that target a gene of interest and suppress target gene expression, mRNA expression, and protein production. In some aspects, the siRNA molecules are designed and used to knock out target gene variants in cells, e.g., transcripts that are identified in neurological disease. In some aspects, the siRNA molecules are designed and used to knock down target gene variants in cells.

Some guidelines for designing siRNAs (for insertion into a viral genome of the AAV particles described herein) have been proposed in the art. These guidelines generally recommend generating a 19-nucleotide duplexed region, symmetric 2-3 nucleotide 3′ overhangs, 5-phosphate and 3-hydroxyl groups targeting a region in the gene to be silenced. Other rules that may govern siRNA sequence preference include, but are not limited to, (i) A/U at the 5′ end of the antisense strand; (ii) G/C at the 5′ end of the sense strand; (iii) at least five A/U residues in the 5′ terminal one-third of the antisense strand; and (iv) the absence of any GC stretch of more than 9 nucleotides in length. In accordance with such considerations, together with the specific sequence of a target gene, highly effective siRNA molecules essential for suppressing mammalian target gene expression may be readily designed.

In one embodiment, the sense and/or antisense strand is designed based on the method and rules outlined in European Patent Publication No. EP1752536, the contents of which are herein incorporated by reference in their entirety. As a non-limiting example, the 3′-terminal base of the sequence is adenine, thymine or uracil. As a non-limiting example, the 5′-terminal base of the sequence is guanine or cytosine. As a non-limiting example, the 3′-terminal sequence comprises seven bases rich in one or more bases of adenine, thymine and uracil.

In one embodiment, an siRNA molecule comprises a sense strand and a complementary antisense strand in which both strands are hybridized together to form a duplex structure. The antisense strand has sufficient complementarity to the target mRNA sequence to direct target-specific RNAi, e.g., the siRNA molecule has a sequence sufficient to trigger the destruction of the target mRNA by the RNAi machinery or process.

In some embodiments, the antisense strand and target mRNA sequences have 100% complementarity. The antisense strand may be complementary to any part of the target mRNA sequence. Neither the identity of the sense sequence nor the homology of the antisense sequence need be 100% complementary to the target.

In other embodiments, the antisense strand and target mRNA sequences comprise at least one mismatch. As a non-limiting example, the antisense strand and the target mRNA sequence have at least 50-90%, 50-95%, 50-99%, 60-70%, 60-80%, 60-90%, 60-95%, 60-99%, 70-80%, 70-90%, 70-95%, 70-99%, 80-90%, 80-95%, 80-99%, 90-95%, 90-99% or 95-99% complementary.

The siRNA molecule may have a length from about 10-50 or more nucleotides, e.g., each strand comprising 10-50 nucleotides (or nucleotide analogs). Preferably, the siRNA molecule has a length from about 15-30, e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in each strand, wherein one of the strands is sufficiently complementary to a target region. In one embodiment, the siRNA molecule has a length from about 19 to 25, 19 to 24 or 19 to 21 nucleotides.

In some embodiments, the siRNA molecule can be a synthetic RNA duplex comprising about 19 nucleotides to about 25 nucleotides, and two overhanging nucleotides at the 3′-end.

The siRNA molecule may comprise an antisense sequence and a sense sequence, or a fragment or variant thereof. As a non-limiting example, the antisense sequence and the sense sequence have at least 50-90%, 50-95%, 50-99%, 60-70%, 60-80%, 60-90%, 60-95%, 60-99%, 70-80%, 70-90%, 70-95%, 70-99%, 80-90%, 80-95%, 80-99%, 90-95%, 90-99% or 95-99% complementary.

The sense and antisense sequences may be completely complementary across a substantial portion of their length. In other embodiments, the sense sequence and antisense sequence may be at least 70, 80, 90, 95 or 99% complementary across independently at least 50, 60, 70, 80, 85, 90, 95, or 99% of the length of the strands.

In some embodiments, the sense and antisense strands of a siRNA duplex are linked by a short spacer sequence leading to the expression of a stem-loop structure termed short hairpin RNA (shRNA). The hairpin is recognized and cleaved by Dicer, thus generating mature siRNA molecules.

In some embodiments, the siRNA molecules, as well as associated spacer and/or flanking regions once designed, can be encoded by the viral genome of the AAV particles described herein, for delivery to a cell.

Molecular Scaffold

In some embodiments, the siRNA molecules may be encoded in a modulatory polynucleotide which also comprises a molecular scaffold.

In some embodiments, the modulatory polynucleotide which comprises the payload (e.g., siRNA, miRNA or other RNAi agent described herein) includes a molecular scaffold which comprises a 5′ flanking sequence, a loop region, and/or a 3′ flanking region. In some embodiments a 5′ or 3′ flanking region may be of any length and may a wild type microRNA sequence or a portion thereof, or may be completely artificial. A 3′ flanking sequence may mirror the 5′ flanking sequence in size and origin. Either flanking sequence may be absent. In one embodiment, both the 5′ and 3′ flanking sequences are absent. The 3′ flanking sequence may optionally contain one or more CNNC motifs, where “N” represents any nucleotide. In some embodiments, the loop comprises at least one UGUG motif. In some embodiments, the UGUG motif is located at the 5′ terminus of the loop. In some embodiments the 5′ and 3′ flanking sequences are the same sequence. In some embodiments they differ by 2%, 3%, 4%, 5%, 10%, 20% or more than 30% when aligned to each other.

In some embodiments, modulatory polynucleotide comprises a stem loop structure. In some embodiments, the modulatory polynucleotide comprises in 5′ to 3′ order: a 5′ flanking sequence, a guide strand sequence, a loop region, a passenger strand sequence, and a 3′ flanking sequence. In some embodiments, the modulatory polynucleotide comprises in 5′ to 3′ order: a 5′ flanking sequence, a passenger strand sequence, a loop region, a guide strand sequence, and a 3′ flanking sequence.

In one embodiment, the molecular scaffold comprises a dual-function targeting modulatory polynucleotide.

In one embodiment, the molecular scaffold may comprise one or more linkers known in the art. The linkers may separate regions or one molecular scaffold from another. As a non-limiting example, the molecular scaffold may be polycistronic.

In one embodiment, the modulatory polynucleotide is designed using at least one of the following properties: loop variant, seed mismatch/bulge/wobble variant, stem mismatch, loop variant and basal stem mismatch variant, seed mismatch and basal stem mismatch variant, stem mismatch and basal stem mismatch variant, seed wobble and basal stem wobble variant, or a stem sequence variant.

AAV Production

Viral production disclosed herein describes processes and methods for producing AAV particles (with enhanced, improved and/or increased tropism for a target tissue), e.g., an AAV particle comprising an AAV capsid variant that may be used to contact a target cell to deliver a payload.

In some embodiments, disclosed herein is a method of making AAV particle of the present disclosure, e.g., an AAV particle comprising an AAV capsid variant the method comprising: (i) providing a host cell comprising a viral genome described herein and (ii) incubating the host cell under conditions suitable to enclose the viral genome in an AAV capsid variant, e.g., an AAV capsid variant described herein (e.g., an AAV capsid variant listed in Tables 3, 4, or 5), thereby making the AAV particle. In some embodiments, the method comprises prior to step (i), introducing a first nucleic acid comprising the viral genome into a cell. In some embodiments, the host cell comprises a second nucleic acid encoding the AAV capsid variant. In some embodiments, the second nucleic acid is introduced into the host cell prior to, concurrently with, or after the first nucleic acid molecule. In some embodiments, the AAV particle described herein is an isolated AAV particle. In some embodiments, the AAV particle described herein is a recombinant AAV particle.

Any method known in the art may be used for the preparation of AAV particles. In some embodiments, AAV particles are produced in mammalian cells (e.g., HEK293). In another embodiment, AAV particles are produced in insect cells (e.g., Sf9).

Methods of making AAV particles are well known in the art and are described in e.g., U.S. Pat. Nos. U.S. Pat. Nos. 6,204,059, 5,756,283, 6,258,595, 6,261,551, 6,270,996, 6,281,010, 6,365,394, 6,475,769, 6,482,634, 6,485,966, 6,943,019, 6,953,690, 7,022,519, 7,238,526, 7,291,498 and 7,491,508, 5,064,764, 6,194,191, 6,566,118, 8,137,948; or International Publication Nos. WO1996039530, WO1998010088, WO1999014354, WO1999015685, WO1999047691, WO2000055342, WO2000075353 and WO2001023597; Methods In Molecular Biology, ed. Richard, Humana Press, NJ (1995); O'Reilly et al., Baculovirus Expression Vectors, A Laboratory Manual, Oxford Univ. Press (1994); Samulski et al., J. Vir. 63:3822-8 (1989); Kajigaya et al., Proc. Nat'l. Acad. Sci. USA 88: 4646-50 (1991); Ruffing et al., J. Vir. 66:6922-30 (1992); Kimbauer et al., Vir., 219:37-44 (1996); Zhao et al., Vir.272:382-93 (2000); the contents of each of which are herein incorporated by reference in their entirety. In some embodiments, the AAV particles are made using the methods described in International Patent Publication WO2015191508, the contents of which are herein incorporated by reference in their entirety.

Therapeutic Applications

The present disclosure provides a method for treating a disease, disorder and/or condition in a subject, including a human subject, comprising administering to the subject an AAV particle described herein, e.g., an AAV particle comprising an AAV capsid variant (e.g., an AAV capsid variant described herein), or administering to the subject any of the described compositions, including a pharmaceutical composition, described herein.

In some embodiments, the AAV particle (e.g., an AAV particle comprising an AAV capsid variant) is administered to a subject prophylactically, to prevent on-set of disease. In another embodiment, the AAV particle (e.g., an AAV particle comprising an AAV capsid variant) is administered to treat (e.g., lessen the effects of) a disease or symptoms thereof. In yet another embodiment, the AAV particle (e.g., an AAV particle comprising an AAV capsid variant) is administered to cure (eliminate) a disease. In another embodiment, the AAV particle (e.g., an AAV particle comprising an AAV capsid variant) of the present disclosure is administered to prevent or slow progression of disease. In yet another embodiment, the AAV particle (e.g., an AAV particle comprising an AAV capsid variant) of the present disclosure are used to reverse the deleterious effects of a disease. Disease status and/or progression may be determined or monitored by standard methods known in the art.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for treatment, prophylaxis, palliation or amelioration of a genetic disorder, e.g., an autosomal dominant genetic disorder, an autosomal recessive disorder, X-linked dominant genetic disorder, an X-linked recessive genetic disorder, or a Y-linked genetic disorder. In some embodiments, the genetic disorder is a monogenetic disorder or a polygenic disorder. In some embodiments, treatment of a genetic disorder, e.g., a monogenic disorder, comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy.

In some embodiments, provided herein is method for treating a neurological disorder and/or neurodegenerative disorder in a subject, comprising administering to the subject an effective amount of a pharmaceutical composition described herein or an AAV particle, e.g., a plurality of particles, comprising an AAV capsid variant described herein. In some embodiments, treatment of a neurological disorder and/or neurodegenerative disorder comprises prevention of said neurological disorder and/or neurological disorder.

In some embodiments, the AAV particle (e.g., an AAV particle comprising an AAV capsid variant) of the disclosure is useful for the treatment, prophylaxis, palliation or amelioration of neurological diseases and/or disorders. In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of tauopathy.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is for the treatment, prophylaxis, palliation or amelioration of Alzheimer's Disease. In some embodiments, treatment of Alzheimer's Disease comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an ApoE2 protein, ApoE4 protein, an ApoE3 protein, BDNF protein, CYP46A1 protein, Klotho protein, fractalkine (FKN) protein, neprilysin protein (NEP), CD74 protein, caveolin-1, or a combination or variant thereof. In some embodiments, treatment of Alzheimer's Disease comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a reduction in the expression of a tau gene and/or protein, a synuclein gene and/or protein, or a combination or variant thereof. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an antibody that binds to tau or synuclein, an RNAi agent for inhibiting tau or synuclein, a gene editing system (e.g., a CRISPR-Cas system) for altering tau or synuclein expression, or a combination thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful the treatment, prophylaxis, palliation or amelioration of Friedreich's ataxia, or any disease stemming from a loss or partial loss of frataxin protein.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is for the treatment, prophylaxis, palliation or amelioration of frontal temporal dementia. In some embodiments, treatment of frontal temporal dementia comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a progranulin protein or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of Parkinson's Disease. In some embodiments, treatment of Parkinson's disease comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an AADC protein, GAD protein, GDNF protein, TH-GCH1 protein, GBA protein, AIMP2-DX2 protein, or a combination or variant thereof. In some embodiments, treatment of Parkinson's disease comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene knock-down therapy or a gene editing therapy (e.g., knock-out, repression, or correction). In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a modulator, e.g., an RNAi agent or a CRISPR-Cas system, for altering expression of an alpha-synuclein gene, mRNA, and/or protein, or variant thereof. In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of an AADC deficiency. In some embodiments, treatment of AADC deficiency comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an AADC protein or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of Amyotrophic lateral sclerosis. In some embodiments, treatment of ALS comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a TDP-43 protein, UPF1 protein, C9orf72 protein, CCNF protein, HSF1 protein, Factor H protein, NGF protein, ADAR2 protein, GDNF protein, VEGF protein, HGF protein, NRTN protein, AIMP2-DX2 protein, or a combination or variant thereof. In some embodiments, treatment of ALS comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene knock-down therapy or a gene editing therapy (e.g., knock-out, repression, or correction). In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a modulator, e.g., an RNAi agent or a CRISPR-Cas system, for altering expression of a SOD1 or C90RF72 gene, mRNA, and/or protein, or a combination or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of Huntington's Disease. In some embodiments, treatment of ALS comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene knock-down (e.g., knock-out) therapy or a gene editing therapy (e.g., knock-out, repression, or correction). In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a modulator, e.g., an RNAi agent or a CRISPR-Cas system, for altering expression of an HTT gene, mRNA, and/or protein, or a variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of spinal muscular atrophy. In some embodiments, treatment of spinal muscular atrophy comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an SMN1 protein, an SMN2 protein, or a combination or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of multiple system atrophy. In some embodiments, treatment of multiple system atrophy comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of Gaucher disease (GD) (e.g., Type 1 GD, Type 2 GD, or Type 3 GD). In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of Parkinson's disease associated with a GBA mutation. In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of dementia with Lewy Bodies (DLB).

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for treatment, prophylaxis, palliation or amelioration of a leukodystrophy, e.g., Alexander disease, autosomal dominant leukodystrophy with autonomic diseases (ADLD), Canavan disease, cerebrotendinous xanthomatosis (CTX), metachromatic leukodystrophy (MLD), Pelizaeus-Merzbacher disease, or Refsum disease. In some embodiments, treatment of MLD comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an ARSA protein or variant thereof. In some embodiments, treatment of ALD comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an ABCD-1 protein or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of megalencephalic leukoencephalopathy (MLC). In some embodiments, treatment of MLC comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an MLC1 protein or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of Krabbe disease. In some embodiments, treatment of Krabbe disease comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a GALC protein or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of Mucopolysaccharidosis, e.g., a Type I (MPS I), Type II (MPS II), Type IIIA (MPS IIIA), Type IIIB (MPS IIIB), or Type IIIC (MPS IIIC). In some embodiments, treatment of Mucopolysaccharidosis comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy or a gene editing therapy (e.g., enhancement or correction). In some embodiments, the payload encoded or corrected by an AAV particle comprising a capsid variant described herein comprises an IDUA protein, IDS protein, SGSH protein, NAGLU protein, HGSNAT protein, or a combination or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of Batten/NCL. In some embodiments, treatment of Batten/NCL comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a CLN1 protein, CLN2 protein, CLN3 protein, CLN5 protein, CLN6 protein, CLN7 protein, CLN8 protein, or a combination or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of Rett Syndrome. In some embodiments, treatment of Rett Syndrome comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an MeCP2 protein or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of Angelman Syndrome. In some embodiments, treatment of Angelman Syndrome comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a UBE3A protein or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of Fragile X Syndrome. In some embodiments, treatment of Fragile X Syndrome comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a Reelin protein, a DgkK protein, a FMR1 protein, or a combination or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of Canavan Disease. In some embodiments, treatment of Canavan Disease comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an ASPA protein or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of a Gangliosidosis, e.g., a GM1 Gangliosidosis or a GM2 Gangliosidosis (e.g., Tay Sachs Sandhoff). In some embodiments, treatment of a Gangliosidosis, e.g., a GM1 Gangliosidosis or a GM2 Gangliosidosis (e.g., Tay Sachs Sandhoff), comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a GLB1 protein, a HEXA protein, a HEXB protein, a GM2A protein, or a combination or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of GM3 Synthase Deficiency. In some embodiments, treatment of GM3 Synthase Deficiency comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an ST3GAL5 protein or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of a Niemann-Pick disorder, e.g., a Niemann-Pick A or a Niemann-Pick C1 (NPC-1). In some embodiments, treatment of a Niemann-Pick disorder, e.g., a Niemann-Pick A or a Niemann-Pick C1 (NPC-1) comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an ASM protein, an NPC1 protein, or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of Schwannoma (e.g., Neuroma). In some embodiments, treatment of Schwannoma (e.g., Neuroma) comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a Caspase-1 protein or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of a Tuberous Sclerosis, e.g., Tuberous Sclerosis Type 1 or Tuberous Sclerosis Type 2. In some embodiments, treatment of Tuberous Sclerosis, e.g., Tuberous Sclerosis Type 1 or Tuberous Sclerosis Type 2 comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a TSC1 protein, a TSC2 protein, or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of a CDKL5 Deficiency. In some embodiments, treatment of a CDKL5 Deficiency comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a CDKL5 protein or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of a Charcot-Marie-Tooth disorder, e.g., a Charcot-Marie-Tooth Type 1× (CMT1X) disorder, a Charcot-Marie-Tooth Type 2A (CMT2A) disorder, or a Charcot-Marie-Tooth Type 4J (CMT4J) disorder. In some embodiments, treatment of a Charcot-Marie-Tooth disorder, e.g., a Charcot-Marie-Tooth Type 1X (CMT1X) disorder, a Charcot-Marie-Tooth Type 2A (CMT2A) disorder, or a Charcot-Marie-Tooth Type 4J (CMT4J) disorder, comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a GJB1 protein, a MFN2 protein, a FIG4 protein, or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of an Aspartylglucosaminuria (AGU). In some embodiments, treatment of an AGU comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an AGA protein or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of a Leigh Syndrome. In some embodiments, treatment of a Leigh Syndrome comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a SURF1 protein or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of epilepsy. In some embodiments, treatment of epilepsy comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an NPY/Y2 protein, a Galanin protein, a Dynorphin protein, an AIMP2-DX2 protein, an SLC6A1 protein, an SLC13A5 protein, a KCNQ2 protein, or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of a Dravet Syndrome. In some embodiments, treatment of Dravet Syndrome comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises an SCN1a protein, or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of a Duchenne muscular dystrophy (DMD). In some embodiments, treatment of DMD comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy or enhancement (e.g., correction of exon-skipping), or a gene editing therapy (e.g., enhancement or correction). In some embodiments, the payload encoded or corrected by an AAV particle comprising a capsid variant described herein comprises a Dystrophin gene and/or protein, a Utrophin gene and/or protein, or a GALGT2 gene and/or protein, or a Follistatin gene and/or protein, or a combination or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of Pompe Disease. In some embodiments, treatment of Pompe Disease comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a GAA protein, or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation, or amelioration of Limb-Girdle Muscular Dystrophy (LGMD2A). In some embodiments, treatment of LGMD2A comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy. In some embodiments, the payload encoded by an AAV particle comprising a capsid variant described herein comprises a CAPN-3 protein, DYSF protein, a SGCG protein, a SGCA protein, a SGCB protein, a FKRP protein, a ANO5 protein, or a combination or variant thereof.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of chronic or neuropathic pain.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising AAV capsid variant) is useful for treatment, prophylaxis, palliation or amelioration of a disease associated with the central nervous system.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for treatment, prophylaxis, palliation or amelioration of a disease associated with the peripheral nervous system.

In some embodiments, provided herein is a method for treating a neuro-oncological disorder in a subject, comprising administering to the subject an effective amount of a pharmaceutical composition described herein or an AAV particle, e.g., a plurality of particles, comprising an AAV capsid variant described herein. In some embodiments, treatment of a neuro-oncological disorder comprises prevention of said neuro-oncological disorder. In some embodiments, a neuro-oncological disorder comprises a cancer of a primary CNS origin (e.g., a CNS cell, a tissue, or a region), or a metastatic cancer in a CNS cell, tissue, or region. Examples of primary CNS cancers could be gliomas (which may include glioblastoma (also known as glioblastoma multiforme), astrocytomas, oligodendrogliomas, and ependymomas, and mixed gliomas), meningiomas, medulloblastomas, neuromas, and primary CNS lymphoma (in the brain, spinal cord, or meninges), among others. Examples of metastatic cancers include those originating in another tissue or organ, e.g., breast, lung, lymphoma, leukemia, melanoma (skin cancer), colon, kidney, prostate, or other types that metastasize to brain.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of a disease associated with expression of HER2, e.g., a disease associated with overexpression of HER2. In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of a HER2-positive cancer. In some embodiments, the HER2-positive cancer is a HER2-positive solid tumor. Additionally, or alternatively, the HER2-positive cancer may be a locally advanced or metastatic HER2-positive cancer. In some instances, the HER2-positive cancer is a HER2-positive breast cancer or a HER2-positive gastric cancer. In some embodiments, the HER2-positive cancer is selected from the group consisting of a HER2-positive gastroesophageal junction cancer, a HER2-positive colorectal cancer, a HER2-positive lung cancer (e.g., a HER2-positive non-small cell lung carcinoma), a HER2-positive pancreatic cancer, a HER2-positive colorectal cancer, a HER2-positive bladder cancer, a HER2-positive salivary duct cancer, a HER2-positive ovarian cancer (e.g., a HER2-positive epithelial ovarian cancer), or a HER2-positive endometrial cancer. In some instances, the HER2-positive cancer is prostate cancer. In some embodiments, the HER2-positive cancer has metastasized to the central nervous system (CNS). In some instances, the metastasized HER2-cancer has formed CNS neoplasms.

In some embodiments, the AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant) is administered to a subject having at least one of the diseases or symptoms described herein. In some embodiments, an AAV particle of the present disclosure is administered to a subject having or diagnosed with having a disease or disorder described herein.

In some embodiments, provided herein is a method for treating a muscular disorder and/or neuromuscular disorder in a subject, comprising administering to the subject an effective amount of a pharmaceutical composition described herein or an AAV particle, e.g., a plurality of particles, comprising an AAV capsid variant described herein. In some embodiments, treatment of a muscular disorder and/or neuromuscular disorder comprises prevention of said muscular disorder and/or neuromuscular disorder.

In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for treatment, prophylaxis, palliation or amelioration of a cardiac disease or heart disease and/or method of improving (e.g., enhancing) cardiac function in a subject. In some embodiments, the cardiac disease is a cardiomyopathy (e.g., arrhythmogenic right ventricular cardiomyopathy, dilated cardiomyopathy, or hypertrophic cardiomyopathy), congestive heart failure, tachycardia (e.g., catecholaminergic polymorphic ventricular tachycardia), ischemic heart disease, and/or myocardial infarction. In some embodiments, the cardiac disease is a disease associated with expression, e.g., aberrant expression, of LAMP2B, MYBPC3, TNNI3, LMNA, BAG3, DWORF, PKP2, Cx43, TAZ, CASQ2, SERCA2a, I-1c, S100A1 and/or ARC, S100A1, ASCL1, miR133, Mydelta3, Sav, or a combination or variant thereof. In some embodiments, treatment of a cardiac disorder described herein comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy.

In some embodiments, the cardiac disease is a genetic disorder, e.g., an autosomal dominant genetic disorder, an autosomal recessive disorder, or an X-linked recessive genetic disorder. In some embodiments, the cardiomyopathy is a genetic disorder, e.g., a genetic disorder associated with an abnormality (e.g., mutation, insertion, rearrangement and/or deletion) in a gene chosen from TTN, LMNA, MYH7, MYH6, SCN5A, TNNT2, RBM20, TNNI3, MYL2, MYL3, PKP2, DSP, DSG2, DSC2, JUP, or a combination thereof. In some embodiments, the cardiac disorder is a dilated cardiomyopathy, e.g., a dilated cardiomyopathy associated with an abnormality (e.g., mutation, insertion, rearrangement and/or deletion) in a gene chosen from TTN, LMNA, MIH7, BAG3, MIPN, TNNT2, SCN5A, RBN20, TNPO, LAMA4, VCL, LDB3, TCAP, PSEN1/2, ACTN2, CRYAB, TPM1, ABCC9, ACTC1, PDLIM3, ILK, TNNC1, TNNI3, PLN, DES, SGCD, CSRP3, MIH6, EYA4, ANKRD1, DMD, GATADI, TAZ/G4.5, or combination thereof. In some embodiments, the cardiac disorder is a hypertrophic cardiomyopathy, e.g., a hypertrophic cardiomyopathy associated with an abnormality (e.g., mutation, insertion, rearrangement and/or deletion) in a gene chosen from MYH7, TNNT2, TNNI3, TPM1, MYL2, MYL3, ACTC1, CSRP3, TTN, ACTN2, MYH6, TCAP, TNNC1, or a combination thereof. In some embodiments, the cardiac disorder is an arrhythmogenic ventricular cardiomyopathy, e.g., an arrhythmogenic ventricular cardiomyopathy associated with an abnormality (e.g., mutation, insertion, rearrangement and/or deletion) in a gene chosen from PKP2, DSG2, DSP, RYR2, DSC2, TGFB3, TMEM43, DES, TTN, LMNA, or a combination thereof.

In some embodiments, the AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant) is administered to a subject having at least one of the diseases or symptoms described herein. In some embodiments, an AAV particle of the present disclosure is administered to a subject having or diagnosed with having a disease or disorder described herein.

Any neurological disease or disorder, neurodegenerative disorder, muscular disorder, neuromuscular disorder, and/or neuro-oncological disorder may be treated with the AAV particles of the disclosure, or pharmaceutical compositions thereof.

Pharmaceutical Composition and Formulations

According to the present disclosure, an AAV particle comprising an AAV capsid variant described herein may be prepared as a pharmaceutical composition. In some embodiments, the pharmaceutical composition comprises at least one active ingredients. In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable excipient.

In some embodiments, an AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant) can be formulated using an excipient to: (1) increase stability; (2) increase cell transfection or transduction; (3) permit the sustained or delayed expression of the payload; (4) alter the biodistribution (e.g., target the viral particle to specific tissues or cell types); (5) increase the translation of encoded protein; (6) alter the release profile of encoded protein; and/or (7) allow for regulatable expression of the payload. Formulations of the present disclosure can include, without limitation, saline, liposomes, lipid nanoparticles, polymers, peptides, proteins, cells transfected with viral vectors (e.g., for transfer or transplantation into a subject) and combinations thereof.

In some embodiments, the relative amount of the active ingredient (e.g. an AAV particle comprising an AAV capsid variant described herein), a pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition in accordance with the present disclosure may vary, depending upon the identity, size, and/or condition of the subject being treated and further depending upon the route by which the composition is to be administered. For example, the composition may comprise between 0.1% and 99% (w/w) of the active ingredient. By way of example, the composition may comprise between 0.1% and 100%, e.g., between 0.5 and 50%, between 1-30%, between 5-80%, at least 80% (w/w) active ingredient.

In some embodiments, the pharmaceutical composition comprising an AAV particle described herein may comprise an AAV capsid variant and a viral genome encoding a payload, e.g., a payload described herein, with or without a pharmaceutically acceptable excipient.

The present disclosure also provides in some embodiments, a pharmaceutical composition suitable for administration to a subject, e.g., a human. In some embodiments, the pharmaceutical composition is administered to a subject, e.g., a human.

Administration

In some embodiments, an AAV particle disclosed herein (e.g., an AAV particle comprising an AAV capsid variant) may be administered to a subject by a delivery route, e.g., a localized delivery route or a systemic delivery route.

In some embodiments, an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant) may be administered via such a route that it is able to cross the blood-brain barrier, vascular barrier, or other epithelial barrier. In some embodiments, an AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) may be administered in any suitable form, either as a liquid solution or suspension, as a solid form suitable for liquid solution or suspension in a liquid solution. In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant) may be formulated with any appropriate and pharmaceutically acceptable excipient.

In some embodiments, the AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant) is administered intramuscularly, intravenously, intracerebrally, intrathecally, intratumorally, intracerebroventricularly, via intraparenchymal administration, or via intra-cisterna magna injection (ICM). In some embodiments, the AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant) is administered intravenously. In some embodiments, the AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant) is administered via intra-cisterna magna injection (ICM). In some embodiments, the AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant) is administered intratumorally. In some embodiments, the AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant) is administered intraarterially.

In some embodiments, an AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) may be delivered to a subject via a single route administration. In some embodiments, an AAV particle of the present disclosure may be delivered to a subject via a multi-site route of administration. In some embodiments, a subject may be administered at 2, 3, 4, 5, or more than 5 sites.

In some embodiments, an AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) is administered via a bolus infusion. In some embodiments, an AAV particle of the present disclosure is administered via sustained delivery over a period of minutes, hours, or days. In some embodiments, the infusion rate may be changed depending on the subject, distribution, formulation, and/or another delivery parameter. In some embodiments, an AAV particle of the present disclosure is administered using a controlled release. In some embodiments, an AAV particle of the present disclosure is administered using a sustained release, e.g., a release profile that conforms to a release rate over a specific period of time.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant) may be delivered by more than one route of administration. As non-limiting examples of combination administrations, an AAV particle may be delivered by intrathecal and intracerebroventricular, or by intravenous and intraparenchymal administration.

Intravenous Administration

In some embodiments, an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant) may be administered to a subject by systemic administration. In some embodiments, the systemic administration is intravenous administration. In another embodiment, the systemic administration is intraarterial administration. In some embodiments, an AAV particle of the present disclosure may be administered to a subject by intravenous administration. In some embodiments, the intravenous administration may be achieved by subcutaneous delivery. In some embodiments, the AAV particle is administered to the subject via focused ultrasound (FUS), e.g., coupled with the intravenous administration of microbubbles (FUS-MB) or MRI-guided FUS coupled with intravenous administration, e.g., as described in Terstappen et al. (Nat Rev Drug Discovery, doi.org/10.1038/s41573-021-00139-y (2021)), the contents of which are incorporated herein by reference in its entirety. In some embodiments, the AAV particle is administered to the subject intravenously. In some embodiments, the subject is a human.

Administration to the CNS

In some embodiments, an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant) may be delivered by direct injection into the brain. As a non-limiting example, the brain delivery may be by intrahippocampal administration. In some embodiments, an AAV particle of the present disclosure may be administered to a subject by intraparenchymal administration. In some embodiments, the intraparenchymal administration is to tissue of the central nervous system. In some embodiments, an AAV particle of the present disclosure may be administered to a subject by intracranial delivery (See, e.g., U.S. Pat. No. 8,119,611; the content of which is incorporated herein by reference in its entirety). In some embodiments, an AAV particle described herein may be delivered by injection into the CSF pathway. Non-limiting examples of delivery to the CSF pathway include intrathecal and intracerebroventricular administration. In some embodiments, an AAV particle described herein may be administered via intracisternal magna (ICM) injection.

In some embodiments, an AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) may be delivered to the brain by systemic delivery. As a non-limiting example, the systemic delivery may be by intravascular administration. As a non-limiting example, the systemic or intravascular administration may be intravenous.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant) of the present disclosure may be delivered by an intraocular delivery route. A non-limiting example of an intraocular administration includes an intravitreal injection.

Intramuscular Administration

In some embodiments, an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant) may be delivered by intramuscular administration. Without wishing to be bound by theory, it is believed in some embodiments, that the multi-nucleated nature of muscle cells provides an advantage to gene transduction subsequent to AAV delivery. In some embodiments, cells of the muscle are capable of expressing recombinant proteins with the appropriate post-translational modifications. Without wishing to be bound by theory, it is believed in some embodiments, the enrichment of muscle tissue with vascular structures allows for transfer to the blood stream and whole-body delivery. Examples of intramuscular administration include systemic (e.g., intravenous), subcutaneous or directly into the muscle. In some embodiments, more than one injection is administered. In some embodiments, an AAV particle of the present disclosure may be delivered by an intramuscular delivery route. (See, e.g., U.S. Pat. No. 6,506,379; the content of which is incorporated herein by reference in its entirety). Non-limiting examples of intramuscular administration include an intravenous injection or a subcutaneous injection.

In some embodiments, an AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) is administered to a subject and transduces the muscle of a subject. As a non-limiting example, an AAV particle is administered by intramuscular administration. In some embodiments, an AAV particle of the present disclosure may be administered to a subject by subcutaneous administration. In some embodiments, the intramuscular administration is via systemic delivery. In some embodiments, the intramuscular administration is via intravenous delivery. In some embodiments, the intramuscular administration is via direct injection to the muscle.

In some embodiments, the muscle is transduced by administration, e.g., intramuscular administration. In some embodiments, an intramuscular delivery comprises administration at one site. In some embodiments, an intramuscular delivery comprises administration at more than one site. In some embodiments, an intramuscular delivery comprises administration at two, three, four, or more sites. In some embodiments, intramuscular delivery is combined with at least one other method of administration.

In some embodiments, an AAV particle pf the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) may be administered to a subject by peripheral injections. Non-limiting examples of peripheral injections include intraperitoneal, intramuscular, intravenous, conjunctival, or joint injection. It was disclosed in the art that the peripheral administration of AAV vectors can be transported to the central nervous system, for example, to the motor neurons (e.g., U. S. Patent Publication Nos. US20100240739 and US20100130594; the content of each of which is incorporated herein by reference in their entirety).

In some embodiments, an AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) may be administered to a subject by intraparenchymal administration. In some embodiments, the intraparenchymal administration is to muscle tissue. In some embodiments, an AAV particle of the present disclosure is delivered as described in Bright et al 2015 (Neurobiol Aging. 36(2):693-709), the contents of which are herein incorporated by reference in their entirety. In some embodiments, an AAV particle of the present disclosure is administered to the gastrocnemius muscle of a subject. In some embodiments, an AAV particle of the present disclosure is administered to the bicep femorii of the subject. In some embodiments, an AAV particles of the present disclosure is administered to the tibialis anterior muscles. In some embodiments, an AAV particle of the present disclosure is administered to the soleus muscle.

Depot Administration

In some embodiments, a pharmaceutical composition and/or an AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) are formulated in depots for extended release. Generally, specific organs or tissues are targeted for administration.

In some embodiments, a pharmaceutical composition and/or an AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) are spatially retained within or proximal to target tissues. Provided are methods of providing a pharmaceutical composition, an AAV particle, to target tissues of mammalian subjects by contacting target tissues (which comprise one or more target cells) with the pharmaceutical composition and/or the AAV particle, under conditions such that they are substantially retained in target tissues, e.g., such that at least 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, 96, 97, 98, 99, 99.9, 99.99 or greater than 99.99% of the composition is retained in the target tissues. In some embodiments, retention is determined by measuring the amount of pharmaceutical composition and/or AAV particle, that enter a target cell or a plurality of target cells. For example, at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.9%, 99.99%, or greater than 99.99% of a pharmaceutical composition and/or an AAV particle, administered to a subject are present intracellularly at a period of time following administration. For example, intramuscular injection to a subject may be performed using aqueous compositions comprising a pharmaceutical composition and/or an AAV particle of the present disclosure and a transfection reagent, and retention is determined by measuring the amount of the pharmaceutical composition and/or the AAV particle, present in the muscle cell or plurality of muscle cells.

In some embodiments, disclosed herein are methods of providing a pharmaceutical composition and/or an AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) to a tissue of a subject, by contacting the tissue (comprising a cell, e.g., a plurality of cells) with the pharmaceutical composition and/or the AAV particle under conditions such that they are substantially retained in the tissue. In some embodiments, a pharmaceutical composition and/or AAV particle described herein comprise a sufficient amount of an active ingredient such that the effect of interest is produced in at least one cell. In some embodiments, a pharmaceutical composition and/or an AAV particle generally comprise one or more cell penetration agents. In some embodiments, the disclosure provides a naked formulations (such as without cell penetration agents or other agents), with or without pharmaceutically acceptable carriers.

Methods of Treatment

Provided in the present disclosure are methods for introducing (e.g., delivering) an AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) into cells. In some embodiments, the method comprises introducing into said cells an AAV particle or vector described herein in an amount sufficient to modulate, e.g., increase, the production of a target gene, mRNA, and/or protein. In some embodiments, the method comprises introducing into said cells an AAV particle or vector described herein in an amount sufficient to modulate, e.g., decrease, expression of a target gene, mRNA, and/or protein. In some aspects, the cells may be neurons such as but not limited to, motor, hippocampal, entorhinal, thalamic, cortical, sensory, sympathetic, or parasympathetic neurons, and glial cells such as astrocytes, microglia, and/or oligodendrocytes. In other aspects, the cells may be a muscle cell, e.g., a cell of a diaphragm, a quadriceps, or a heart (e.g., a heart atrium or a heart ventricle). In other embodiments, the cells may be a muscle cell (e.g., a cell of a diaphragm, a quadriceps, or a heart (e.g., a heart atrium or a heart ventricle)) or a liver cell. In some embodiments, the cell may be a heart cell (e.g., a cell of a heart atrium or a cell of a heart ventricle).

Disclosed in the present disclosure are methods for treating a neurological disease/disorder or a neurodegenerative disorder, a muscular or neuromuscular disorder, or a neurooncological disorder associated with aberrant, e.g., insufficient or increased, function/presence of a protein, e.g., a target protein in a subject in need of treatment.

In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a composition comprising AAV particles of the present disclosure. As a non-limiting example, the AAV particles can increase target gene expression, increase target protein production, and thus reduce one or more symptoms of neurological disease in the subject such that the subject is therapeutically treated.

In other embodiments, the method comprises administering to the subject a therapeutically effective amount of a composition comprising AAV particles (e.g., an AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant) comprising a viral genome with a nucleic acid sequence encoding one or more siRNA molecules. As a non-limiting example, the siRNA molecules can silence target gene expression, inhibit target protein production, and reduce one or more symptoms of neurological disease in the subject such that the subject is therapeutically treated.

In some embodiments, the composition comprising the AAV particles of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant described herein) is administered to the central nervous system of the subject via systemic administration. In some embodiments, the systemic administration is intravenous (IV) injection. In some embodiments, the AAV particle described herein or a pharmaceutical composition comprising an AAV particle described herein is administered by focused ultrasound (FUS), e.g., coupled with the intravenous administration of microbubbles (FUS-MB) or MRI-guided FUS coupled with intravenous administration.

In some embodiments, the composition comprising the AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) is administered to the central nervous system of the subject via intraventricular administration. In some embodiments, the composition comprising the AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) is administered via intra-cisterna magna injection (ICM).

In some embodiments, the composition comprising an AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) is administered to the central nervous system of the subject via intraventricular injection and intravenous injection.

In some embodiments, the composition comprising the AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) is administered to the central nervous system of the subject via ICM injection and intravenous injection at a specific dose per subject. As a non-limiting example, the AAV particles are administered via ICM injection at a dose of 1×10⁴VG per subject. As a non-limiting example, the AAV particles are administered via IV injection at a dose of 2×10¹³VG per subject.

In some embodiments, the composition comprising the AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) is administered to the central nervous system of the subject. In other embodiments, the composition comprising the AAV particles of the present disclosure is administered to a CNS tissue of a subject (e.g., putamen, hippocampus, thalamus, or cortex of the subject).

In some embodiments, the composition comprising the AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) is administered to the central nervous system of the subject via intraparenchymal injection. Non-limiting examples of intraparenchymal injections include intraputamenal, intracortical, intrathalamic, intrastriatal, intrahippocampal or into the entorhinal cortex.

In some embodiments, the composition comprising the AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) is administered to the central nervous system of the subject via intraparenchymal injection and intravenous injection.

In some embodiments, the composition comprising the AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) is administered to the central nervous system of the subject via intraventricular injection, intraparenchymal injection and intravenous injection.

In some embodiments, the composition comprising an AAV particle (e.g., an AAV particle comprising an AAV capsid variant) of a plurality of particles of the present disclosure is administered to a muscle of the subject via intravenous injection. In some embodiments, the composition comprising an AAV particle of a plurality of particles of the present disclosure is administered to a muscle of the subject via intramuscular injection.

In some embodiments, an AAV particle of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) may be delivered into specific types of cells, including, but not limited to, thalamic, hippocampal, entorhinal, cortical, motor, sensory, excitatory, inhibitory, sympathetic, or parasympathetic neurons; glial cells including oligodendrocytes, astrocytes and microglia; and/or other cells surrounding neurons such as T cells. In some embodiments, an AAV particle of the present disclosure may be delivered into a muscle cell, e.g., a cell of the quadriceps, diaphragm, liver, and/or heart (e.g., heart atrium or heart ventricle).

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be delivered to a cell or region of the midbrain. In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be delivered to a cell or region of the brains stem.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be delivered to neurons in the putamen, hippocampus, thalamus and/or cortex.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for a genetic disorder, e.g., an autosomal dominant genetic disorder, an autosomal recessive disorder, X-linked dominant genetic disorder, an X-linked recessive genetic disorder, or a Y-linked genetic disorder. In some embodiments, the genetic disorder is a monogenetic disorder or a polygenic disorder. In some embodiments, treatment of a genetic disorder, e.g., a monogenic disorder, comprises the use of an AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant described herein) for a gene replacement therapy.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for a neurological disease.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for tauopathies.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for Alzheimer's Disease.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for Amyotrophic Lateral Sclerosis.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for Huntington's Disease.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for Parkinson's Disease.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for Gaucher disease (GD) (e.g., Type 1 GD, Type 2 GD, or Type 3 GD). In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for Parkinson's disease associated with a GBA mutation. In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for dementia with Lewy Bodies (DLB).

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for spinal muscular atrophy.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for a leukodystrophy, e.g., Alexander disease, autosomal dominant leukodystrophy with autonomic diseases (ADLD), Canavan disease, cerebrotendinous xanthomatosis (CTX), metachromatic leukodystrophy (MLD), Pelizaeus-Merzbacher disease, or Refsum disease.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for Friedreich's Ataxia.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for chronic or neuropathic pain.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for a muscular disorder or a neuromuscular disorder.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for a cardiac disease or heart disease and/or method of improving (e.g., enhancing) cardiac function in a subject. In some embodiments, the cardiac disease is a cardiomyopathy (e.g., arrhythmogenic right ventricular cardiomyopathy, dilated cardiomyopathy, or hypertrophic cardiomyopathy), congestive heart failure, tachycardia (e.g., catecholaminergic polymorphic ventricular tachycardia), ischemic heart disease, and/or myocardial infarction.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant), e.g., a plurality of particles, of the present disclosure may be used as a therapy for a disease associated with expression of HER2, e.g., a disease associated with overexpression of HER2. In some embodiments, the AAV particle of the disclosure (e.g., an AAV particle comprising an AAV capsid variant) is useful for the treatment, prophylaxis, palliation or amelioration of a HER2-positive cancer. In some embodiments, the HER2-positive cancer is a HER2-positive solid tumor. Additionally, or alternatively, the HER2-positive cancer may be a locally advanced or metastatic HER2-positive cancer. In some instances, the HER2-positive cancer is a HER2-positive breast cancer or a HER2-positive gastric cancer. In some embodiments, the HER2-positive cancer is selected from the group consisting of a HER2-positive gastroesophageal junction cancer, a HER2-positive colorectal cancer, a HER2-positive lung cancer (e.g., a HER2-positive non-small cell lung carcinoma), a HER2-positive pancreatic cancer, a HER2-positive colorectal cancer, a HER2-positive bladder cancer, a HER2-positive salivary duct cancer, a HER2-positive ovarian cancer (e.g., a HER2-positive epithelial ovarian cancer), or a HER2-positive endometrial cancer. In some instances, the HER2-positive cancer is prostate cancer. In some embodiments, the HER2-positive cancer has metastasized to the central nervous system (CNS). In some instances, the metastasized HER2-cancer has formed CNS neoplasms.

In some embodiments, an AAV particle (e.g., an AAV particle comprising an AAV capsid variant) e.g., a plurality of particles, of the present disclosure may be used as a therapy for a neuro-oncological disorder. In some embodiments, the neuro-oncological disorder is a cancer of primary CNS origin (e.g., a cancer of a CNS cell and/or CNS tissue). In some embodiments, the neuro-oncological disorder is metastatic cancer in a CNS cell, CNS region, and/or a CNS tissue. Examples of primary CNS cancers could be gliomas (which may include glioblastoma (also known as glioblastoma multiforme), astrocytomas, oligodendrogliomas, and ependymomas, and mixed gliomas), meningiomas, medulloblastomas, neuromas, and primary CNS lymphoma (in the brain, spinal cord, or meninges), among others. Examples of metastatic cancers include those originating in another tissue or organ, e.g., breast, lung, lymphoma, leukemia, melanoma (skin cancer), colon, kidney, prostate, or other types that metastasize to brain.

In some embodiments, administration of the AAV particle described herein (e.g., an AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant) to a subject may increase target gene, mRNA, and/or protein levels in a subject, relative to a control, e.g., the gene, mRNA, and/or mRNA levels in the subject prior to receiving AAV particle. The target gene, mRNA, and/or protein levels may be increased by about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95% and 100%, or at least 20-30%, 20-40%, 20-50%, 20-60%, 20-70%, 20-80%, 20-90%, 20-95%, 20-100%, 30-40%, 30-50%, 30-60%, 30-70%, 30-80%, 30-90%, 30-95%, 30-100%, 40-50%, 40-60%, 40-70%, 40-80%, 40-90%, 40-95%, 40-100%, 50-60%, 50-70%, 50-80%, 50-90%, 50-95%, 50-100%, 60-70%, 60-80%, 60-90%, 60-95%, 60-100%, 70-80%, 70-90%, 70-95%, 70-100%, 80-90%, 80-95%, 80-100%, 90-95%, 90-100% or 95-100% in a subject such as, but not limited to, the CNS, a region of the CNS, or a specific cell of the CNS, or a muscle, a region of a muscle, or a cell of a muscle, of a subject. In some embodiments, cell of the CNS comprises an astrocyte, microglia, cortical neuron, hippocampal neuron, DRG and/or sympathetic neuron, sensory neuron, oligodendrocyte, motor neuron, or combination thereof. As a non-limiting example, the AAV particles may increase the gene, mRNA, and/or protein levels of a target protein by fold increases over baseline. In some embodiments, AAV particles lead to 5-6 times higher levels of a target gene, mRNA, or protein.

In some embodiments, administration of the AAV particle described herein (e.g., an AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant), e.g., an AAV particle comprising a nucleic acid encoding a siRNA molecule, to a subject may decrease target gene, mRNA, and/or protein levels in a subject, relative to a control, e.g., the gene, mRNA, and/or mRNA levels in the subject prior to receiving AAV particle. The target gene, mRNA, and/or protein levels may be decreased by about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95% and 100%, or at least 20-30%, 20-40%, 20-50%, 20-60%, 20-70%, 20-80%, 20-90%, 20-95%, 20-100%, 30-40%, 30-50%, 30-60%, 30-70%, 30-80%, 30-90%, 30-95%, 30-100%, 40-50%, 40-60%, 40-70%, 40-80%, 40-90%, 40-95%, 40-100%, 50-60%, 50-70%, 50-80%, 50-90%, 50-95%, 50-100%, 60-70%, 60-80%, 60-90%, 60-95%, 60-100%, 70-80%, 70-90%, 70-95%, 70-100%, 80-90%, 80-95%, 80-100%, 90-95%, 90-100% or 95-100% in a subject such as, but not limited to, the CNS, a region of the CNS, or a specific cell of the CNS, or a muscle, a region of a muscle, or a cell of a muscle, of a subject. In some embodiments, cell of the CNS comprises an astrocyte, microglia, cortical neuron, hippocampal neuron, DRG and/or sympathetic neuron, sensory neuron, oligodendrocyte, motor neuron, or combination thereof. As a non-limiting example, the AAV particles may decrease the gene, mRNA, and/or protein levels of a target protein by fold decreases over baseline.

In some embodiments, the AAV particles of the present disclosure (e.g., an AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant) may be used to increase target protein and reduce symptoms of neurological disease in a subject. In some embodiments, the AAV particles of the present disclosure (e.g., an AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant) may be used to decrease target protein and reduce symptoms of neurological disease in a subject.

In some embodiments, the AAV particles of the present disclosure (e.g., an AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant) may be used to reduce the decline of functional capacity and activities of daily living as measured by a standard evaluation system such as, but not limited to, the total functional capacity (TFC) scale.

In some embodiments, the AAV particles of the present disclosure (e.g., an AAV particle comprising an AAV capsid polypeptide, e.g., an AAV capsid variant) may be used to improve performance on any assessment used to measure symptoms of neurological disease. Such assessments include, but are not limited to ADAS-cog (Alzheimer Disease Assessment Scale-cognitive), MMSE (Mini-Mental State Examination), GDS (Geriatric Depression Scale), FAQ (Functional Activities Questionnaire), ADL (Activities of Daily Living), GPCOG (General Practitioner Assessment of Cognition), Mini-Cog, AMTS (Abbreviated Mental Test Score), Clock-drawing test, 6-CIT (6-item Cognitive Impairment Test), TYM (Test Your Memory), MoCa (Montreal Cognitive Assessment), ACE-R (Addenbrookes Cognitive Assessment), MIS (Memory Impairment Screen), BADLS (Bristol Activities of Daily Living Scale), Barthel Index, Functional Independence Measure, Instrumental Activities of Daily Living, IQCODE (Informant Questionnaire on Cognitive Decline in the Elderly), Neuropsychiatric Inventory, The Cohen-Mansfield Agitation Inventory, BEHAVE-AD, EuroQol, Short Form-36 and/or MBR Caregiver Strain Instrument, or any of the other tests as described in Sheehan B (Ther Adv Neurol Disord. 5(6):349-358 (2012)), the contents of which are herein incorporated by reference in their entirety.

In some embodiments, the present composition is administered as a solo therapeutic or as combination therapeutic for the treatment of a neurological disease/disorder or a neurodegenerative disorder, a muscular disorder or neuromuscular disorder, and/or a neuro-oncological disorder.

The AAV particles (e.g., an AAV particle comprising an AAV capsid variant) encoding the target protein may be used in combination with one or more other therapeutic agents. In some embodiments, compositions can be administered concurrently with, prior to, or subsequent to, additional therapeutic or medical procedures. In general, each agent will be administered at a dose and/or on a time schedule determined for that agent.

Therapeutic agents that may be used in combination with the AAV particles of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) can be small molecule compounds which are antioxidants, anti-inflammatory agents, anti-apoptosis agents, calcium regulators, anti-glutamatergic agents, structural protein inhibitors, compounds involved in muscle function, and compounds involved in metal ion regulation. As a non-limiting example, the combination therapy may be in combination with one or more neuroprotective agents such as small molecule compounds, growth factors and hormones which have been tested for their neuroprotective effect on motor neuron degeneration.

Compounds tested for treating neurological disease which may be used in combination with the AAV particles described herein include, but are not limited to, cholinesterase inhibitors (donepezil, rivastigmine, galantamine), NMDA receptor antagonists such as memantine, anti-psychotics, anti-depressants, anti-convulsants (e.g., sodium valproate and levetiracetam for myoclonus), secretase inhibitors, amyloid aggregation inhibitors, copper or zinc modulators, BACE inhibitors, inhibitors of tau aggregation, such as Methylene blue, phenothiazines, anthraquinones, n-phenylamines or rhodamines, microtubule stabilizers such as NAP, taxol or paclitaxel, kinase or phosphatase inhibitors such as those targeting GSK3β (lithium) or PP2A, immunization with Aβ peptides or tau phospho-epitopes, anti-tau or anti-amyloid antibodies, dopamine-depleting agents (e.g., tetrabenazine for chorea), benzodiazepines (e.g., clonazepam for myoclonus, chorea, dystonia, rigidity, and/or spasticity), amino acid precursors of dopamine (e.g., levodopa for rigidity), skeletal muscle relaxants (e.g., baclofen, tizanidine for rigidity and/or spasticity), inhibitors for acetylcholine release at the neuromuscular junction to cause muscle paralysis (e.g., botulinum toxin for bruxism and/or dystonia), atypical neuroleptics (e.g., olanzapine and quetiapine for psychosis and/or irritability, risperidone, sulpiride and haloperidol for psychosis, chorea and/or irritability, clozapine for treatment-resistant psychosis, aripiprazole for psychosis with prominent negative symptoms), selective serotonin reuptake inhibitors (SSRIs) (e.g., citalopram, fluoxetine, paroxetine, sertraline, mirtazapine, venlafaxine for depression, anxiety, obsessive compulsive behavior and/or irritability), hypnotics (e.g., xopiclone and/or zolpidem for altered sleep-wake cycle), anticonvulsants (e.g., sodium valproate and carbamazepine for mania or hypomania) and mood stabilizers (e.g., lithium for mania or hypomania).

Neurotrophic factors may be used in combination therapy with the AAV particles of the present disclosure (e.g., an AAV particle comprising an AAV capsid variant) for treating neurological disease. Generally, a neurotrophic factor is defined as a substance that promotes survival, growth, differentiation, proliferation and/or maturation of a neuron, or stimulates increased activity of a neuron. In some embodiments, the present methods further comprise delivery of one or more trophic factors into the subject in need of treatment. Trophic factors may include, but are not limited to, IGF-I, GDNF, BDNF, CTNF, VEGF, Colivelin, Xaliproden, Thyrotrophin-releasing hormone and ADNF, and variants thereof.

In one aspect, the AAV particle described herein (e.g., an AAV particle comprising an AAV capsid variant) may be co-administered with AAV particles expressing neurotrophic factors such as AAV-IGF-I (See e.g., Vincent et al., Neuromolecular medicine, 2004, 6, 79-85; the contents of which are incorporated herein by reference in their entirety) and AAV-GDNF (See e.g., Wang et al., J Neurosci., 2002, 22, 6920-6928; the contents of which are incorporated herein by reference in their entirety).

In some embodiments, administration of the AAV particles (e.g., an AAV particle comprising an AAV capsid variant) to a subject will modulate, e.g., increase or decrease, the expression of a target protein in a subject and the modulation, e.g., increase or decrease of the presence, level, activity, and/or expression of the target protein will reduce the effects and/or symptoms of a neurological disease/disorder or a neurodegenerative disorder, a muscular disorder or neuromuscular disorder, and/or a neuro-oncological disorder in a subject.

Definitions

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains.

Articles such as “a,” “an,” and “the” may mean one or more than one unless indicated to the contrary or otherwise evident from the context. Claims or descriptions that include “or” between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process unless indicated to the contrary or otherwise evident from the context. The disclosure includes embodiments in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process. The disclosure includes embodiments in which more than one, or the entire group members are present in, employed in, or otherwise relevant to a given product or process.

It is also noted that the term “comprising” is intended to be open and permits but does not require the inclusion of additional elements or steps. When the term “comprising” is used herein, the term “consisting of” and “consisting essentially thereof” is thus also encompassed and disclosed.

Where ranges are given, endpoints are included. Furthermore, it is to be understood that unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or subrange within the stated ranges in different embodiments of the disclosure, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

Adeno-associated virus: As used herein, the term “adeno-associated virus” or “AAV” refers to members of the dependovirus genus or a variant, e.g., a functional variant, thereof. In some embodiments, the AAV is wildtype, or naturally occurring. In some embodiments, the AAV is recombinant.

AAV Particle: As used herein, an “AAV particle” refers to a particle or a virion comprising an AAV capsid, e.g., an AAV capsid variant, and a polynucleotide, e.g., a viral genome or a vector genome. In some embodiments, the viral genome of the AAV particle comprises at least one payload region and at least one ITR. In some embodiments, an AAV particle of the disclosure is an AAV particle comprising an AAV variant. In some embodiments, the AAV particle is capable of delivering a nucleic acid, e.g., a payload region, encoding a payload to cells, typically, mammalian, e.g., human, cells. In some embodiments, an AAV particle of the present disclosure may be produced recombinantly. In some embodiments, an AAV particle may be derived from any serotype, described herein or known in the art, including combinations of serotypes (e.g., “pseudotyped” AAV) or from various genomes (e.g., single stranded or self-complementary). In some embodiments, the AAV particle may be replication defective and/or targeted. It is to be understood that reference to the AAV particle of the disclosure also includes pharmaceutical compositions thereof, even if not explicitly recited.

Administering: As used herein, the term “administering” refers to providing a pharmaceutical agent or composition to a subject.

Amelioration: As used herein, the term “amelioration” or “ameliorating” refers to a lessening of severity of at least one indicator of a condition or disease. For example, in the context of neurodegeneration disorder, amelioration includes the reduction of neuron loss.

Amplicon: As used herein, “amplicon” may refer to any piece of RNA or DNA formed as the product of amplification events, e.g. PCR. In some embodiments, full-length capsid amplicons may be used as templates for next generation sequencing (NGS) library generation. Full-length capsid amplicons may be used for cloning into a DNA library for any number of additional rounds of AAV selection as described herein.

Animal: As used herein, the term “animal” refers to any member of the animal kingdom. In some embodiments, “animal” refers to humans at any stage of development. In some embodiments, “animal” refers to non-human animals at any stage of development. In certain embodiments, the non-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, cattle, a primate, or a pig). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, and worms. In some embodiments, the animal is a transgenic animal, genetically engineered animal, or a clone.

Antisense strand: As used herein, the term “the antisense strand” or “the first strand” or “the guide strand” of a siRNA molecule refers to a strand that is substantially complementary to a section of about 10-50 nucleotides, e.g., about 15-30, 16-25, 18-23 or 19-22 nucleotides of the mRNA of a gene targeted for silencing. The antisense strand or first strand has sequence sufficiently complementary to the desired target mRNA sequence to direct target-specific silencing, e.g., complementarity sufficient to trigger the destruction of the desired target mRNA by the RNAi machinery or process.

Approximately: As used herein, the term “approximately” or “about,” as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In certain embodiments, the term “approximately” or “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

Biopanning: As used herein, the term “biopanning” refers to an AAV capsid library selection process comprising administration of an AAV particle with enhanced tissue- and/or cell type-specific transduction to a cell and/or subject; extraction of nucleotides encoded by said AAV particle from said transduced tissue- and/or cell type-specific; and, use of the extracted nucleotides for cloning into a nucleotide library for the generation of AAV particles for subsequent rounds of the same.

Capsid: As used herein, the term “capsid” refers to the exterior, e.g., a protein shell, of a virus particle, e.g., an AAV particle, that is substantially (e.g., >50%, >60%, >70%, >80%, >90%, >95%, >99%, or 100%) protein. In some embodiments, the capsid is an AAV capsid comprising an AAV capsid protein described herein, e.g., a VP1, VP2, and/or VP3 polypeptide. The AAV capsid protein can be a wild-type AAV capsid protein or a variant, e.g., a structural and/or functional variant from a wild-type or a reference capsid protein, referred to herein as an “AAV capsid variant.” In some embodiments, the AAV capsid variant described herein has the ability to enclose, e.g., encapsulate, a viral genome and/or is capable of entry into a cell, e.g., a mammalian cell. In some embodiments, the AAV capsid variant described herein may have modified tropism compared to that of a wild-type AAV capsid, e.g., the corresponding wild-type capsid.

Complementary and substantially complementary: As used herein, the term “complementary” refers to the ability of polynucleotides to form base pairs with one another. Base pairs are typically formed by hydrogen bonds between nucleotide units in antiparallel polynucleotide strands. Complementary polynucleotide strands can form base pairs in the Watson-Crick manner (e.g., A to T, A to U, C to G), or in any other manner that allows for the formation of duplexes. As persons skilled in the art are aware, when using RNA as opposed to DNA, uracil rather than thymine is the base that is considered to be complementary to adenine. However, when a U is denoted in the context of the present disclosure, the ability to substitute a T is implied, unless otherwise stated. Perfect complementarity or 100% complementarity refers to the situation in which each nucleotide unit of one polynucleotide strand can form a hydrogen bond with a nucleotide unit of a second polynucleotide strand. Less than perfect complementarity refers to the situation in which some, but not all, nucleotide units of two strands can form hydrogen bond with each other. For example, for two 20-mers, if only two base pairs on each strand can form a hydrogen bond with each other, the polynucleotide strands exhibit 10% complementarity. In the same example, if 18 base pairs on each strand can form hydrogen bonds with each other, the polynucleotide strands exhibit 90% complementarity. The term “complementary” as used herein can encompass fully complementary, partially complementary, or substantially complementary. As used herein, the term “substantially complementary” means that the siRNA has a sequence (e.g., in the antisense strand) which is sufficient to bind the desired target mRNA, and to trigger the RNA silencing of the target mRNA. “Fully complementary”, “perfect complementarity”, or “100% complementarity” refers to the situation in which each nucleotide unit of one polynucleotide or oligonucleotide strand can base-pair with a nucleotide unit of a second polynucleotide or oligonucleotide strand.

Control Elements: As used herein, “control elements”, “regulatory control elements” or “regulatory sequences” refers to promoter regions, polyadenylation signals, transcription termination sequences, upstream regulatory domains, origins of replication, internal ribosome entry sites (“IRES”), enhancers, and the like, which provide for the replication, transcription and translation of a coding sequence in a recipient cell. Not all of these control elements need always be present as long as the selected coding sequence is capable of being replicated, transcribed and/or translated in an appropriate host cell.

Delivery: As used herein, “delivery” refers to the act or manner of delivering an AAV particle, a compound, substance, entity, moiety, cargo or payload.

Element: As used herein, the term “element” refers to a distinct portion of an entity. In some embodiments, an element may be a polynucleotide sequence with a specific purpose, incorporated into a longer polynucleotide sequence.

Encapsulate: As used herein, the term “encapsulate” means to enclose, surround or encase. As an example, a capsid protein, e.g., an AAV capsid variant, often encapsulates a viral genome. In some embodiments, encapsulate within a capsid, e.g., an AAV capsid variant, encompasses 100% coverage by a capsid, as well as less than 100% coverage, e.g., 95%, 90%, 85%, 80%, 70%, 60% or less. For example, gaps or discontinuities may be present in the capsid so long as the viral genome is retained in the capsid, e.g., prior to entry into a cell.

Effective Amount: As used herein, the term “effective amount” of an agent is that amount sufficient to effect beneficial or desired results, for example, clinical results, and, as such, an “effective amount” depends upon the context in which it is being applied. For example, in the context of administering an agent that treats cancer, an effective amount of an agent is, for example, an amount sufficient to achieve treatment, as defined herein, of cancer, as compared to the response obtained without administration of the agent.

Expression: As used herein, “expression” of a nucleic acid sequence refers to one or more of the following events: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of an RNA transcript (e.g., by splicing, editing, 5′ cap formation, and/or 3′ end processing); (3) translation of an RNA into a polypeptide or protein; and (4) post-translational modification of a polypeptide or protein.

Formulation: As used herein, a “formulation” includes at least one AAV particle (active ingredient) and an excipient, and/or an inactive ingredient.

Fragment: A “fragment,” as used herein, refers to a portion. For example, an antibody fragment may comprise a CDR, or a heavy chain variable region, or a scFv, etc.

Homology: As used herein, the term “homology” refers to the overall relatedness between polymeric molecules, e.g. between polynucleotide molecules (e.g. DNA molecules and/or RNA molecules) and/or between polypeptide molecules. In some embodiments, polymeric molecules are considered to be “homologous” to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical or similar. The term “homologous” necessarily refers to a comparison between at least two sequences (polynucleotide or polypeptide sequences). In accordance with the disclosure, two polynucleotide sequences are considered to be homologous if the polypeptides they encode are at least about 50%, 60%, 70%, 80%, 90%, 95%, or even 99% for at least one stretch of at least about 20 amino acids. In some embodiments, homologous polynucleotide sequences are characterized by the ability to encode a stretch of at least 4-5 uniquely specified amino acids. For polynucleotide sequences less than 60 nucleotides in length, homology is determined by the ability to encode a stretch of at least 4-5 uniquely specified amino acids. In accordance with the disclosure, two protein sequences are considered to be homologous if the proteins are at least about 50%, 60%, 70%, 80%, or 90% identical for at least one stretch of at least about 20 amino acids.

Identity: As used herein, the term “identity” refers to the overall relatedness between polymeric molecules, e.g., between polynucleotide molecules (e.g. DNA molecules and/or RNA molecules) and/or between polypeptide molecules. Calculation of the percent identity of two polynucleotide sequences, for example, can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second nucleic acid sequences for optimal alignment and non-identical sequences can be disregarded for comparison purposes). In certain embodiments, the length of a sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or 100% of the length of the reference sequence. The nucleotides at corresponding nucleotide positions are then compared. When a position in the first sequence is occupied by the same nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent identity between two nucleotide sequences can be determined using methods such as those described in Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; Computer Analysis of Sequence Data, Part I, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991; the contents of each of which are incorporated herein by reference in their entirety. For example, the percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller (CABIOS, 1989, 4:11-17), which has been incorporated into the ALIGN program (version 2.0) using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. The percent identity between two nucleotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna.CMP matrix. Methods commonly employed to determine percent identity between sequences include, but are not limited to those disclosed in Carillo, H., and Lipman, D., SIAM J Applied Math., 48:1073 (1988); incorporated herein by reference. Techniques for determining identity are codified in publicly available computer programs. Exemplary computer software to determine homology between two sequences include, but are not limited to, GCG program package, Devereux, J., et al., Nucleic Acids Research, 12(1), 387 (1984)), BLASTP, BLASTN, and FASTA Altschul, S. F. et al., J. Molec. Biol., 215, 403 (1990)).

Inhibit expression of a gene: As used herein, the phrase “inhibit expression of a gene” means to cause a reduction in the amount of an expression product of the gene. The expression product can be an RNA transcribed from the gene (e.g., an mRNA) or a polypeptide translated from an mRNA transcribed from the gene. Typically, a reduction in the level of an mRNA results in a reduction in the level of a polypeptide translated therefrom. The level of expression may be determined using standard techniques for measuring mRNA or protein.

Inverted terminal repeat: As used herein, the term “inverted terminal repeat” or “ITR” refers to a cis-regulatory element for the packaging of polynucleotide sequences into viral capsids.

Isolated: As used herein, the term “isolated” refers to a substance or entity that is altered or removed from the natural state, e.g., altered or removed from at least some of component with which it is associated in the natural state. For example, a nucleic acid or a peptide naturally present in a living animal is not “isolated,” but the same nucleic acid or peptide partially or completely separated from the coexisting materials of its natural state is “isolated.” An isolated nucleic acid or protein can exist in substantially purified form, or can exist in a non-native environment such as, for example, a host cell. Such polynucleotides could be part of a vector and/or such polynucleotides or polypeptides could be part of a composition, and still be isolated in that such vector or composition is not part of the environment in which it is found in nature. In some embodiments, an isolated nucleic acid is recombinant, e.g., incorporated into a vector.

Library: As used herein, the term “library” refers to a diverse collection of linear polypeptides, polynucleotides, viral particles, or viral vectors. As examples, a library may be a DNA library or an AAV capsid library.

Molecular scaffold: As used herein a “molecular scaffold” is a framework or starting molecule that forms the sequence or structural basis against which to design or make a subsequent molecule.

Neurological disease: As used herein, a “neurological disease” is any disease associated with the central or peripheral nervous system and components thereof (e.g., neurons).

Orthogonal evolution: As used herein, the term “orthogonal evolution” refers to a method wherein AAV particles are administered for a first round of AAV selection as described herein across a set of any number of cell- and/or subject-types that may be from different species and/or strains, and wherein any number of additional, i.e., subsequent, AAV selection rounds are performed either across a set of any number of cell- and/or subject-types that may be from different species and/or strains, or across a set of any number of cell- and/or subject-types that may be from the same species and/or strain.

Open reading frame: As used herein, “open reading frame” or “ORF” refers to a sequence which does not contain a stop codon in a given reading frame.

Particle: As used herein, a “particle” is a virus comprised of at least two components, a protein capsid and a polynucleotide sequence enclosed within the capsid.

Payload region: As used herein, a “payload region” is any nucleic acid sequence (e.g., within the viral genome) which encodes one or more “payloads” of the disclosure. As non-limiting examples, a payload region may be a nucleic acid sequence within the viral genome of an AAV particle, which encodes a payload, wherein the payload is an RNAi agent or a polypeptide. Payloads of the present disclosure may be, but are not limited to, peptides, polypeptides, proteins, antibodies, RNAi agents, etc.

Polypeptide: As used herein, “polypeptide” means a polymer of amino acid residues (natural or unnatural) linked together most often by peptide bonds. The term, as used herein, refers to proteins, polypeptides, and peptides of any size, structure, or function. In some instances, the polypeptide encoded is smaller than about 50 amino acids and the polypeptide is then termed a peptide. If the polypeptide is a peptide, it will be at least about 2, 3, 4, or at least 5 amino acid residues long. Thus, polypeptides include gene products, naturally occurring polypeptides, synthetic polypeptides, homologs, orthologs, paralogs, fragments and other equivalents, variants, and analogs of the foregoing. A polypeptide may be a single molecule or may be a multi-molecular complex such as a dimer, trimer or tetramer. They may also comprise single chain or multichain polypeptides and may be associated or linked. The term polypeptide may also apply to amino acid polymers in which one or more amino acid residues are an artificial chemical analogue of a corresponding naturally occurring amino acid.

Polypeptide variant: The term “polypeptide variant” refers to molecules which differ in their amino acid sequence from a native or reference sequence. The amino acid sequence variants may possess substitutions, deletions, and/or insertions at certain positions within the amino acid sequence, as compared to a native or reference sequence. In some embodiments, a variant comprises a sequence having at least about 50%, at least about 80%, or at least about 90%, identical (homologous) to a native or a reference sequence.

Peptide: As used herein, “peptide” is less than or equal to 50 amino acids long, e.g., about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 amino acids long.

Pharmaceutically acceptable: The phrase “pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

Preventing: As used herein, the term “preventing” or “prevention” refers to partially or completely delaying onset of an infection, disease, disorder and/or condition; partially or completely delaying onset of one or more symptoms, features, or clinical manifestations of a particular infection, disease, disorder, and/or condition; partially or completely delaying onset of one or more symptoms, features, or manifestations of a particular infection, disease, disorder, and/or condition; partially or completely delaying progression from an infection, a particular disease, disorder and/or condition; and/or decreasing the risk of developing pathology associated with the infection, the disease, disorder, and/or condition.

Prophylactic: As used herein, “prophylactic” refers to a therapeutic or course of action used to prevent the spread of disease.

Prophylaxis: As used herein, a “prophylaxis” refers to a measure taken to maintain health and prevent the spread of disease.

Region: As used herein, the term “region” refers to a zone or general area. In some embodiments, when referring to a protein or protein module, a region may comprise a linear sequence of amino acids along the protein or protein module or may comprise a three-dimensional area, an epitope and/or a cluster of epitopes. In some embodiments, regions comprise terminal regions. As used herein, the term “terminal region” refers to regions located at the ends or termini of a given agent. When referring to proteins, terminal regions may comprise N- and/or C-termini.

In some embodiments, when referring to a polynucleotide, a region may comprise a linear sequence of nucleic acids along the polynucleotide or may comprise a three-dimensional area, secondary structure, or tertiary structure. In some embodiments, regions comprise terminal regions. As used herein, the term “terminal region” refers to regions located at the ends or termini of a given agent. When referring to polynucleotides, terminal regions may comprise 5′ and/or 3′ termini.

RNA or RNA molecule: As used herein, the term “RNA” or “RNA molecule” or “ribonucleic acid molecule” refers to a polymer of ribonucleotides; the term “DNA” or “DNA molecule” or “deoxyribonucleic acid molecule” refers to a polymer of deoxyribonucleotides. DNA and RNA can be synthesized naturally, e.g., by DNA replication and transcription of DNA, respectively; or be chemically synthesized. DNA and RNA can be single-stranded (i.e., ssRNA or ssDNA, respectively) or multi-stranded (e.g., double stranded, i.e., dsRNA and dsDNA, respectively). The term “mRNA” or “messenger RNA”, as used herein, refers to a single stranded RNA that encodes the amino acid sequence of one or more polypeptide chains.

RNA interfering or RNAi: As used herein, the term “RNA interfering” or “RNAi” refers to a sequence specific regulatory mechanism mediated by RNA molecules which results in the inhibition or interfering or “silencing” of the expression of a corresponding protein-coding gene. RNAi has been observed in many types of organisms, including plants, animals and fungi. RNAi occurs in cells naturally to remove foreign RNAs (e.g., viral RNAs). Natural RNAi proceeds via fragments cleaved from free dsRNA which direct the degradative mechanism to other similar RNA sequences. RNAi is controlled by the RNA-induced silencing complex (RISC) and is initiated by short/small dsRNA molecules in cell cytoplasm, where they interact with the catalytic RISC component argonaute. The dsRNA molecules can be introduced into cells exogenously. Exogenous dsRNA initiates RNAi by activating the ribonuclease protein Dicer, which binds and cleaves dsRNAs to produce double-stranded fragments of 21-25 base pairs with a few unpaired overhang bases on each end. These short double stranded fragments are called small interfering RNAs (siRNAs).

RNAi agent: As used herein, the term “RNAi agent” refers to an RNA molecule, or its derivative, that can induce inhibition, interfering, or “silencing” of the expression of a target gene and/or its protein product. An RNAi agent may knock-out (virtually eliminate or eliminate) expression, or knock-down (lessen or decrease) expression. The RNAi agent may be, but is not limited to, dsRNA, siRNA, shRNA, pre-miRNA, pri-miRNA, miRNA, stRNA, lncRNA, piRNA, or snoRNA.

miR binding site: As used herein, a “miR binding site” comprises a nucleic acid sequence (whether RNA or DNA, e.g., differ by “U” of RNA or “T” in DNA) that is capable of binding, or binds, in whole or in part to a microRNA (miR) through complete or partial hybridization. Typically, such binding occurs between the miR and the miR binding site in the reverse complement orientation. In some embodiments, the miR binding site is transcribed from the AAV vector genome encoding the miR binding site.

In some embodiments, a miR binding site may be encoded or transcribed in series. Such a “miR binding site series” or “miR BSs” may include two or more miR binding sites having the same or different nucleic acid sequence.

Spacer: As used here, a “spacer” is generally any selected nucleic acid sequence of, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides in length, which is located between two or more consecutive miR binding site sequences. Spacers may also be more than 10 nucleotides in length, e.g., 20, 30, 40, or 50 or more than 50 nucleotides.

Sample: As used herein, the term “sample” or “biological sample” refers to a subset of its tissues, cells, nucleic acids, or component parts (e.g. body fluids, including but not limited to blood, serum, mucus, lymphatic fluid, synovial fluid, cerebrospinal fluid, saliva, amniotic fluid, amniotic cord blood, urine, vaginal fluid and semen).

Self-complementary viral particle: As used herein, a “self-complementary viral particle” is a particle comprised of at least two components, a protein capsid and a self-complementary viral genome enclosed within the capsid.

Sense Strand: As used herein, the term “the sense strand” or “the second strand” or “the passenger strand” of a siRNA molecule refers to a strand that is complementary to the antisense strand or first strand. The antisense and sense strands of a siRNA molecule are hybridized to form a duplex structure. As used herein, a “siRNA duplex” includes a siRNA strand having sufficient complementarity to a section of about 10-50 nucleotides of the mRNA of the gene targeted for silencing and a siRNA strand having sufficient complementarity to form a duplex with the other siRNA strand.

Similarity: As used herein, the term “similarity” refers to the overall relatedness between polymeric molecules, e.g. between polynucleotide molecules (e.g. DNA molecules and/or RNA molecules) and/or between polypeptide molecules. Calculation of percent similarity of polymeric molecules to one another can be performed in the same manner as a calculation of percent identity, except that calculation of percent similarity takes into account conservative substitutions as is understood in the art.

Short interfering RNA or siRNA: As used herein, the terms “short interfering RNA,” “small interfering RNA” or “siRNA” refer to an RNA molecule (or RNA analog) comprising between about 5-60 nucleotides (or nucleotide analogs) which is capable of directing or mediating RNAi. Preferably, a siRNA molecule comprises between about 15-30 nucleotides or nucleotide analogs, such as between about 16-25 nucleotides (or nucleotide analogs), between about 18-23 nucleotides (or nucleotide analogs), between about 19-22 nucleotides (or nucleotide analogs) (e.g., 19, 20, 21 or 22 nucleotides or nucleotide analogs), between about 19-25 nucleotides (or nucleotide analogs), and between about 19-24 nucleotides (or nucleotide analogs). The term “short” siRNA refers to a siRNA comprising 5-23 nucleotides, preferably 21 nucleotides (or nucleotide analogs), for example, 19, 20, 21 or 22 nucleotides. The term “long” siRNA refers to a siRNA comprising 24-60 nucleotides, preferably about 24-25 nucleotides, for example, 23, 24, 25 or 26 nucleotides. Short siRNAs may, in some instances, include fewer than 19 nucleotides, e.g., 16, 17 or 18 nucleotides, or as few as 5 nucleotides, provided that the shorter siRNA retains the ability to mediate RNAi. Likewise, long siRNAs may, in some instances, include more than 26 nucleotides, e.g., 27, 28, 29, 30, 35, 40, 45, 50, 55, or even 60 nucleotides, provided that the longer siRNA retains the ability to mediate RNAi or translational repression absent further processing, e.g., enzymatic processing, to a short siRNA. siRNAs can be single stranded RNA molecules (ss-siRNAs) or double stranded RNA molecules (ds-siRNAs) comprising a sense strand and an antisense strand which hybridized to form a duplex structure called an siRNA duplex.

Subject: As used herein, the term “subject” or “patient” refers to any organism to which a composition in accordance with the disclosure may be administered, e.g., for experimental, diagnostic, prophylactic, and/or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans) and/or plants.

Substantially: As used herein, the term “substantially” refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest. One of ordinary skill in the biological arts will understand that biological and chemical phenomena rarely, if ever, go to completion and/or proceed to completeness or achieve or avoid an absolute result. The term “substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

Target Cells: As used herein, “target cells” or “target tissue” refers to any one or more cells of interest. The cells may be found in vitro, in vivo, in situ or in the tissue or organ of an organism. The organism may be an animal, preferably a mammal, more preferably a human and most preferably a patient.

Therapeutic Agent: The term “therapeutic agent” refers to any agent that, when administered to a subject, has a therapeutic, diagnostic, and/or prophylactic effect and/or elicits a desired biological and/or pharmacological effect.

Therapeutically effective amount: As used herein, the term “therapeutically effective amount” means an amount of an agent to be delivered (e.g., nucleic acid, drug, therapeutic agent, diagnostic agent, prophylactic agent, etc.) that is sufficient, when administered to a subject suffering from or susceptible to an infection, disease, disorder, and/or condition, to treat, improve symptoms of, diagnose, prevent, and/or delay the onset of the infection, disease, disorder, and/or condition. In some embodiments, a therapeutically effective amount is provided in a single dose.

Therapeutically effective outcome: As used herein, the term “therapeutically effective outcome” means an outcome that is sufficient in a subject suffering from or susceptible to an infection, disease, disorder, and/or condition, to treat, improve symptoms of, diagnose, prevent, and/or delay the onset of the infection, disease, disorder, and/or condition.

Treating: As used herein, the term “treating” refers to partially or completely alleviating, ameliorating, improving, relieving, delaying onset of, inhibiting progression of, reducing severity of, and/or reducing incidence of one or more symptoms or features of a particular infection, disease, disorder, and/or condition. For example, “treating” cancer may refer to inhibiting survival, growth, and/or spread of a tumor. Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and/or condition and/or to a subject who exhibits only early signs of a disease, disorder, and/or condition for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and/or condition.

Conservative amino acid substitution: As used herein, a “conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

Variant: As used herein, the term “variant” refers to a polypeptide or polynucleotide that has an amino acid or a nucleotide sequence that is substantially identical, e.g., having at least 70%, 75%, 80%, 85%, 90%, 95% or 99% sequence identity to a reference sequence. In some embodiments, the variant is a functional variant.

Functional Variant: As used herein, the term “functional variant” refers to a polypeptide variant or a polynucleotide variant that has at least one activity of the reference sequence.

Insertional Variant: “Insertional variants” when referring to polypeptides are those with one or more amino acids inserted, e.g., immediately adjacent or subsequent, to a position in an amino acid sequence. “Immediately adjacent” or “immediately subsequent” to an amino acid means connected to either the alpha-carboxy or alpha-amino functional group of the amino acid.

Deletional Variant: “Deletional variants” when referring to polypeptides, are those with one or more amino acids in deleted from a reference protein.

Vector: As used herein, the term “vector” refers to any molecule or moiety which transports, transduces or otherwise acts as a carrier of a heterologous molecule. In some embodiments, vectors may be plasmids. In some embodiments, vectors may be viruses. An AAV particle is an example of a vector. Vectors of the present disclosure may be produced recombinantly and may be based on and/or may comprise adeno-associated virus (AAV) parent or reference sequences. The heterologous molecule may be a polynucleotide and/or a polypeptide.

Viral Genome: As used herein, the terms “viral genome” or “vector genome” refer to the nucleic acid sequence(s) encapsulated in an AAV particle. A viral genome comprises a nucleic acid sequence with at least one payload region encoding a payload and at least one ITR.

Equivalents and Scope

The disclosures of each and every patent, patent application, and publication cited herein are hereby incorporated herein by reference in their entirety. While this invention has been disclosed with reference to certain embodiments, it is apparent that further embodiments and variations of this invention may be devised by others skilled in the art without departing from the true spirit and scope of the invention. The appended claims are intended to be construed to include all such embodiments and equivalent variations.

The present disclosure is further illustrated by the following non-limiting examples.

EXAMPLES
Example 1. High-throughput Screen of TRACER AAV Library in NHP and Mice

A TRACER based method as described in WO2020072683, WO 2021/202651, and WO2021230987, the contents of which are herein incorporated by reference in their entirety, was used to generate the AAV capsid variants described herein. An orthogonal evolution approach was combined with a high throughput screening by NGS. Briefly, the library of AAV capsid variants was generated using a sliding window approach, where 6 amino acid sequences were inserted into 8 different positions across loop IV of AAV9, including immediately subsequent to positions 453, 454, 455, 456, 457, 458, 459, and 460, relative to a reference sequence numbered according to SEQ ID NO: 138. The initial library was passed twice through non-human primates (NHP, 2-4 years of age). After the second passage (e.g., 28 days post injection into two NHPs), RNA was extracted from six brain regions. Following RNA recovery and RT-PCR amplification, a systematic NGS enrichment analysis was performed to calculate fold enrichment relative to an AAV9 wild-type control. Following these two passages, approximately 21195 variants were identified with an average fold change greater than wild-type. Of the 21195 variants, 1558 demonstrated a fold-change of greater than 6 compared to wild-type and were detected across all brain regions investigated. Of these 1558, approximately 1470 variants were selected for constructing a synthetic library and a third passage through two NHPs. Within the 1470 variants selected for further characterization and investigation, there was a relatively even distribution for each insertion position of the sliding window used to generate the initial library.

After creation of the synthetic library with the sub-selected variants, the synthetic library was screened (passage 3) in two NHPs (2-4 years of age) and two strains of mice, BALB/c (n=3, 6-8 weeks of age) and C57B1/6 mice (n=3, 6-8 weeks of age), in a first cross-species evolution screen. The animals were injected intravenously with the synthetic library. After a period in vivo, (e.g., 28-days) RNA was extracted from nervous tissue, e.g., brain, spinal cord, and DRG of the NHPs and the brains of mice. Following RNA recovery and RT-PCR amplification, a systematic NGS enrichment analysis was performed, and the peptides comprised within the variants were identified and the capsid enrichment ratio for each variant compared to the wild-type AAV9 control was calculated (fold enrichment relative to wild-type AAV9) (Table 9). Values above 1 indicate an increase in expression relative to AAV9. All animals were dosed intravenously at 2-3 VG/kg across the screen.

As shown in Table 9, approximately 700 variants demonstrated an increase in expression relative to AAV9, and several variants demonstrated a greater than 10-fold enrichment relative to AAV9 in the brain of NHPs. Further, the variants demonstrating the greatest fold enrichment in the brain also demonstrated the greatest fold enrichment in the spinal cord relative to AAV9 in NHPs. These variants also demonstrated de-targeting in the DRG (data not shown). For instance, the variant comprising GSGSPHSKAQNQQT (SEQ ID NO: 200) demonstrated a 76.6 fold enrichment in the brain, a 29.4 fold enrichment in the spinal cord, and 0.4 fold enrichment in the DRG of NHPs relative to AAV9; and GHDSPHKSGQNQQT (SEQ ID NO: 201) demonstrated a 62.6 fold enrichment in the brain, a 15.6 fold enrichment in the spinal cord, and 0.0 fold enrichment in the DRG of NHPs relative to AAV9. Also, across the peptides comprised within the AAV capsid variants with the greatest fold-enrichment in the NHP brain relative wild-type AAV9, it was observed that each of these peptides comprised an SPH motif in the same position (e.g., immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138), regardless of the insertion position within the variant capsid, as well as a positive amino acid (e.g., K or R) in one of the next three residues subsequent to the SPH motif.

Those variants with the greatest fold enrichment in the brains of NHPs also had the greatest fold enrichment in the brains of both mouse species. Also, when comparing the fold enrichment relative to wild-type for each variant between the two species of mice investigated (C57B1/6 and BALB/c mice), they were highly correlated (R²=0.8591).

TABLE 9

NGS fold-enrichment of AAV capsid variants in NHPs and mice

Fold
Fold

Fold
enrichment
enrichment
Fold enrichment

SEQ
enrichment over
over AAV9 in
over AAV9 in
over AAV9 in

ID
AAV9 in the
the spinal
the brain of
the brain of Mice

Peptide Sequence
NO:
brain of NHPs
cord of NHPs
Mice (C57Bl/6)
(BALB/c)

GSGSPHSKAQNQQT
200
73.615
29.402
25.293
41.304

GHDSPHKSGQNQQT
201
62.612
15.641
63.993
49.760

GSGSPHARMQNQQT
202
56.138
22.690
7.795
4.164

GSGSPHVKSQNQQT
203
37.551
13.649
8.069
15.861

GQDSPHKSGQNQQT
204
24.569
3.548
57.344
42.615

GSGSPHASRQNQQT
205
18.265
7.804
28.028
36.577

GSGSPHASRQNKQT
206
17.520
35.029
13.096
18.114

GSGSPHVKIQNQQT
207
16.854
9.068
2.173
2.227

GSGSPHSKAKNQQT
208
14.458
0.049
21.494
23.556

GSGSPHKKNQNQQT
209
12.991
0.379
25.958
7.415

GSGSPHVRMQNQQT
210
11.574
6.764
9.121
10.076

GSGSPHASRQKQQT
211
11.417
0.005
7.413
12.400

GHSSPHRSGQNQQT
212
10.357
1.887
23.197
25.442

GMRTYHLSGQNQQT
213
9.241
1.939
2.033
1.586

GSGSPHTRGQNQQT
214
7.092
3.815
10.801
6.240

GSGIIPVSSQNQQT
215
6.352
0.000
0.642
0.253

GSEYGHKSGQNQQT
216
6.308
2.750
5.198
5.332

GRGQNVSSVHRQQT
217
5.404
0.000
1.206
0.691

GSSHRFYGGQNQQT
218
4.732
0.000
0.787
0.110

GYFVAAWSGQNQQT
219
4.488
0.000
0.071
0.175

GSVLHSHAAQNQQT
220
4.150
6.448
0.675
0.423

GSGDLVVSTQNQQT
221
3.874
1.177
0.411
0.273

GSYGMAASGQNQQT
222
3.817
10.052
1.274
0.829

GLNHFGASGQNQQT
223
3.802
3.188
0.774
0.579

GSTGSHSAGQNQHT
224
3.717
0.285
1.190
0.850

GLAGHTVSGQNQQT
225
3.632
0.229
0.972
0.202

GIILGASSGQNQQT
226
3.630
4.868
1.378
0.865

GSGVSTYNIQNQQT
227
3.609
2.912
0.769
0.520

GSLVSVQTGQNQQT
228
3.534
6.043
0.903
0.469

GQSSPHRSGQNQQT
229
3.496
2.142
12.352
19.366

GREYGHKSGQNQQT
230
3.453
0.000
1.422
0.959

GHTLTLSSGQNQQT
231
3.405
5.648
0.648
0.606

GSITLIPSGQNQQT
232
3.361
3.917
0.326
0.435

GSNGFTALGQNQQT
233
3.361
2.663
0.830
0.332

GSGHSSHSVQNQQT
234
3.339
3.318
0.942
0.424

GSGIPQRSGKNQQT
235
3.331
0.000
1.418
1.685

GSGDTLHMLQNQQT
236
3.317
1.174
0.393
0.482

GERHTVLSGQNQQT
237
3.289
3.008
1.027
0.607

GSGMPQSHIQNQQT
238
3.289
11.609
0.514
0.334

GSGQLSGIGGNQQT
239
3.266
0.287
0.993
0.626

GSGQNRKPASFAQT
240
3.204
0.000
0.892
1.061

GSGSVSQLGQNQQT
241
3.184
2.307
0.596
0.375

GSDFLGTHGQNQQT
242
3.171
0.348
1.038
0.750

GQIVQNPSGQNQQT
243
3.133
0.406
1.446
0.635

GSGTQIPSQQNQQT
244
3.112
1.224
0.470
0.151

GSGQNQQSAREGLT
245
3.111
5.632
1.221
1.104

GSGLGMSTGQNQQT
246
3.110
5.499
0.458
0.660

GSGLPVLSGQNQQT
247
3.100
4.149
0.631
0.210

GSGHSIRTDQNQQT
248
3.074
15.600
0.229
0.148

GSGQSVQTVVNQQT
249
3.057
5.441
0.582
0.240

GSGQNRAQSRFQQT
250
3.043
0.000
0.619
1.788

GGGDLGRSSQNQQT
251
3.036
4.830
0.916
0.539

GGGTKMDSGQNQQT
252
3.034
0.000
0.733
0.297

GSGSPHPSRQNQQT
253
3.017
1.993
1.869
0.975

GSGQFTNAGMNQQT
254
2.969
0.936
0.565
0.418

GGRNGHTVGQNQQT
255
2.965
3.732
1.105
1.003

GSGFGPQTGQNQQT
256
2.964
2.861
1.280
0.849

GRTDSHTSGQNQQT
257
2.913
1.510
1.299
0.704

GYEVLGSSGQNQQT
258
2.891
0.000
2.459
0.319

GSVHLSVTGQNQQT
259
2.882
1.157
0.741
0.282

GFMSYKGSGQNQQT
260
2.865
0.209
1.808
0.569

GNIAGSVSGQNQQT
261
2.849
1.187
0.446
0.257

GSGSHRDVSQNQQT
262
2.843
4.022
0.626
0.550

GGLGSMSSGQNQQT
263
2.812
1.405
1.802
0.822

GSGHLPQSAQNQQT
264
2.803
7.828
0.826
0.496

GGVLVGGSGQNQQT
265
2.778
0.178
1.527
0.688

GTHPYTSSGQNQQT
266
2.775
1.684
0.758
0.471

GSGQNQQLKENRST
267
2.765
0.062
1.149
1.118

GSGQNQQTSPHNHT
268
2.761
3.132
1.524
0.845

GSGTLYPQSQNQQT
269
2.761
5.558
0.324
0.160

GSGQNQQSNWITKT
270
2.711
0.000
0.540
0.634

GSGYTSLFLQNQQT
271
2.710
0.010
0.490
1.044

GSGVMTHVLQNQQT
272
2.692
0.347
0.370
0.533

GSVSDVRAGQNQQT
273
2.661
1.647
0.267
0.747

GSGQSHMATLNQQT
274
2.657
0.724
1.173
0.504

GSGLSVHLAQNQQT
275
2.657
1.234
0.806
0.508

GSGLSHATQQNQQT
276
2.640
7.819
1.111
0.638

GSGLSVQSGQNQQT
277
2.637
2.929
1.695
1.005

GSGHMTYREKNQQT
278
2.633
5.267
1.257
0.540

GSKGVPTPGQNQQT
279
2.625
1.292
1.452
0.459

GSGLLPLSSQNQQT
280
2.612
1.130
0.501
0.293

GNGLYAVSGQNQQT
281
2.611
9.148
0.322
0.213

GFNGSPSSGQNQQT
282
2.609
12.197
2.338
0.924

GSGQIRHSDQNQQT
283
2.600
12.884
1.170
0.320

GGQVAPSSGQNQQT
284
2.581
2.427
1.433
0.709

GSFSMHTHGQNQQT
285
2.535
0.118
1.027
0.693

GSGQNQQVIQGSNT
286
2.521
8.778
0.935
0.810

GRVLHSHAGQNQQT
287
2.513
0.826
1.294
0.908

GSGQNQQTSLQDQT
288
2.505
0.500
0.315
0.968

GSGLGRAPVQNQQT
289
2.503
2.214
0.841
0.383

GNGFSSASGQNQQT
290
2.493
0.772
0.240
0.182

GSGQMASRESNQQT
291
2.492
0.300
0.341
0.288

GPGLPNHSGQNQQT
292
2.486
1.992
1.197
0.659

GNIQWQGSGQNQQT
293
2.468
6.266
1.182
0.837

GMSAHMSSGQNQQT
294
2.456
5.255
1.310
0.947

GHSFVNRSGQNQQT
295
2.447
11.148
1.305
0.756

GRAVMDHSGQNQQT
296
2.408
3.209
0.728
0.283

GALTVMQSGQNQQT
297
2.381
0.430
0.246
0.199

GSGQRSPVLPNQQT
298
2.369
6.230
0.434
0.526

GSGQNGHLSLKQQT
299
2.362
1.896
0.718
0.270

GSLPRGTSDQNQQT
300
2.362
0.000
0.453
0.495

GVAGSLVSGQNQQT
301
2.358
7.670
1.321
1.160

GRGGIPQSGQNQQT
302
2.352
8.683
1.639
1.181

GSGQYASSIPNQQT
303
2.346
3.321
1.022
0.489

GTDFGRQSSQNQQT
304
2.346
3.196
1.021
0.797

GIFMQTPSGQNQQT
305
2.344
6.198
0.938
0.252

GSGQNQQTRLVDLT
306
2.342
9.348
1.268
0.490

GTREMPLSGQNQQT
307
2.339
2.830
1.436
0.538

GSRLVHVHGQNQQT
308
2.334
1.174
1.277
0.934

GSGRLVPNGPNQQT
309
2.314
3.925
0.639
0.411

GSGYLRESPQNQQT
310
2.311
0.878
0.331
0.677

GARIQNASGQKQQT
311
2.300
2.103
1.220
1.039

GLSNPMPSGQNQQT
312
2.280
6.033
1.190
0.829

GSTVQDTRGQNQQT
313
2.270
4.979
0.576
0.473

GPFGMPSSGQNQQT
314
2.260
2.700
0.727
0.560

GSGQNHGVLSNQQT
315
2.254
1.603
1.113
0.701

GSGYSMSQAQNQQT
316
2.250
4.479
0.519
0.329

GSGMLTHTLQNQQT
317
2.246
2.272
0.496
0.199

GRGSPHASRQNQQT
318
2.241
0.000
5.050
5.856

GLSWPSTSGQNQQT
319
2.238
0.000
0.910
0.610

GNSMERTSGQNQQT
320
2.221
4.177
1.047
0.935

GSGMSPSTLQNQQT
321
2.216
3.053
0.318
0.153

GSGHGQVLSQNQQT
322
2.213
12.133
1.880
0.661

GRGQIYSTGGNQQT
323
2.210
11.629
1.329
0.743

GVVAAHNSGQNQQT
324
2.202
1.301
1.196
1.336

GDSSLRHSGQNQQT
325
2.194
0.000
0.662
0.412

GSLVSQGAGQNQQT
326
2.188
4.414
1.436
1.246

GSLLQAHSGQNQQT
327
2.182
1.008
0.575
0.748

GSGHIYVGIQNQQT
328
2.178
6.428
0.989
0.337

GHHTTVQSGQNQQT
329
2.177
6.245
0.851
0.755

GSRQSKRNELNQQT
330
2.177
0.000
1.325
0.232

GSGQNQQHVSSPRT
331
2.176
1.279
1.847
0.938

GSSKELLWGQNQQT
332
2.163
0.000
0.506
0.883

GSLSTPSSGQNQQT
333
2.159
1.279
1.094
0.669

GSIGYAGQGQNQQT
334
2.157
4.951
1.604
0.712

GSGQNQRVSNSQQT
335
2.146
0.492
1.086
0.985

GSGYASHVQQNQQT
336
2.146
3.038
1.157
0.758

GSGEYSRSGQNQQT
337
2.145
0.745
0.617
0.205

GSVSTHSSGQNQQT
338
2.145
3.446
1.198
0.918

GSGQNQHSLGNYQT
339
2.143
1.896
1.077
0.606

GSGGLDTRGQNQQT
340
2.139
6.216
0.236
0.197

GNILHATSGQNQQT
341
2.136
0.125
1.159
0.424

GSGQSYTMTQNQQT
342
2.136
6.755
0.297
0.231

GSGQNQHSAPNSQT
343
2.134
4.143
1.187
0.731

GSGQNQQTMDHNRT
344
2.130
4.944
0.642
0.440

GSNGGVGTGQNQQT
345
2.130
0.788
1.191
1.087

GAGSIIPSGQNQQT
346
2.129
7.164
0.595
0.249

GSGQTHGGQHNQQT
347
2.125
12.251
1.448
1.098

GSNLSFQSGQNQQT
348
2.122
5.853
1.087
0.719

GATLQVHSGQNQQT
349
2.122
2.219
0.623
0.545

GSGFNQRSEQNQQT
350
2.121
4.491
1.770
0.758

GSGSLRDFDQNQQT
351
2.120
6.846
0.586
0.272

GSGDSITGKQNQQT
352
2.112
1.295
0.793
0.306

GSGQDRNIVQNQQT
353
2.112
0.229
0.454
0.632

GSGLSHSHQQNQQT
354
2.109
5.852
1.256
0.592

GSGQNQQTGMSSVK
355
2.109
4.544
1.451
0.679

GSVTHGISGQNQQT
356
2.105
4.542
1.135
0.789

GVVAHQPSGQNQQT
357
2.103
0.152
0.910
2.267

GSGPILGQLQNQQT
358
2.097
2.058
0.470
0.123

GSGHVPNSGLNQQT
359
2.091
0.653
1.636
1.154

GDAGVRSSGQNQQT
360
2.068
3.918
1.033
1.193

GSGSQLMSLQNQQT
361
2.065
3.559
0.563
0.172

GSGLDYSQRQNQQT
362
2.056
0.837
0.484
0.217

GSGQSSGRLINKQT
363
2.055
28.135
0.543
0.277

GSSVSPSSGQNQQT
364
2.054
0.579
1.064
0.787

GSGQVVGLSGNQQT
365
2.052
7.212
0.785
0.881

GSNMGVPLGQNQQT
366
2.049
2.448
0.334
0.420

GSFYPSSTGQNQQT
367
2.047
2.374
0.420
0.277

GSGQNQQTRLTDLT
368
2.046
8.470
0.910
0.776

GPTNGRSSGQNQQT
369
2.034
8.903
0.936
1.308

GSGLLHGKLQNQQT
370
2.032
2.521
1.068
0.917

GANMGHVSGQNQQT
371
2.020
0.810
1.302
1.138

GSGQNQQSGRGDLT
372
2.019
6.919
0.524
1.152

GSHGHYASGQKQQT
373
2.016
0.000
0.895
0.685

GSGDLRISPQNQQT
374
2.012
16.207
0.620
0.237

GSGMPVILGQNQQT
375
2.005
0.150
0.840
0.287

GRGVITSSGQNHQT
376
2.004
0.864
1.656
0.669

GSGHSVSGPQNQQT
377
1.993
6.259
1.370
0.619

GSRNGHTVGRNQQT
378
1.993
0.000
1.162
0.367

GAGVHMVSGQNQQT
379
1.987
6.488
1.055
0.791

GSGQNHRPSVLQQT
380
1.983
5.582
0.433
0.582

GSGSPRDSIQNQQT
381
1.981
4.914
0.171
0.446

GSGQGIHSSVNQQT
382
1.981
4.873
0.632
0.634

GSGQQLSITPNQQT
383
1.979
10.280
0.845
0.201

GGYHSQTSGQNQQT
384
1.978
2.642
1.740
1.525

GSLHHDNHGQNQQT
385
1.976
0.980
0.968
0.463

GIMARDSSGQNQQT
386
1.972
3.486
1.320
0.904

GVVHITNSGQNQQT
387
1.969
0.504
0.794
0.846

GSGQNQHSAPFNQT
388
1.969
0.499
0.759
0.870

GSGQTSGLKQNQQT
389
1.968
3.927
0.394
0.334

GSGQNQQTSLSNTA
390
1.959
1.186
1.567
1.182

GSGQNQAVHNKSQT
391
1.956
3.791
1.465
1.083

GVHTHLPSGQNQQT
392
1.952
1.364
1.414
0.796

GHLTMHNSGQNHQT
393
1.938
1.798
1.030
0.586

GSGSSSRPYQNQQT
394
1.934
3.823
0.962
0.496

GILLATPSGQNQQT
395
1.931
8.205
1.341
0.288

GSGQNAGSFPNQQT
396
1.928
12.575
1.091
0.286

GSRDGHTVGQNQQT
397
1.928
7.089
0.495
0.661

GSLLISTSGQNQQT
398
1.919
5.763
1.488
0.808

GSGAMPSHGQNQQT
399
1.915
0.000
1.142
0.912

GALVSPISGQNQQT
400
1.912
1.051
0.640
0.347

GSLSSHGVGQNQQT
401
1.911
7.498
1.218
0.804

GSGQNQQASLAMRT
402
1.910
3.577
2.066
1.638

GPGLGSHSGQNQQT
403
1.906
14.563
0.880
1.195

GHDSQHKSGQNQQT
404
1.904
6.988
1.154
0.869

GSGLTLSATQNQQT
405
1.901
0.193
0.708
0.340

GSGQVVAHVGNQQT
406
1.901
0.833
0.800
0.321

GSGLRTMTTQNQQT
407
1.900
8.939
0.838
0.594

GSGQVGRLLQNQQT
408
1.899
1.762
0.773
0.748

GSGQLSHQSVNQQT
409
1.898
4.032
0.720
0.695

GSGDRYQTLQNQQT
410
1.897
1.075
0.645
0.318

GSGQNQQLKSSAQT
411
1.891
1.197
0.908
0.716

GSGQNQYSIPVAQT
412
1.891
1.194
0.511
0.297

GSGERLHLTQNQQT
413
1.885
1.456
0.387
0.245

GSGHNQQVRTAPNT
414
1.885
1.022
1.006
0.580

GGLSHVMSGQNQQT
415
1.875
0.403
0.885
0.378

GSGQSHRDVLNQQT
416
1.872
15.082
0.138
0.280

GSGQNLAGRMDQQT
417
1.864
0.085
0.362
0.295

GSGQNQQTNRGNPM
418
1.860
3.402
1.349
1.098

GSGQSYQRDHNQQT
419
1.859
8.013
0.779
0.323

GSLLSAGMGQNQHT
420
1.856
6.168
0.589
0.342

GSGQNQQTAIYRNI
421
1.854
2.207
0.818
1.437

GSGQNQQTSGTTNC
422
1.854
8.161
1.040
0.806

GMTSHSVSGQNQQT
423
1.850
2.732
0.220
0.154

GSSQSTGYQPNQQT
424
1.847
3.388
0.522
0.577

GSLKPTTLGQNQQT
425
1.840
0.476
0.175
0.220

GRMFSLGSGQNQQT
426
1.836
8.429
1.630
1.174

GSGQNQQTALGVKC
427
1.835
1.343
1.378
1.014

GAMVSHSSGQNQQT
428
1.833
8.999
0.739
0.868

GSGQNQQRNSDSVT
429
1.829
0.000
1.238
0.842

GSGQSMTLHLNQQT
430
1.827
0.991
0.721
0.248

GSGQVHQAEVNQQT
431
1.825
0.152
0.436
0.287

GSGQNQSQNHLQQT
432
1.825
0.600
1.063
0.772

GSLLTTASGQNQQT
433
1.822
0.780
0.938
0.635

GSGLIRTAAQNQQT
434
1.822
8.339
0.808
0.998

GSGQNQQTVSRQST
435
1.820
0.472
1.330
0.796

GSGQYANHGINQQT
436
1.820
5.717
0.906
0.701

GSRSTGPSGQNQQT
437
1.819
2.479
0.440
0.466

GRGVQQKLQQNQQT
438
1.817
0.000
1.974
0.823

GSGQNQQVHLSTGT
439
1.811
0.266
1.011
0.455

GSGQNQQLSAKSST
440
1.809
1.567
1.224
1.115

GSGYKAARPQNQQT
441
1.803
0.000
1.418
0.337

GSAGISPSGQNQQT
442
1.797
1.812
0.784
0.622

GSGQNRAHAFLQQT
443
1.795
0.000
1.200
1.271

GSGLSGITMQNQQT
444
1.792
14.796
0.862
0.496

GPGSAHSSGQNQQT
445
1.785
4.392
1.099
0.872

GSSHTQALGQNQQT
446
1.784
0.143
0.882
0.874

GSGVHGVSSQNQQT
447
1.781
4.519
1.504
0.951

GSSGRDMGGQNQQT
448
1.778
2.177
1.052
0.595

GERAFPTSGQNQQT
449
1.775
6.515
0.972
0.362

GGRIVSLSGQNQQT
450
1.766
4.936
1.161
0.847

GSGQNSYSHTSQQT
451
1.765
2.262
1.130
0.658

GLGYPGSSGQNQQT
452
1.763
7.090
0.929
0.577

GSGPQSHTGQNQQT
453
1.757
9.490
0.958
0.447

GSGQNQQLSRDAST
454
1.754
3.716
1.877
0.611

GSGQILHSVPNQQT
455
1.752
1.316
0.398
0.240

GSGFHTDSRQNQQT
456
1.748
4.384
7.344
0.575

GSGQSHSLATNQQT
457
1.745
2.711
1.021
0.343

GSGQNQQTLSKPWT
458
1.743
0.253
0.845
0.733

GSGHAAISQQNQQT
459
1.742
2.373
1.211
0.520

GSGQNQQQIGGNST
460
1.741
6.169
0.877
0.576

GGGPMAGSGQNQQT
461
1.735
2.815
1.049
0.372

GMRMEYQSGQNQQT
462
1.729
3.695
0.644
0.632

GSGQNQQGTLLHQT
463
1.728
2.065
1.347
1.303

GSGQNQRSSGGVQT
464
1.723
2.056
1.805
1.165

GSGQNQRGALATQT
465
1.722
1.117
0.899
0.891

GSGTVHAATQNQQT
466
1.721
1.676
0.563
0.476

GSRMTQQFGQNQQT
467
1.720
25.798
1.233
0.976

GSSSPGASGQNQQT
468
1.717
1.244
1.378
0.660

GHPSPHVSGQNQQT
469
1.713
0.416
0.551
0.488

GSGSHHASRQNQQT
470
1.712
0.451
3.073
0.584

GAVGHSYSGQNQQT
471
1.706
0.808
0.306
0.536

GSRSQYDIGQNQQT
472
1.706
0.112
0.528
0.193

GSGQGPQERGNQQT
473
1.702
1.269
0.846
0.313

GSIAHVGTGQNQQT
474
1.696
1.264
0.837
1.045

GSGQNQQKQNHGNT
475
1.695
5.349
1.538
1.340

GSGQNQQALGSQRT
476
1.695
1.934
1.419
0.562

GSGAITHMPQNQQT
477
1.695
1.681
0.647
0.411

GSGQRNPLLLNQQT
478
1.693
0.144
0.662
0.740

GSSGIPVSHQNQQT
479
1.690
3.384
0.820
0.333

GVHSLTPSGQNQQT
480
1.687
4.104
0.475
0.215

GVIVLHGSGQNQQT
481
1.682
14.166
1.074
1.098

GGTRVVDSGQNQQT
482
1.676
9.735
0.676
0.370

GSGGVTYQSQNQQT
483
1.673
7.283
0.649
0.181

GSGQNQAGHGPGQT
484
1.670
2.861
1.040
0.887

GSGQLVTSGPNQQT
485
1.669
5.271
0.964
0.433

GSGIAAQRTQNQQT
486
1.665
2.691
1.062
0.754

GSTPAGVGGQNQQT
487
1.663
2.733
0.593
0.477

GSGQNQQTSTGVHS
488
1.660
8.271
1.039
1.075

GSGQIRQLVDNQQT
489
1.657
5.529
0.314
0.272

GSLIGMQSGQNQQT
490
1.656
6.783
0.797
0.392

GSGQIKGKMDNQQT
491
1.654
2.601
1.065
1.012

GSGSDMSSWQNQQT
492
1.651
0.175
0.281
0.303

GRGQNQQHTGLATT
493
1.650
6.174
1.134
0.691

GSGQNQQTLYSSNT
494
1.642
1.044
0.664
0.368

GSGQTQVLKSNQQT
495
1.640
3.031
1.599
0.975

GSRTLSNVGQNQQT
496
1.640
3.219
0.617
0.542

GSGVQHSLPQNQQT
497
1.639
0.764
0.440
0.387

GNYLHQASGQNQQT
498
1.635
1.454
0.816
0.181

GSGGTSVHQQNQQT
499
1.629
0.000
0.585
0.195

GMDHSRPSGQNQQT
500
1.627
3.976
0.918
0.648

GSGQNQQSMGTFTT
501
1.625
0.000
1.792
0.399

GSGQNQQTPLRPPT
502
1.624
0.352
0.874
0.472

GSGQNQHHSVSQQT
503
1.623
3.700
0.605
0.334

GSGQLRSLSTNQQT
504
1.622
6.855
1.310
0.382

GSGSPRQLSQNQQT
505
1.621
0.873
0.520
0.273

GSGQNQQTTASSHT
506
1.618
7.404
0.745
0.678

GRGQVVSTHQNQQT
507
1.607
3.318
0.931
0.561

GSAQVSMVGQNQQT
508
1.601
1.332
0.500
0.285

GSSTLVTIGKNQQT
509
1.592
4.316
0.917
0.819

GFAHQASSGQNQQT
510
1.587
1.852
1.638
1.080

GSGQPVLSISNQQT
511
1.586
2.695
0.390
0.282

GSGQSHRSELNQQT
512
1.585
11.974
0.668
0.256

GSSVGSPIGQNQQT
513
1.584
3.574
1.059
0.706

GSGMPIRNVQNQQT
514
1.584
0.138
0.684
0.631

GSSTRVDSGQNQQT
515
1.584
2.774
0.704
0.660

GSGQNQQTAMRSTT
516
1.581
2.588
0.656
0.665

GSGQNQQHSSSHLT
517
1.581
2.782
1.091
0.859

GSRNGHAVGQNQQT
518
1.574
2.688
0.434
0.939

GLGAYQSSGQNQQT
519
1.574
0.696
1.407
0.688

GPGLSGHSGQNQQT
520
1.571
1.603
1.154
1.297

GSTGIVSSGQNQQT
521
1.570
0.927
2.141
1.046

GSRTTQVIGQNQQT
522
1.570
1.838
0.773
0.564

GSGLLHRAQQNQQT
523
1.569
0.724
1.583
0.646

GSGQNAQQAAAQQT
524
1.568
4.239
0.937
0.604

GSGQNQQSALRTQT
525
1.568
1.913
1.581
1.421

GSGFLSDTRQNQQT
526
1.566
45.953
0.473
0.575

GSGLLYHDQQNQQT
527
1.565
2.760
0.405
0.107

GSGQNQHYSLHKQT
528
1.563
3.399
1.485
1.273

GSGHSPLPQQNQQT
529
1.562
0.556
0.387
0.247

GNGHSMRPNQNQQT
530
1.560
2.341
0.693
0.376

GSGLKWSTLQNQQT
531
1.556
0.000
1.134
2.442

GSGQMGRQAVNQQT
532
1.554
1.529
0.535
0.411

GSGQNQQTSGVLTL
533
1.553
0.000
1.104
0.782

GSGQNQQALHNPHT
534
1.553
0.664
0.638
0.213

GSGQNQQVIPNSKT
535
1.548
1.036
0.844
0.376

GSPLQDRVGQNQQT
536
1.548
0.753
0.469
0.391

GSGQNQYSSTNPQT
537
1.542
2.251
0.544
0.535

GAMTVTISGQNQQT
538
1.542
6.249
0.443
0.257

GSGQNQQLQTLIRT
539
1.538
1.425
0.813
0.514

GSGLRQTSQQNQQT
540
1.537
2.067
0.978
0.705

GSGQNQQTGLRQQT
541
1.533
2.120
1.217
1.103

GSGQTRQMKDNQQT
542
1.530
11.079
0.841
0.214

GSGQNHGLQSGQQT
543
1.530
4.960
0.938
0.779

GSGQSHRQPENQQT
544
1.529
2.153
0.209
0.159

GSGQDRHIVQNQQT
545
1.527
11.068
0.285
0.162

GSGQNQQLPHSNLT
546
1.521
1.838
0.442
0.283

GSGQLSVPYDNQQT
547
1.521
0.000
0.622
0.111

GSGRNPQTQPLQQT
548
1.519
0.040
0.733
0.573

GSGQPYSTGLNQQT
549
1.519
1.403
0.612
0.376

GSGQNQQTHGGLRD
550
1.519
6.487
1.913
1.298

GAYGMVSSGQNQQT
551
1.518
3.469
0.732
0.773

GSGIQSSYSQNQQT
552
1.517
15.978
1.032
0.684

GPRLSDQSGQNQQT
553
1.511
0.364
0.640
0.579

GSGQNQQTHPSPCT
554
1.510
1.003
1.120
0.546

GSGQSFQMHTNQQT
555
1.504
9.770
0.503
0.325

GSGQNQQTGNPKHT
556
1.504
5.973
1.391
1.139

GFSSAVHSGQNQQT
557
1.502
1.234
0.218
0.210

GSGQNQQTSMSNAT
558
1.501
6.766
1.605
0.745

GSGQDMKQHHNQQT
559
1.501
1.638
0.358
0.239

GLRLSTPSGQNQQT
560
1.498
4.334
0.804
0.522

GSGQNQQTSVYMNT
561
1.498
0.613
0.640
0.983

GSGQNQYSQSSMQT
562
1.494
4.278
0.375
0.309

GSGQNQQSMADHTT
563
1.494
1.728
0.428
0.215

GWERSFVSGQNQQT
564
1.492
0.943
0.490
0.538

GLLAGKSSGQNQQT
565
1.491
2.981
0.999
0.946

GKSFVPQSGQNQQT
566
1.489
2.502
1.798
0.430

GSGQMQSAGSNQQT
567
1.482
0.116
1.034
1.128

GSDQNQRLTSSMQT
568
1.479
0.164
0.875
0.670

GESRAVLSGQNQQT
569
1.476
0.938
0.789
0.368

GSVFGVPSGQNQQT
570
1.474
1.248
0.685
0.213

GSGLPDRNLQNQQT
571
1.471
7.306
1.136
0.611

GSGTHNSAIQNQQT
572
1.469
0.570
0.762
0.574

GSGMIIASMQNQQT
573
1.469
6.722
1.135
0.415

GGITWTDSGQNQQT
574
1.462
4.535
1.472
0.468

GSGQNQQASGRQQT
575
1.458
3.179
0.943
0.991

GSGQNQQPHLKSLT
576
1.457
5.016
1.096
0.740

GPPQHMTSGQNQQT
577
1.457
1.547
0.509
0.677

GSGQNQQASLPSRT
578
1.456
0.389
0.930
0.673

GSGQIVSTQTNQQT
579
1.456
1.103
0.453
0.512

GSGKGHSAGQNQQT
580
1.453
0.936
1.035
1.173

GSGQNTRLQLGQQT
581
1.452
1.747
0.181
0.234

GSVGSRPVGQNQQT
582
1.442
11.363
1.182
0.716

GSSHTLALGQNQQT
583
1.441
7.071
0.851
0.406

GMYEYSQSGQNQQT
584
1.438
0.000
1.410
0.448

GNGQNQQHSILHGT
585
1.435
0.000
0.777
0.415

GSGYNQPHLQNQQT
586
1.435
4.512
0.711
0.395

GPLVNASSGQNQQT
587
1.434
5.239
0.831
0.343

GSGQNQQVLTTART
588
1.434
4.142
1.071
0.948

GSGQNQHSVHNDQT
589
1.428
0.000
0.521
0.515

GAGLIMHSGQNQQT
590
1.425
1.408
0.565
0.511

GMGRHSASGQNQQT
591
1.417
6.500
0.470
0.389

GSHSQSGHGQNQQT
592
1.413
1.240
0.696
0.318

GSSTTIVSGQNHQT
593
1.411
0.000
0.993
0.672

GRHLVTASGQNQQT
594
1.411
2.885
0.648
0.404

GSGQNQQHANLNQT
595
1.410
0.094
0.416
0.544

GSGSTHSKAQNQQT
596
1.410
0.515
0.921
0.801

GSGQNKQMLSGNTT
597
1.410
2.219
1.074
0.404

GSGQVHNPTQNQQT
598
1.410
2.488
1.021
0.542

GSGQNQQIPHVHQT
599
1.409
0.768
0.576
0.218

GSLHAGLSGQNQQT
600
1.408
1.739
1.286
0.936

GPAQHGTSGQNQQT
601
1.407
0.866
1.030
0.615

GEKAVTSSGQNQQT
602
1.402
0.998
0.558
0.327

GSGQNQQTMANGQR
603
1.394
0.216
1.169
1.230

GSGSPHSKDQNQQT
604
1.394
0.000
2.041
4.680

GSFSMGYGGQNQQT
605
1.393
18.476
1.908
1.030

GSGTHLVSLQNQQT
606
1.392
0.000
0.715
1.167

GSGQMQPHVQNQQT
607
1.389
9.381
0.387
0.153

GSGQNQQVAGLNNT
608
1.386
3.218
0.449
0.492

GSSQNQQHDMRLRT
609
1.386
2.645
0.669
0.552

GPASLPISGQNQQT
610
1.386
9.008
0.312
0.155

GSGQNQQPPLATRT
611
1.386
2.295
0.593
0.287

GSSRVPVSGQNQQT
612
1.385
13.191
0.870
0.485

GSGQNQQTNLGHTT
613
1.383
1.523
1.343
1.281

GSGQNQQLVSRVQT
614
1.381
1.195
0.656
0.466

GPNSYPVSGQKQQT
615
1.381
4.040
0.736
0.834

GHAHYQASGQNQQT
616
1.377
7.299
0.803
0.745

GSGQALLSTGNQQT
617
1.377
0.847
0.536
0.370

GSGQLPRQMTNQQT
618
1.376
3.550
0.400
0.562

GSGFPKSTEQNQQT
619
1.376
2.058
0.610
0.194

GSRETSLSGQNQQT
620
1.373
5.193
1.364
0.203

GSGQNQQGTGVSHT
621
1.371
4.295
1.417
0.749

GSRTVPVYGQNQQT
622
1.371
0.363
1.226
0.969

GSNAQSAHGQNQQT
623
1.371
0.888
0.976
0.245

GAFHLAASGQNQQT
624
1.369
18.165
0.994
0.775

GSGQYRSSSDNQQT
625
1.369
6.209
0.681
0.409

GSGQVYISTPNQQT
626
1.367
0.000
0.859
0.282

GSGVSTQLLQNQQT
627
1.367
2.467
0.928
0.509

GSGQLGLSVTNQQT
628
1.364
6.906
1.395
0.376

GSGSNMRLSQNQQT
629
1.363
0.588
0.962
0.730

GSGQNLHSGLPQQT
630
1.363
1.594
1.054
0.592

GSSHTLALGQNKQT
631
1.362
2.160
0.838
0.643

GSGQNQHSLPAHRT
632
1.361
0.700
0.911
0.742

GSGQNQGTVYPNQT
633
1.358
7.648
0.835
0.815

GSGQNQQPSLRQST
634
1.356
2.905
1.315
0.554

GSGQNARLKDNQQT
635
1.354
2.395
0.580
0.938

GHAGSTGSGQNQQT
636
1.352
2.829
1.332
1.233

GSGQALSSSGNQQT
637
1.351
6.860
0.894
0.931

GSGASESHRQNQQT
638
1.350
0.850
0.325
0.313

GVGVITSSGQNQQT
639
1.348
0.918
1.296
0.777

GSLYGQSLGQNQQT
640
1.348
11.248
0.894
0.843

GSGQMSDVHGNQQT
641
1.346
7.172
0.408
0.548

GSGQNQQHSSKATT
642
1.345
12.248
1.350
1.401

GSGQNQQTSVSQQT
643
1.342
1.614
1.030
0.913

GSGQKMWKLDNQQT
644
1.341
0.000
0.990
1.418

GSGQNVSMQVNQQT
645
1.341
0.000
0.357
0.251

GSGQNQRATLSNQT
646
1.339
1.084
0.947
0.723

GSGQASSKSANQQT
647
1.339
1.138
0.500
0.175

GSGKNQTPIPKGQT
648
1.339
5.077
1.306
1.154

GSGQNQQTRQEGST
649
1.339
0.000
0.645
0.718

GASSLATSGQNQQT
650
1.337
0.703
0.423
0.217

GSGQRGSLTENQQT
651
1.337
2.482
0.300
0.567

GSEQTRQRGQNQQT
652
1.333
2.172
0.574
0.815

GSGQNQQTLTASKE
653
1.333
1.152
0.981
1.172

GSGTSGKTGKNQQT
654
1.333
4.033
0.358
0.676

GQLVTFTSGQNQQT
655
1.331
11.282
0.819
0.294

GSGQNQQSANKILT
656
1.331
3.789
0.894
1.236

GSGQNQQHHSSHTT
657
1.328
2.158
0.957
0.452

GSGQNQKGMQPNQT
658
1.326
3.139
0.775
1.059

GSGQLVSGLYNQQT
659
1.325
0.000
0.842
0.733

GSSVGVPSGQNQQT
660
1.322
4.867
0.336
1.157

GSGQNQQWDSRRQT
661
1.321
0.531
1.059
0.825

GSEQTRQSGQNQQT
662
1.321
0.514
0.734
0.900

GSGIGSHIPQNQQT
663
1.319
0.173
0.822
0.597

GSGQNQRLHGVDQT
664
1.318
4.655
0.459
0.341

GEVSRVLSGQNQQT
665
1.318
0.437
1.150
0.440

GSGQNQQKVSPLLT
666
1.314
1.602
0.755
0.806

GSGLALERSQNQQT
667
1.311
0.486
0.618
0.096

GPDRIGSSGQNQQT
668
1.308
0.426
0.654
0.342

GSGQNQDHQNKQQT
669
1.308
1.470
0.510
0.761

GSGQNQQTALYNNT
670
1.307
0.862
0.660
0.726

GSGAVHLTAQNQQT
671
1.306
1.668
0.541
0.466

GSLVSTQSGQNQQT
672
1.305
1.293
1.282
0.650

GSGVSARMVQNQQT
673
1.299
0.624
0.870
0.697

GSGQTRMPLANQQT
674
1.296
0.790
0.447
0.273

GSGISSRNMQNQQT
675
1.291
6.328
1.671
0.560

GSGEKVHSGQNQQT
676
1.289
0.062
0.862
0.671

GSGQNQQKLSSMST
677
1.286
1.586
1.160
1.052

GSGQNQQTGQHMRV
678
1.286
4.161
1.839
1.635

GSGMIHTTAQNQQT
679
1.285
0.105
0.678
0.276

GSGQNWPALKGQQT
680
1.284
2.031
1.101
1.222

GASHMSISGQNQQT
681
1.284
0.462
0.404
0.374

GSDQNQQLGYSKQT
682
1.283
0.000
0.853
0.660

GIPSIRESGQNQQT
683
1.282
0.166
0.484
0.254

GSGIPSVKFQNQQT
684
1.281
0.061
0.364
0.561

GSGQNQQTSVSQNV
685
1.281
0.750
0.788
0.715

GSGQNQQIGESRMT
686
1.279
0.103
0.890
0.453

GSGSSSMSFQNQQT
687
1.279
0.540
0.466
0.095

GSGQKQERAVSKQT
688
1.277
0.000
1.174
0.732

GCTTRLNSGQNQQT
689
1.276
0.000
0.184
0.618

GSGQNQQIISTKIT
690
1.275
0.000
0.951
0.710

GSGQNQQKSLNGNT
691
1.275
8.573
0.586
0.851

GSGIPAPRLQNQQT
692
1.273
4.162
0.583
0.396

GSGQIRESMGNQQT
693
1.270
1.676
0.833
0.523

GSGQNSGVHFNQQT
694
1.268
0.587
0.871
0.377

GSGQNIGHSLPQQT
695
1.264
6.183
0.740
0.478

GSGERSISVQNQQT
696
1.264
1.619
0.598
0.173

GSGLKPNVLQNQQT
697
1.263
0.975
0.701
0.268

GSGQVAYAQGNQQT
698
1.259
1.309
0.734
0.313

GSGQSSYGSGNQQT
699
1.257
1.686
1.161
0.456

GSGQNQAMTHGDQT
700
1.257
1.878
0.357
0.259

GSGQNQALVSMGQT
701
1.255
1.876
0.987
0.560

GSGQNPSFMRGQQT
702
1.252
1.454
1.293
1.094

GSGQNQQSHLRINT
703
1.251
4.583
1.022
0.718

GYTRLETSGQNQQT
704
1.250
1.323
0.841
0.297

GSGQSYDMRGNQQT
705
1.248
0.567
0.588
0.368

GSRTTQDIGQNQQT
706
1.247
0.000
0.685
0.280

GSGHPYKAAQNQQT
707
1.246
0.000
0.872
0.507

GRLSNAHGGQNQQT
708
1.245
0.839
1.036
0.725

GSGQNQRAVLNDQT
709
1.242
3.023
0.556
0.259

GGSHTYGGGQNQQT
710
1.241
13.065
0.982
0.730

GSSVNSMIGQNQQT
711
1.239
0.000
0.976
0.580

GNSSMMGSGQNQQT
712
1.239
3.856
0.656
0.364

GNRDRPSSGQNQQT
713
1.239
3.947
0.298
0.178

GSGNMHASRQNQQT
714
1.238
3.878
0.782
0.687

GFIFPKVSGQNQQT
715
1.237
0.000
1.764
0.692

GSGQNQQLKNSTST
716
1.235
1.703
1.063
0.538

GSGQNQQSQYMPRT
717
1.234
0.401
0.549
0.520

GSGQRMADIGNQQT
718
1.233
2.539
0.352
0.427

GSGQNQSHYPSQQT
719
1.228
4.315
0.644
0.402

GSDGKMHRGQNQQT
720
1.227
0.000
1.826
0.776

GSGSVGFIGQNQQT
721
1.227
8.261
0.689
0.445

GLHGMTLSGQNQQT
722
1.226
3.552
0.470
0.338

GSDQSKRGDSNQQT
723
1.225
0.639
0.479
0.267

GSLFLATGGQNQQT
724
1.220
0.000
0.775
0.485

GSGQNQQPSAFSKT
725
1.220
4.906
1.309
0.754

GSGQLPQSGLNQQT
726
1.218
1.504
0.641
0.318

GSGSKQNALQNQQT
727
1.216
2.010
0.941
0.594

GSGQRRELSQNQQT
728
1.215
1.791
0.622
0.396

GSGQREPKASNQQT
729
1.214
2.793
0.399
0.520

GSGQNQQHPSTQQT
730
1.205
1.552
1.017
0.680

GSQSTLGLGQNQQT
731
1.204
3.246
0.594
0.400

GSGQNQQMPGLSST
732
1.204
1.887
0.234
0.181

GSGQNQQTVGGKNL
733
1.203
0.128
0.777
1.051

GSSREFHSGQNQQT
734
1.203
1.591
0.688
0.474

GSGQNQQTVPSNLV
735
1.201
0.791
0.434
0.281

GSGQNAYSSQAQQT
736
1.201
12.096
0.629
0.216

GSGQNKDHSTRRQT
737
1.197
0.000
0.384
0.477

GQLGSVGSGQDQQT
738
1.196
0.000
1.020
0.437

GSGQHAAPGHNQQT
739
1.195
5.999
0.600
0.199

GSGQNQQTSQSPPT
740
1.194
1.208
0.851
0.478

GSGNYRDHEQNQQT
741
1.193
7.389
0.287
0.222

GSGQHSNQHVNQQT
742
1.192
1.453
0.955
0.558

GSGQTARNGINQQT
743
1.192
2.030
1.002
0.472

GSGQNQQHYGSQGT
744
1.189
0.453
1.345
0.379

GSGSPQASRQNQQT
745
1.189
6.782
0.923
0.542

GSGFSHSMGKNQQT
746
1.188
9.809
1.381
0.611

GSGQSHSLETNQQT
747
1.188
1.319
0.520
0.363

GTEQTRQSGQNQQT
748
1.188
0.132
0.756
0.756

GSGRHLASVQNQQT
749
1.187
1.024
0.654
0.606

GLGSKNHSGQNQQT
750
1.187
5.046
0.825
0.224

GSGQNQQTSHFPSA
751
1.185
0.325
0.969
0.907

GSGQLSGTPQNQQT
752
1.185
1.382
1.025
0.643

GSGQNQQAPHKKET
753
1.180
0.598
0.994
0.689

GSGQNQQTLRGSLE
754
1.179
1.812
0.853
0.354

GSIAMTSHGQNQQT
755
1.178
1.435
0.551
0.438

GSPGVSPSGQNQQT
756
1.178
3.006
0.853
1.160

GSGQNQQTGSSSRV
757
1.176
0.580
0.995
1.128

GSGQHLPLLGNQQT
758
1.175
1.739
0.519
0.347

GSDHSHRGGQNQQT
759
1.174
0.504
0.818
0.331

GSGIVTKLGQNQQT
760
1.174
10.571
0.599
0.242

GSGQDVTKTGNQQT
761
1.173
4.523
0.531
0.035

GSGQNQQSHGRIGT
762
1.173
5.117
0.607
0.455

GSGQNQQINHRSPT
763
1.173
0.748
0.259
0.220

GSGDDSRVGQNQQT
764
1.172
0.191
0.466
0.156

GSGQSTLKRINQQT
765
1.168
13.442
0.534
1.184

GSGSQHSKAQNQQT
766
1.168
0.312
0.638
0.916

GSGQNQQHASSNNT
767
1.166
7.155
0.789
0.896

GSRTYQVSGQNQQT
768
1.164
1.853
0.638
0.641

GSGQNQGLLSSPQT
769
1.164
0.000
0.707
0.417

GSGGGLQHNQNQQT
770
1.163
4.098
1.137
0.778

GSGQNQQTTAATRM
771
1.163
3.925
0.947
1.005

GSGQNQRASILVQT
772
1.162
3.632
0.531
0.569

GSGQNLGLLGAQQT
773
1.161
1.458
0.524
0.226

GSLDLGRSGQNQQT
774
1.160
3.283
1.002
0.505

GNSQVKVSGQNQQT
775
1.158
4.930
1.422
0.728

GSSGSHQYGQNQQT
776
1.155
0.000
1.129
0.794

GSGQNQQQRDGTLT
777
1.152
0.387
0.760
0.730

GRGQHVSVANNQQT
778
1.152
1.896
1.032
0.589

GDSSSRISGQNQQT
779
1.151
3.787
0.916
0.348

GSGQNQQHSLSSQT
780
1.150
3.844
0.700
0.730

GSLMDVHRGQNQQT
781
1.150
0.387
1.009
0.238

GSIQYQSSGQNQQT
782
1.147
2.601
1.074
1.191

GLGSKNPSGQNQQT
783
1.147
1.629
1.184
0.424

GSGQLVLTLQNQQT
784
1.143
0.000
0.336
0.336

GSGQNQQTSQPLPG
785
1.141
0.080
0.748
0.530

GSGQNQQNLGKLNT
786
1.141
0.000
0.919
0.687

GTTAHQPSGQNQQT
787
1.138
0.211
0.726
0.275

GSGQNRAQIGTQQT
788
1.138
0.469
0.776
0.654

GSGQYVHVSSNQQT
789
1.137
1.803
0.739
0.366

GSGQNQQTAHAFNI
790
1.132
3.404
0.699
0.729

GSGQNQRTMVATQT
791
1.130
1.122
0.649
0.554

GSGQNPIRGAMQQT
792
1.126
1.327
1.296
0.427

GSGYVITGSQNQQT
793
1.125
6.271
0.971
0.248

GRGPKQSNIQNQQT
794
1.125
0.737
0.771
2.490

GSGQNQQTMLGKPC
795
1.125
0.047
1.090
0.992

GSGQNQQVGSTVRT
796
1.124
2.040
0.918
0.614

GNVTTQKSGQNQQT
797
1.122
2.546
1.215
0.922

GSGNPVSHLQNQQT
798
1.121
1.037
0.583
0.310

GSLSHMESGQNQQT
799
1.120
0.829
0.489
0.265

GRAPTNLSGQNQQT
800
1.118
0.687
0.757
0.169

GSGQNQQTVMTARA
801
1.117
1.535
0.995
0.843

GSGMPASRLQNQQT
802
1.117
1.689
0.790
0.372

GVVRNHQSGQNQQT
803
1.116
5.801
0.899
0.868

GSGQNQHSVQVRQT
804
1.116
1.909
0.782
0.916

GSGQNTGHLTMQQT
805
1.114
0.078
1.026
0.595

GSGQNQQYAGKILT
806
1.112
0.300
1.078
0.431

GSGNPHVRNQNQQT
807
1.112
0.873
0.732
0.755

GSGQNGGSSNRQQT
808
1.109
2.594
1.255
0.844

GSGQRLSQGVNHQT
809
1.108
3.394
0.931
1.141

GSGQNAHAKEGQQT
810
1.108
0.000
0.875
1.179

GSSPAPNSGQNQQT
811
1.106
2.229
0.719
0.368

GLAHKTSSGQNQQT
812
1.106
0.915
0.427
0.690

GSGQNQQTPGAHKT
813
1.105
3.827
0.957
0.277

GSGQNQQSLSGSFT
814
1.105
0.735
0.745
0.883

GSGQNQQSTGTSRT
815
1.103
4.054
1.209
0.935

GSGQNQQTVQSNLV
816
1.103
2.350
0.577
0.698

GSGQNQQLGSRQCT
817
1.102
0.183
0.987
0.407

GSGQNQYLRLELQT
818
1.101
0.000
0.416
0.839

GSGQNQQTSPRLQT
819
1.100
0.795
1.156
1.091

GSGQNQQTTSSNMT
820
1.099
0.569
0.638
0.698

GTASTYNSGQNQQT
821
1.099
2.560
0.250
0.625

GSGQNQQTMPQHKI
822
1.097
2.394
0.479
0.197

GSGQSHLHTGNQQT
823
1.096
2.584
0.721
0.295

GVKGVGHSGQNQQT
824
1.096
2.485
0.994
0.783

GSGKVTKQSQNQQT
825
1.095
0.000
0.928
1.035

GSGQNQQTALEKSL
826
1.092
0.000
0.625
0.702

GSGYKDTYGQNQQT
827
1.091
0.854
0.717
0.448

GSGQNQQSGTFLST
828
1.090
5.673
1.021
0.742

GSGQNTGQHMMQQT
829
1.090
1.058
1.147
0.917

GSGKNQQRPGLDQT
830
1.089
1.557
0.583
0.385

GSGQSREISLNQQT
831
1.088
6.954
0.594
0.282

GTPTSPSSGQNQQT
832
1.086
4.558
0.833
0.662

GKPAGGLSGQNQQT
833
1.085
2.805
0.708
0.739

GSGQNHRSADMQQT
834
1.084
12.001
0.417
0.212

GSGQNQQTLPSLSL
835
1.084
1.758
0.527
0.175

GSPYMGATGQNQQT
836
1.083
5.364
0.918
0.254

GSGHAKAVGQNQQT
837
1.081
4.357
0.703
0.824

GHMKGVTSGQNQQT
838
1.081
2.814
0.807
0.413

GSGQNQKILTLDQT
839
1.080
0.371
0.291
0.314

GSGQNQQTKVGHSA
840
1.079
1.256
0.669
1.019

GIARTTISGQNQQT
841
1.078
1.783
0.819
0.330

GSGQNQQTSVGFRT
842
1.077
3.737
0.648
0.534

GSGQNQQTMIANIR
843
1.076
0.000
0.379
0.458

GDMTRSSSGQNQQT
844
1.075
0.802
1.145
1.038

GSGHMSDLRQNQQT
845
1.073
4.291
0.555
0.328

GRGAVMASGQNQQT
846
1.072
0.923
0.783
0.605

GSGQNQQLSGKSVT
847
1.070
1.524
1.276
0.930

GSHTLVVSGQNQQT
848
1.069
1.535
0.671
0.748

GSGPWSAGLQNQQT
849
1.067
0.947
0.700
0.539

GSGQHSPHALNQQT
850
1.064
1.412
0.885
0.573

GSGQNQQPNSGSMT
851
1.064
0.925
0.588
0.339

GSGLAHLGGQNQQT
852
1.064
2.191
0.749
0.794

GSSVRYEPKQNQQT
853
1.063
1.564
0.450
0.501

GSGQNQQARPLELT
854
1.061
0.059
0.389
0.252

GSGQPRSTGINQQT
855
1.061
0.693
0.650
0.542

GSGQNQANWVKVQT
856
1.059
0.126
0.683
0.532

GSGHLFQSGQNQQT
857
1.057
0.615
0.751
0.386

GSGQNRGISISQQT
858
1.057
2.166
0.686
0.566

GSGTHYDNRQNQQT
859
1.054
0.072
0.612
0.486

GSGQNQQTSTTPLP
860
1.052
2.823
0.828
0.741

GSGQVHASQVNQKT
861
1.049
0.503
0.855
0.767

GSSGHRESGQNQQT
862
1.048
4.398
0.641
0.691

GLSAEKSSGQNQQT
863
1.047
7.203
0.629
0.303

GSGQEHRSLANQQT
864
1.046
0.000
0.507
0.344

GSGQTVVRIANQQT
865
1.046
4.156
0.661
0.390

GSGQNVSSVHRQQT
866
1.045
0.712
0.383
0.271

GSGASRMSIQNQQT
867
1.045
0.111
0.801
0.417

GVAFIGSSGQNQQT
868
1.043
0.000
0.744
0.648

GSGQNQQTVPTRQT
869
1.040
1.207
0.629
0.138

GSGQAAKSSQNQQT
870
1.036
0.681
0.778
0.737

GSGQNQQVAIRTST
871
1.035
2.447
0.963
0.370

GSVHMQNAGQNQQT
872
1.034
3.608
1.004
0.625

GSGMRQAGVQNQQT
873
1.032
0.811
0.736
0.775

GSGQNQQVGGKTVT
874
1.032
6.195
1.094
0.821

GVHDMRVSGQNQQT
875
1.032
8.083
1.171
0.818

GSGQHVSVANNQQT
876
1.029
5.734
0.974
0.577

GSAAMSVRGQNQQT
877
1.029
2.386
0.202
0.287

GVSRGGPSGQNQQT
878
1.028
1.611
0.750
0.591

GSGQMVHTIGNQQT
879
1.026
1.328
0.406
0.430

GRGGSMAETQNQQT
880
1.024
2.853
0.799
0.669

GSGHTNPTRQNQQT
881
1.021
0.688
0.726
0.807

GSGEAARYEQNQQT
882
1.020
0.000
0.107
0.125

GSGQNERHLVLQQT
883
1.019
5.354
0.416
0.150

GSGQNQQSKQQVLT
884
1.019
1.494
1.428
1.256

GSGQARAHRGNQQT
885
1.017
0.000
0.254
0.386

GSGQNQQPLDTSRT
886
1.015
0.775
0.491
0.376

GSGQNQQLANMVTT
887
1.014
1.739
1.253
0.987

GSGQMKDLHRNQQT
888
1.014
1.068
0.587
0.506

GSGQNQHLSSFVQT
889
1.013
0.110
1.090
0.364

GSGQNQQPSSRVTT
890
1.012
2.179
0.784
0.504

GSGQNQQLAITLGT
891
1.011
0.000
0.877
0.143

GSGQNQQTVGNPAT
892
1.008
3.014
0.856
0.395

GSGQNQGRAHPMQT
893
1.007
2.364
0.684
0.453

GSGQLIASVVNQQT
894
1.005
0.086
0.197
0.359

GSSVRSLVGQNQQT
895
1.004
3.840
0.412
0.608

GGAGSAHSGQNQQT
896
1.003
6.108
0.474
1.092

GSDQNQQTMSSTRT
897
1.003
2.428
1.306
0.835

GSGQNQQMAGAFRT
898
1.003
1.784
1.307
0.762

GSLGNLQRGQNQQT
899
1.003
0.895
0.947
0.385

GSGPSISHGQNQQT
900
1.000
0.000
0.614
0.665

GSGQNQQT
6406
1.000
1.000
1.000
1.000

GSGQNQQSSFNVQT
901
0.998
0.000
1.307
0.675

GSGQNQQTGQATHN
902
0.996
2.199
0.877
0.527

A second cross-species evolution screen was performed using an AAV capsid variant library with a modification in loop IV introduced as described above and passaging it once through NHPs (passage 1) and then subsequently injected it into two different strains of mice (passage 2), C571B1/6 and BALB/c. The fold-enrichment for each variant in the brain of each mouse species was calculated by systematic NGS enrichment analysis following RNA recovery and RT-PCR amplification. The fold enrichment values in the second passage in mice were compared to those fold enrichment values from the second pass that was performed in NHPs as described above. As shown in Table 10, when comparing the second pass fold enrichment values in the mice versus NHPs, 12 variants were identified that had a fold-enrichment value greater than 10 in all three animal groups. Further, 10 of these 12 variants comprised the SPH motif and a positive residue in one of the next three subsequent residues (Table 10).

TABLE 10

NGS fold-enrichment of AAV capsid variants from a second passage (P2) in

NHPs or mice (C57Bl/6 or BALB/c) following a first passage in NHPs

SEQ

Fold enrichment

ID
Fold enrichment over
Fold enrichment over
over AAV9 in

Peptide Sequence
NO:
AAV9 in NHP P2
AAV9 in BALB/c P2
C57Bl/6 P2

VSGSPHSKAQNQQT
903
99.76
92.99
34.29

CSGSPHSKAQNQQT
904
85.1
66.74
22.19

GSGSPHSKAQNQQT
200
56.33
44.58
14.48

GSGSPHRKAQNQQT
905
46.39
42.47
14.11

GRGSPHSKAQNQQT
906
43.68
59.65
28.13

GHDSPHKSGQNQQT
201
33.96
59.14
27.15

GSGSPHSKAKNQQT
208
31.27
41.51
14

GSGSPHSKAQNKQT
907
29.52
44.1
13.69

GSGSPHSKAQTQQT
908
24.27
41.75
18

GQDSPHKSGQNQQT
204
22.7
32.37
16.02

GSGSTHASRQNQQT
909
11.04
23.71
10.67

GHDSQHKSGQNQQT
404
10.36
21.3
13.55

Following the second passage in mice, a synthetic library was generated using those variants that demonstrated a fold-change in enrichment relative to wild-type AAV9 that was above 10 in the brain of either strain of mice, as measured by systematic NGS enrichment analysis following RNA recovery and RT-PCR amplification. There were approximately 500 variants in this synthetic library. This synthetic library was then injected back into both strains of mice (C57B1/6 and BALB/c; passage 3). RNA was recovered from the mouse brains, RT-PCR amplification was performed, and fold-enrichment relative to wild-type AAV9 was calculated by NGS analysis, which is provided in Table 11. As shown in Table 11, the variants with the greatest fold-enrichment in the brain in each strain, were highly correlated across strains (R²=0.8458).

TABLE 11

NGS fold-enrichment of AAV capsid variants in the brain from a third passage

(P3) in mice (C57Bl/6 or BALB/c) following a first and second passage in mice

Fold enrichment
Fold enrichment

over AAV9 in
over AAV9 in

Peptide Sequence
SEQ ID NO:
BALB/c
C57Bl/6
Average

GSGSPHKYGQNQQT
910
150.445
103.488
126.966

GSGSPHKFGQNQQT
911
73.364
60.304
66.834

GHDSPHKSGQNQQT
201
82.460
51.125
66.792

GSGSPHSKAQNQQT
200
60.312
65.853
63.083

VSGSPHKFGQNQQT
912
60.186
59.142
59.664

GSGSPHSKAQNHQT
913
63.486
51.647
57.566

VSGSPHSKAQNQQT
903
73.555
37.429
55.492

GQDSPHKSGQNQQT
204
63.898
43.752
53.825

GSGSPHSKAQHQQT
914
45.309
45.600
45.454

GSGSPHKTYQNQQT
915
50.283
35.460
42.871

GSGSPHSKAQTQQT
908
43.120
39.098
41.109

VSGSPHASRQNQQT
916
46.572
32.480
39.526

GSGSPHSKAQNKQT
907
39.848
35.596
37.722

GSGSPHKFGKNQQT
917
31.948
34.899
33.423

GSGSPHASRQNQHT
918
28.145
30.928
29.537

GSHSPHKSGQNQQT
919
22.948
35.412
29.180

GSGQNQQRRMSPST
920
4.576
53.520
29.048

GSGSPHASRQNQQT
205
28.866
29.139
29.003

GSGSPHSKPQNQQT
921
26.958
28.599
27.779

GSGSPHKFGQKQQT
922
39.597
14.927
27.262

VSGSPHGARQNQQT
923
30.985
22.634
26.810

GSGSPHSKAQKQQT
924
25.052
27.459
26.256

GHSSPHRSGQNQQT
212
16.982
35.081
26.032

GSGSPHSKAKNQQT
208
21.069
25.711
23.390

GSHSPHKRGQNQQT
925
24.054
20.262
22.158

GRGSPHSKAQNQQT
906
20.939
22.720
21.830

GQSSPHRSGQNQQT
229
9.916
26.608
18.262

GSGQNRQRLKGLET
926
3.937
31.022
17.480

GSGSPHKLGQNQQT
927
18.905
14.732
16.818

GSGSPHKTSKNQQT
928
14.654
17.606
16.130

GSGSPHKIGQNQQT
929
16.999
14.794
15.897

GSGSPHKKNQNQQT
209
25.633
5.605
15.619

GSGSPHASRQNKQT
206
10.738
20.347
15.542

GSGSPHTRGQNQQT
214
16.899
13.869
15.384

GSGQDSPHVRNQQT
930
15.340
14.646
14.993

GSGSPHKTSQNQQT
931
20.428
8.818
14.623

GSGSPHASRKNQQT
932
13.799
12.749
13.274

GSGSPHASRQKQQT
211
13.624
11.188
12.406

GSHSPHKSGQKQQT
933
6.700
17.736
12.218

GSGSPHKTSQKQQT
934
12.621
11.720
12.170

GSGSPHVRGQNKQT
935
13.174
11.017
12.095

GSGSPHKTTQNQQT
936
9.722
13.381
11.552

CSGSPHSKAQNQQT
904
11.772
9.447
10.610

GSGPVRALRQNQQT
937
3.369
17.431
10.400

GSGSPHVRGQKQQT
938
7.573
12.498
10.036

GSGSPHRKAQNQQT
905
12.308
7.349
9.828

GRGSPHASRQNQQT
318
11.903
6.780
9.342

CSGSPHKTSQNQQT
939
11.167
6.631
8.899

CSHSPHKSGQNQQT
940
11.356
6.304
8.830

GSGSPHSKDQNQQT
604
3.492
10.236
6.864

Taken together, these results demonstrate that after 3 rounds of screening of this AAV9 variant library with loop IV modifications in NHP and mice, many AAV capsid variants outperformed the wild-type AAV9, for example, in penetration of the blood brain barrier (BBB) and spinal cord expression. These capsid variants were able to cross-species, evidenced by expression and tropism in the NHP brain/spinal cord as well as in the brain of two different mouse species.

Example 2. Individual Capsid Characterization in Mice

The goal of these experiments was to determine the transduction level, tropism, ability to cross the blood brain barrier, and overall spatial distribution in the central nervous system (CNS) of 2 capsid variants selected from the study described in Example 1 relative to AAV9 following intravenous injection in mice. The 2 capsid variants were TTM-001 (SEQ ID NO: 981 (amino acid) and 983 (DNA), comprising SEQ ID NO: 941) and TTM-002 (SEQ ID NO: 982 (amino acid) and 984 (DNA), comprising SEQ ID NO: 2), as outlined in Table 3 above. The amino acid and DNA sequences of TTM-001 and TTM-002 are provided, e.g., in Tables 4 and 5, respectively.

AAV particles were generated with each of these capsid variants encapsulating a luciferase-EGFP transgene driven by a CMV/chicken beta actin promoter in a single stranded viral genome. Each capsid variant and AAV9 control were tested by intravenously administering by tail vein injection, the AAV particle formulation at 5el 1 VG/dose (2.5E13 vg/kg) to three female BALB/c mice. The in-life period was 28 days and then various CNS and peripheral tissues were collected for measuring transgene mRNA, transgene protein, and viral DNA (biodistribution).

At 28 days post-injection of the AAV particles encapsulated in the TTM-001 capsid variant (AAV_TTM-001), mice were injected with luciferin and their brains were harvested for IVIS imaging. Robust luciferase signal was observed in mice injected with AAV particles encapsulated in the TTM-001 capsid variant, and this was greatly increased relative to AAV particles encapsulated in the wild-type AAV9 control capsid.

The brains isolated from mice injected with the AAV particles encapsulated in the TTM-001 capsid variant (AAV_TTM-001) or the TTM-002 capsid variant (AAV_TTM-002) were assayed by qPCR for the presence of transgene RNA as a measure of transgene expression, and the presence of viral DNA as a measure of viral genome levels. Data were provided as fold over AAV9 (Table 12). As shown in Table 12, when compared to the wild-type AAV9 capsid control, TTM-001 and TTM-002 demonstrated a 30-fold and 66-fold increase, respectively, in transgene mRNA levels and expression in the brain, indicative of enhanced payload delivery. This correlated with a 32-fold (TTM-001) and 47-fold (TTM-002) increase, respectively, in viral genome (DNA) concentrations in the brain relative to the AAV9 capsid control, which is indicative of enhanced CNS tropism and transduction (Table 12).

TABLE 12

Transgene mRNA and viral genome levels

(DNA) in mice relative to the AAV9 control

Measure
Tissue
AAV9
TTM-001
TTM-002

mRNA
Brain
1.0
30.4503
66.2161

(transgene expression)

DNA
Brain
1.0
32.0315
47.2810

(viral genome quantification)

mRNA
Liver
1.0
1.2356
0.2016

(transgene expression)

DNA
Liver
1.0
0.4802
0.0277

(viral genome quantification)

The brain tissues and spinal cords of the mice were also subjected to anti-GFP immunohistochemistry staining to evaluate overall CNS tropism and biodistribution. Immunohistochemical staining correlated with the qPCR analysis, as TTM-001 and TTM-002 showed significantly stronger staining and payload expression in the brain and spinal cord, as compared to the AAV9 control. More specifically, TTM-001 and TTM-02 demonstrated localization and strong payload expression and transduction in the mid-brain region, with increased staining observed in the hippocampus and thalamus, as well as in the brain stem, compared to AAV9. Less staining was observed in the cortical regions of the brain compared to the midbrain. However, staining in these cortical regions was stronger for TTM-001 and TTM-002 compared to the AAV9 control. It also appeared that the TTM-001 and TTM-002 capsid variants were able to transduce non-neuronal cells, including glial cells and oligodendrocytes. With respect to the spinal cord, staining and payload expression for TTM-01 and TTM-002 were localized to the ventral horns of the grey matter.

Peripheral tissues were also isolated from the mice intravenously injected with the AAV particles encapsulated in the TTM-001 capsid variant or the TTM-002 capsid variant for analysis by qPCR and/or GFP immunohistochemical staining. Transgene mRNA levels and viral genome DNA levels were quantified in the liver by qPCR and the fold over AAV9 was calculated for each capsid variant (Table 12). TTM-001 resulted in similar levels of payload expression (mRNA levels) as compared to wild-type AAV9, but only half as much viral genome DNA was quantified in the liver compared to AAV9. TTM-002 demonstrated greatly reduced mRNA and viral genome DNA levels in the liver compared to AAV9. GFP immunohistochemical staining of the spleen, heart, skeletal muscle, kidneys, and lungs of mice injected with AAV particles encapsulated in the TTM-001 capsid variant or the TTM-002 capsid variant showed similar levels of payload expression as compared to those mice injected with AAV particles encapsulated in the wild-type AAV9 control capsid.

Taken together, these data demonstrate that TTM-001 and TTM-002 are enhanced CNS tropic capsids in mice that can infect non-neuronal cells. Additionally, these capsid variants were able to successfully penetrate the blood brain barrier following intravenous injection.

Example 3. Maturation of TTM-001 and TTM-002 Capsid in Mice

This Example describes maturation of the TTM-001 (SEQ ID NO: 981 (amino acid) and 983 (DNA), comprising SEQ ID NO: 941) and TTM-002 (SEQ ID NO: 982 (amino acid) and 984 (DNA), comprising SEQ ID NO: 2) capsid variants to further enhance their transduction and biodistribution in the central nervous system and evolve the AAV capsid variants to provide further cross-species compatibility. Two approaches were used to mature the TTM-001 and TTM-002 capsid sequences in order to randomize and mutate within and around the peptide insert comprised within loop IV of the capsid variant. As many of the AAV capsid variants that demonstrated the greatest fold-enrichment in the NHP brain relative wild-type AAV9 comprised an SPH motif in the same position (e.g., immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138) (see Example 1), the SPH motif was not mutated in either approach to mature the TTM-001 and TTM-002 capsid variants. In the first maturation approach, sets of three contiguous amino acids were randomized across the mutagenesis region in the TTM-001 and TTM-002 sequences, which spanned from position 450 to position 466, numbered according to SEQ ID NO: 981 and 982. In the second maturation approach, mutagenic primers were used to introduce point mutations at a low frequency, scattered across the mutagenesis region in the TTM-001 and TTM-002 sequences ranging from position 449 to position 466, numbered according to SEQ ID NO: 981 and 982. AAV capsid variants arising from each maturation approach for TTM-001 were pooled together and AAV capsid variants arising from each maturation approach for TTM-002 were also pooled together, for subsequent testing and characterization in mice.

The library of pooled matured AAV capsid variants generated from TTM-001 or library of pooled matured AAV capsid variants generated from the TTM-002 matured AAV capsid variant each were intravenously injected into the tail vein of three female CD-1 Outbred mice (Charles River) at a dose of 1.0×10¹²VG/dose. After 14-days in life, the brains of the mice were isolated and RNA was extracted. Following RNA recovery and RT-PCR amplification, a systematic NGS enrichment analysis was performed to calculate the fold enrichment ratio relative to the corresponding TTM-001 or TTM-002 control, and the peptides comprised within the variants were identified. The data for the TTM-001 matured capsid variants is provided in Table 13 and the data for the TTM-002 matured capsid variants is provided in Table 14.

As shown in Table 13, approximately 714 TTM-001 matured capsid variants demonstrated at least a 2-fold increase in expression relative to the non-matured TTM-001 control, and several variants demonstrated greater than a four-fold enrichment relative to the non-matured TTM-001 control. Also, across the peptides comprised within the TTM-001 matured capsid variants with the greatest fold-enrichment relative to the non-matured TTM-001 capsid in the brain, it was observed that the modifications in the variant sequences appeared in the region C-terminal to the SPH motif present within the capsid variant. This indicates that modifications that appeared to improve TTM-001 capsid tropism in the CNS of mice were skewed to the C-terminal portion of the peptide insertion in loop IV of the sequence. Additionally, a number of these C-terminal modifications were the incorporation of an arginine (R) or leucine (L) residue.

TABLE 13

NGS fold-enrichment of TTM-001 matured AAV capsid

variants in the brain of CD-1 Outbred mice

Fold

SEQ
enrichment

ID
over TTM-

Peptide Sequence
NO:
001

KTINGSGSPHSLLWNQQT
1008
7.983

KTINGSGSPHSKAQYYVT
1009
6.283

KTINGSGSPHSKLRRQQT
1010
6.231

KTINGSGSPHSIWQNQQT
1011
5.883

KTINGSSSPHCTAQNQQT
1012
5.607

KTINGSGSPHSKAGCGQT
1013
5.341

KSMNGSGSPHSRAQNQQT
1014
5.145

KTINGSGSPHSKRLRQQT
1015
5.034

KTINGSGSPHSLRRNQQT
1016
4.985

KTINGSGSPHSRGRNQQT
1017
4.961

KTINGSGSPHSEIVNQQT
1018
4.931

KTINGSGSPHSSRRNQQT
1019
4.920

KTINGSGSPHCLLQNQQT
1020
4.898

KTIMRVGSPHSKAQNQQT
1021
4.875

KTINGSGSPHSKAFRLQT
1022
4.849

KTINGSGSPHCLAQNQQT
1023
4.847

KTINGSCSPHRKAQNQQT
1024
4.801

KTINGSGSPHFLRQNQQT
1025
4.777

KTINGSGSPHSLRFNQQT
1026
4.765

KTINGSGSPHSYLRNQQT
1027
4.566

KTINGSGSPHCSLQNQQT
1028
4.540

KTINGSGSPHVLWQNQQT
1029
4.533

KTINGSGSPHSKWLLQQT
1030
4.521

KTINGSGSPHSLWSNQQT
1031
4.467

KTINGSGSPHSKRRLQQT
1032
4.451

KTINGSGSPHSVYLNQQT
1033
4.426

KTINGSGSPHSLWLNQQT
1034
4.412

KTINGSGSPHSKAQRKLT
1035
4.339

KTINGSGSPHSKALRRQT
1036
4.330

KTINGSGSPHSKAQRLRT
1037
4.322

KYLSGSGSPHSKAQNQQT
1038
4.264

KTINGSGSPHSKAQRRLT
1039
4.227

KTINGSGSPHSKARRQQT
1040
4.218

KTINGSGSPHSKARRLQT
1041
4.210

KTINGSGSPHSKSRRQQT
1042
4.175

KTINGLLSPHWKAQNQQT
1043
4.173

KTINGSGSPHSKARLRQT
1044
4.155

KTINGSGSPHSKASKRQT
1045
4.117

KTINGSGSPHVRRQNQQT
1046
4.114

KTINGSGSPHSKAQLYRT
1047
4.108

KGLSGSGSPHSKAQNQQT
1048
4.056

KTINGSGSPHSLFRNQQT
1049
4.037

KTINGSGSPHSKAQLTVT
1050
4.026

KTINGSRSPHTRAQNQQT
1051
3.989

KTINGSGSPHSKAKLRQT
1052
3.976

KTINGSGSPHSKLIRQQT
1053
3.968

KTINGSGSPHSKALRFQT
1054
3.894

KTINGSGSPHSKRTFQQT
1055
3.879

KTINGSGSPHSKAQKRLT
1056
3.872

KTINGSGSPHLWSQNQQT
1057
3.857

KTINGSGSPHLLWQNQQT
1058
3.855

KTINGSGSPHSRLRNQQT
1059
3.851

KTINGSGSPHSKRAAQQT
1060
3.838

KTINGSGSPHSKRSWQQT
1061
3.838

KTINGSGSPHSKAQLRRT
1062
3.825

KTINGSGSPHYLVQNQQT
1063
3.819

KTINGSGSPHSKLFRQQT
1064
3.806

KTINGSGSPHSKRAMQQT
1065
3.801

KTINGSGSPHSKTLRQQT
1066
3.788

KTINGSGSPHSRSRNQQT
1067
3.784

KTINGSGSPHRRRQNQQT
1068
3.754

KTINGSGSPHSKTCLQQT
1069
3.717

KTINGSGSPHSKSRWQQT
1070
3.698

KTINGSGSPHRFRQNQQT
1071
3.698

KTINGLRSPHRKAQNQQT
1072
3.676

KTRSRSGSPHSKAQNQQT
1073
3.669

KTINGSGSPHSKAQLVVT
1074
3.654

KTINGSGSPHSRKLNQQT
1075
3.646

KTINGSGSPHSLLCNQQT
1076
3.644

KTINGKRSPHSKAQNQQT
1077
3.611

KTINGSRSPHLFAQNQQT
1078
3.601

KSINGSGSPHSKAHDQQT
1079
3.592

KTINGSGSPHSKAQRSRT
1080
3.585

KTINGSGSPHSTWLNQQT
1081
3.583

KTINGSGSPHSKASRRQT
1082
3.577

KTINGSGSPHSKRSMQQT
1083
3.561

KTINGSGSPHSCLQNQQT
1084
3.559

KTINGSGSPHSKRLWQQT
1085
3.529

KTINGSGSPHSWLSNQQT
1086
3.495

KTINGSGSPHLRRQNQQT
1087
3.493

KTINGSGSPHSKARRSQT
1088
3.493

KTINGSGSPHSKHLRQQT
1089
3.438

KTINGSGSPHSCSQNQQT
1090
3.428

KTINGSGSPHSKSFRQQT
1091
3.426

KTINGSGSPHLCLQNQQT
1092
3.425

KTINGSGSPHSKAQTSRT
1093
3.421

KTINGSGSPHSLCSNQQT
1094
3.413

KTINGSRSPHLRAQNQQT
1095
3.410

KTINGSGSPHSKAQVSKT
1096
3.406

KTINGSGSPHSKAQRHVT
1097
3.404

KTINGSSSPHLCAQNQQT
1098
3.402

KTINGSGSPHSFLRNQQT
1099
3.384

KTINGSGSPHSFVLNQQT
1100
3.382

KTINGSGSPHSKMRAQQT
1101
3.382

KTINGSGSPHRPRQNQQT
1102
3.380

KTINGSGSPHSKCLLQQT
1103
3.374

KTINGSGSPHSKAQSRRT
1104
3.372

KTINGSGSPHSRWQNQQT
1105
3.372

KYSVGSGSPHSKAQNQQT
1106
3.365

KTINGSGSPHSKRFLQQT
1107
3.359

KTINGSGSPHSLFLNQQT
1108
3.358

KTINGSGSPHSKAYLRQT
1109
3.356

KTINGSGSPHSKRNGQQT
1110
3.350

KTINGSGSPHTRRQNQQT
1111
3.350

KTINGSGSPHSKPRLQQT
1112
3.337

KTINFLRSPHSKAQNQQT
1113
3.331

KTINGSGSPHLLCQNQQT
1114
3.328

KTINGSGSPHSKARIVQT
1115
3.287

KTINGSKSPHFKAQNQQT
1116
3.285

KTINGSGSPHSKAQIRLT
1117
3.279

KTINGSSSPHWVAQNQQT
1118
3.277

KTINGSGSPHSKATRRQT
1119
3.277

KTINGSLSPHSCAQNQQT
1120
3.268

KTINGSGSPHSLYLNQQT
1121
3.264

KTINGSGSPHSKVGRQQT
1122
3.255

KTINGSGSPHSRRLNQQT
1123
3.251

KTINGSGSPHSKAQHSRT
1124
3.227

KTINGSGSPHSKAFPRQT
1125
3.220

KTINGSPSPHRRAQNQQT
1126
3.216

KTINGSGSPHSKRNLQQT
1127
3.210

KTINGSGSPHSKPTRQQT
1128
3.201

KTINGSGSPHSKLWLQQT
1129
3.199

KTINGSGSPHWLAQNQQT
1130
3.192

KTINGSGSPHRTRQNQQT
1131
3.190

KTINGSGSPHSKLNKQQT
1132
3.181

KTINGSGSPHSSLWNQQT
1133
3.179

KTINGSGSPHSKAQITLT
1134
3.177

KTINGSGSPHSKFLFQQT
1135
3.173

KTINGSGSPHSKRTPQQT
1136
3.169

KTINGSGSPHSKAQNSRR
1137
3.168

KTINGSGSPHSRLKNQQT
1138
3.156

KTINGSGSPHSCLLNQQT
1139
3.127

KTINGSGSPHTLYQNQQT
1140
3.117

KTINGSGSPHSKYPSQQT
1141
3.114

KTINGSGSPHSKLRNQQT
1142
3.112

KTINGSGSPHLNWQNQQT
1143
3.112

KTINGVVSPHRKAQNQQT
1144
3.106

KTINGSGSPHSYRPNQQT
1145
3.095

KTINGSTSPHRRAQNQQT
1146
3.089

KTINGSCSPHPLAQNQQT
1147
3.086

KTINGSGSPHSKAFARQT
1148
3.082

KTINGSGSPHSKALRYQT
1149
3.076

KTINGSKSPHRLAQNQQT
1150
3.073

KTINMRVSPHSKAQNQQT
1151
3.073

KTINGSGSPHMYLQNQQT
1152
3.061

KTINGSGSPHSKLARQQT
1153
3.054

KTINGSGSPHSKARPYQT
1154
3.050

KTINGSGSPHSKRVPQQT
1155
3.048

KTINGSGSPHLSWQNQQT
1156
3.047

KTINGRSSPHGKAQNQQT
1157
3.035

KTINGSGSPHLWTQNQQT
1158
3.034

KTINGLLSPHRKAQNQQT
1159
3.026

KTINGSGSPHRLRQNQQT
1160
2.998

KTINGSCSPHSGAQNQQT
1161
2.994

KTINGSGSPHSKAQRRST
1162
2.993

KTINGSGSPHSKLCSQQT
1163
2.989

KTINGSGSPHSKAQLLKT
1164
2.985

KTINGRKSPHSKAQNQQT
1165
2.985

KTINGSGSPHLLYQNQQT
1166
2.983

KTINGSGSPHSKLLRQQT
1167
2.981

KTINGSGSPHSLRHNQQT
1168
2.980

KTINGSGSPHSSKRNQQT
1169
2.978

KTINGSGSPHSKARSRQT
1170
2.972

KTINGRSSPHRKAQNQQT
1171
2.965

KTINGSKSPHRTAQNQQT
1172
2.950

KTINGMRSPHVKAQNQQT
1173
2.937

KTINGSGSPHSKRMSQQT
1174
2.931

KTINGSGSPHSKVPKQQT
1175
2.924

KTINLIRSPHSKAQNQQT
1176
2.920

KTINGSGSPHPFLQNQQT
1177
2.916

KTINGSGSPHSKARLWQT
1178
2.914

KTINGSGSPHSRTRNQQT
1179
2.912

KTINGSGSPHSKRSNQQT
1180
2.886

KTINGSLSPHSWAQNQQT
1181
2.885

KTINGSRSPHYKAQNQQT
1182
2.879

KTINRHSSPHSKAQNQQT
1183
2.877

KTINGSGSPHSKRRNQQT
1184
2.877

KTINGSGSPHSKAKHLQT
1185
2.870

KTINGSGSPHSKRTYQQT
1186
2.870

KTINGVLSPHRKAQNQQT
1187
2.868

KTINGSGSPHSFITNQQT
1188
2.868

KTINGSGSPHSTRLNQQT
1189
2.860

KTINGSGSPHSKRTSQQT
1190
2.857

KTINGSGSPHSRRSNQQT
1191
2.851

KTINGHLSPHRKAQNQQT
1192
2.851

KTINGSGSPHSKAQFSRT
1193
2.847

KTINGSGSPHSKAQTFRT
1194
2.847

KTINGSGSPHSKPLRQQT
1195
2.844

KTINGSGSPHSKASCRQT
1196
2.840

KTINGSGSPHSKILWQQT
1197
2.838

KTINGSGSPHSKALKRQT
1198
2.836

KTINGSGSPHSKAHRSQT
1199
2.819

KTINGSGSPHSMLYNQQT
1200
2.808

KTINGSGSPHSKCTLQQT
1201
2.808

KTINGSGSPHSKAQNRMR
1202
2.804

KTINGSGSPHSKLVRQQT
1203
2.801

KTINGSGSPHSKRILQQT
1204
2.801

KTINGSGSPHSKAQWLRT
1205
2.795

KTINGSGSPHSLTCNQQT
1206
2.795

KTINGIRSPHTKAQNQQT
1207
2.793

KTINGSGSPHSKAQRWLT
1208
2.788

KTINGSGSPHSKAQLSIT
1209
2.784

KTINGSGSPHIYRQNQQT
1210
2.782

KTINGSGSPHSLRSNQQT
1211
2.778

KTINGSGSPHSKVKPQQT
1212
2.778

KTINGSGSPHSKATRHQT
1213
2.777

KTINGSLSPHLCAQNQQT
1214
2.775

KTINGSGSPHSKACASQT
1215
2.775

KWSPGSGSPHSKAQNQQT
1216
2.764

KTINGYLSPHRKAQNQQT
1217
2.762

KTINGSGSPHSKVIRQQT
1218
2.760

KTINGSGSPHFLLQNQQT
1219
2.758

KTINGSGSPHSKARSKQT
1220
2.758

KTINGVPSPHWKAQNQQT
1221
2.758

KTINGSGSPHSKATRNQT
1222
2.756

KTINGSGSPHSKACSAQT
1223
2.754

KTINGSGSPHSKARYVQT
1224
2.749

KTINGSRSPHARAQNQQT
1225
2.745

KTINGSGSPHSKAQHLRT
1226
2.741

KTINGSGSPHSKAKSRQT
1227
2.739

KTINGSGSPHSKIGRQQT
1228
2.739

KTINGLASPHRKAQNQQT
1229
2.737

KTINGSGSPHSKARTRQT
1230
2.737

KTINGSGSPHSKSIRQQT
1231
2.728

KTINGSGSPHSKRLYQQT
1232
2.721

KTINGLPSPHRKAQNQQT
1233
2.719

KTINGSLSPHRRAQNQQT
1234
2.717

KTINGKTSPHGKAQNQQT
1235
2.717

KTINGSRSPHRLAQNQQT
1236
2.698

KTINGSGSPHSLTWNQQT
1237
2.698

KTINGSKSPHRKAQNQQT
1238
2.696

KTINGSGSPHSKAQLRKT
1239
2.689

KTINGSGSPHSKSRHQQT
1240
2.685

KTINRRLSPHSKAQNQQT
1241
2.678

KTINGSGSPHSRRVNQQT
1242
2.676

KTINGSGSPHSHWQNQQT
1243
2.676

KTTHCSGSPHSKAQNQQT
1244
2.672

KTINGSGSPHSWLQNQQT
1245
2.665

KTINGSTSPHYLAQNQQT
1246
2.665

KTINGLTSPHRKAQNQQT
1247
2.663

KTINGSGSPHSKRLLQQT
1248
2.659

KTINGSGSPHSKLCVQQT
1249
2.659

KTINGFLSPHRKAQNQQT
1250
2.654

KTINGSGSPHSKMRPQQT
1251
2.652

KTINGSGSPHSKQTRQQT
1252
2.650

KTINGSGSPHSYLINQQT
1253
2.650

KTINGSGSPHSKALRSQT
1254
2.648

KTINGMLSPHRKAQNQQT
1255
2.646

KTINGSGSPHSKCLTQQT
1256
2.644

KTINGSGSPHSKAQLTLT
1257
2.641

KTINGHSSPHRKAQNQQT
1258
2.639

KTINGSGSPHLTWQNQQT
1259
2.637

KTINGSGSPHSKAQYCLT
1260
2.628

KTINGSGSPHSFLVNQQT
1261
2.624

KTINMSRSPHSKAQNQQT
1262
2.622

KTINGSGSPHSKAQLHRT
1263
2.618

KTINGSGSPHLYMQNQQT
1264
2.615

KTINGSRSPHRRAQNQQT
1265
2.615

KTINGSGSPHSKAQNRRS
1266
2.613

KTINLRFSPHSKAQNQQT
1267
2.611

KTINGSGSPHSKAQRLWT
1268
2.611

KTINGSGSPHSKGRAQQT
1269
2.607

KTINGSGSPHSLSCNQQT
1270
2.605

KTINGLVSPHCKAQNQQT
1271
2.605

KTINGSSSPHLWAQNQQT
1272
2.605

KTINGSGSPHSKAHRLQT
1273
2.603

KTINGSGSPHPYAQNQQT
1274
2.598

KTINGSGSPHSTRPNQQT
1275
2.598

KTINGRSSPHPKAQNQQT
1276
2.596

KTINGSGSPHSKAQSWRT
1277
2.596

KTINGQRSPHVKAQNQQT
1278
2.596

KTINGSGSPHSKAQFVRT
1279
2.596

KTINGSGSPHSKCLNQQT
1280
2.594

KTINGSGSPHSSLCNQQT
1281
2.592

KTINGQRSPHSKAQNQQT
1282
2.590

KTINGSGSPHSLSWNQQT
1283
2.588

KTINGSGSPHSSRKNQQT
1284
2.585

KTINGSGSPHSKRTLQQT
1285
2.583

KTINGSLSPHCLAQNQQT
1286
2.577

KTINGSGSPHSKAQSSRT
1287
2.575

KTINGSGSPHLKRQNQQT
1288
2.564

KTINGSGSPHSKARMGQT
1289
2.564

KTINGSGSPHSKAQVKLT
1290
2.564

KTINGSGSPHSKLPRQQT
1291
2.562

KTINGSGSPHSKLCLQQT
1292
2.562

KTINGSGSPHSPLWNQQT
1293
2.562

KTINGSVSPHSWAQNQQT
1294
2.562

KTIRSKGSPHSKAQNQQT
1295
2.559

KTINGSRSPHSWAQNQQT
1296
2.559

KTINGSGSPHSKILRQQT
1297
2.557

KTINGRQSPHVKAQNQQT
1298
2.557

KTINGSGSPHSKAQSIKT
1299
2.555

KTINGSGSPHSKAQASKT
1300
2.546

KTINGSGSPHSRLFNQQT
1301
2.542

KTINGSGSPHIYLQNQQT
1302
2.542

KTINGSGSPHSRVRNQQT
1303
2.540

KTINGSGSPHSKAVRAQT
1304
2.538

KTINGSGSPHSKPARQQT
1305
2.538

KTINGSGSPHSRYSNQQT
1306
2.536

KTINGSRSPHRSAQNQQT
1307
2.536

KTINGSLSPHIYAQNQQT
1308
2.536

KTINGSGSPHSKPVRQQT
1309
2.529

KTINGMRSPHGKAQNQQT
1310
2.527

KTINGSGSPHSKARITQT
1311
2.525

KTINGSGSPHSWSLNQQT
1312
2.523

KTINTSRSPHSKAQNQQT
1313
2.520

KTINGSGSPHSKAFTRQT
1314
2.518

KTINGSGSPHSKAVRNQT
1315
2.516

KTINGSGSPHSKAQTNRT
1316
2.510

KTINGSGSPHSKANRMQT
1317
2.508

KTINGSGSPHSKAQLVLT
1318
2.508

KTINGSGSPHSKATRQQT
1319
2.505

KTINGSGSPHSKARGTQT
1320
2.505

KTINGSGSPHSKAQWSVT
1321
2.501

KTINGSGSPHSKAWLIQT
1322
2.499

KTINGSGSPHSKAFRPQT
1323
2.499

KTINGSGSPHRRSQNQQT
1324
2.497

KTINGSGSPHSKGIRQQT
1325
2.497

KTINGSGSPHCTLQNQQT
1326
2.497

KPLPGSGSPHSKAQNQQT
1327
2.495

KTINGSGSPHLVCQNQQT
1328
2.493

KTINGSGSPHSKARGYQT
1329
2.492

KTINGRISPHGKAQNQQT
1330
2.492

KTINGSSSPHWLAQNQQT
1331
2.490

KTINGSGSPHSKARMAQT
1332
2.488

KPLDGSGSPHSKAQNQQT
1333
2.488

KPLRGSGSPHSKAQNQQT
1334
2.486

KTINGSGSPHSKAQNAKL
1335
2.486

KTINGSGSPHSKLSKQQT
1336
2.482

KTINGSGSPHSKARNGQT
1337
2.482

KTINGSGSPHSKAQRRQT
1338
2.479

KTINGSGSPHSKWPGQQT
1339
2.477

KTINGSGSPHAFLQNQQT
1340
2.475

KTINGILSPHRKAQNQQT
1341
2.475

KTINGSGSPHSWGSNQQT
1342
2.473

KTINGSGSPHSSCLNQQT
1343
2.471

KTINGSGSPHSKAQSVKT
1344
2.464

KTINGSGSPHSLRYNQQT
1345
2.462

KTINGSGSPHSKARKLQT
1346
2.458

KHRSGSGSPHSKAQNQQT
1347
2.456

KTINGSGSPHSKWSLQQT
1348
2.453

KTINGSGSPHSKAQTMRT
1349
2.453

KTINGSGSPHSKTIRQQT
1350
2.453

KTINGKLSPHMKAQNQQT
1351
2.449

KTINGSGSPHSKARPFQT
1352
2.443

KTINGSGSPHSKPRVQQT
1353
2.441

KTINGSGSPHSKAQVVLT
1354
2.438

KTRRSSGSPHSKAQNQQT
1355
2.438

KTINGSGSPHSKPSRQQT
1356
2.438

KTINGSGSPHSVYRNQQT
1357
2.432

KTINGSGSPHSKTCSQQT
1358
2.430

KTINGSPSPHRKAQNQQT
1359
2.430

KTINGRSSPHFKAQNQQT
1360
2.428

KTINGSGSPHSKAQMVRT
1361
2.426

KTINGSRSPHCSAQNQQT
1362
2.426

KTINGSCSPHLRAQNQQT
1363
2.423

KTINGSGSPHSKAQCLFT
1364
2.421

KTINGRPSPHVKAQNQQT
1365
2.419

KTINGSGSPHSMFPNQQT
1366
2.419

KTINGMKSPHSKAQNQQT
1367
2.413

KTINGSGSPHSKVRAQQT
1368
2.412

KGLVGSGSPHSKAQNQQT
1369
2.412

KTINGSRSPHVRAQNQQT
1370
2.412

KTIRLRGSPHSKAQNQQT
1371
2.408

KPRLGSGSPHSKAQNQQT
1372
2.406

KTINGSGSPHSKAWYPQT
1373
2.406

KTINGSCSPHVRAQNQQT
1374
2.406

KTINGSGSPHSKRNVQQT
1375
2.404

KTINGSGSPHSKRGLQQT
1376
2.402

KTINGSGSPHSKRLAQQT
1377
2.397

KTINGSGSPHSKVTRQQT
1378
2.397

KTINGSGSPHTWLQNQQT
1379
2.397

KTINGSGSPHSKQRSQQT
1380
2.395

KTINGSGSPHSKAQRCST
1381
2.393

KTINGSPSPHYLAQNQQT
1382
2.391

KTINGRRSPHLKAQNQQT
1383
2.391

KTINGSGSPHWSLQNQQT
1384
2.389

KTINGVTSPHWKAQNQQT
1385
2.387

KTINGSGSPHSKAQTTRT
1386
2.385

KTINGSGSPHSKAQLFKT
1387
2.385

KTINGSGSPHSKARSYQT
1388
2.382

KTINGSGSPHSKLSRQQT
1389
2.380

KTINGSGSPHLVFQNQQT
1390
2.376

KTINGSGSPHSKAQLVKT
1391
2.376

KTINGSGSPHSKAQTGRT
1392
2.371

KTINGSGSPHSKARYSQT
1393
2.369

KTINGRHSPHLKAQNQQT
1394
2.369

KTINGSGSPHSFARNQQT
1395
2.367

KTINGSGSPHSSCQNQQT
1396
2.365

KTINGSGSPHSLFANQQT
1397
2.365

KTINGSGSPHSKRLTQQT
1398
2.363

KTINGSGSPHSKAQTART
1399
2.363

KTINGSGSPHFLNQNQQT
1400
2.359

KTINGSGSPHSKAQLILT
1401
2.358

KEVGGSGSPHSKAQNQQT
1402
2.358

KTINGSRSPHIRAQNQQT
1403
2.356

KTINGSGSPHSSWLNQQT
1404
2.354

KTINGSMSPHLYAQNQQT
1405
2.354

KTINGMSSPHRKAQNQQT
1406
2.352

KTINGSGSPHSKPRPQQT
1407
2.352

KTINGSGSPHSGLWNQQT
1408
2.350

KTINGSGSPHRWAQNQQT
1409
2.350

KTINGSGSPHSKIRLQQT
1410
2.348

KTINGSGSPHSKFSCQQT
1411
2.348

KTINGSGSPHSKSCAQQT
1412
2.348

KTINGSGSPHSKRLMQQT
1413
2.345

KTSRCSGSPHSKAQNQQT
1414
2.345

KTINGSGSPHSFLLNQQT
1415
2.341

KTINGSGSPHCSAQNQQT
1416
2.339

KTINGSGSPHSKAQPSKT
1417
2.339

KTINGSGSPHSYVRNQQT
1418
2.339

KTINGSGSPHSKAQQSRT
1419
2.337

KTINGSGSPHSFVVNQQT
1420
2.335

KTINGFRSPHSKAQNQQT
1421
2.333

KTINGSGSPHSKWLVQQT
1422
2.333

KTINGSGSPHSKRTAQQT
1423
2.331

KTINGLFSPHRKAQNQQT
1424
2.331

KTINTIESPHSKAQNQQT
1425
2.331

KRLFGSGSPHSKAQNQQT
1426
2.330

KTINGSGSPHSKAPNHLT
1427
2.324

KTINGSGSPHLFRQNQQT
1428
2.324

KTINGSGSPHSKASRHQT
1429
2.324

KTINGSGSPHSKLSWQQT
1430
2.322

KETAGSGSPHSKAQNQQT
1431
2.320

KTINGHRSPHLKAQNQQT
1432
2.320

KTINGSGSPHSKGCLQQT
1433
2.320

KTINGSGSPHSKAQVLIT
1434
2.317

KTINGSGSPHSKLRSQQT
1435
2.309

KTINGTLSPHRKAQNQQT
1436
2.307

KTINGSGSPHSTWTNQQT
1437
2.307

KTINGSGSPHSKAQCRLT
1438
2.304

KTINNLRSPHSKAQNQQT
1439
2.302

KTINGSGSPHSKARANQT
1440
2.300

KTINGRQSPHTKAQNQQT
1441
2.294

KTINSARSPHSKAQNQQT
1442
2.292

KTINGCSSPHRKAQNQQT
1443
2.291

KTINGSVSPHFMAQNQQT
1444
2.287

KTINGSGSPHSLCQNQQT
1445
2.285

KTINGSLSPHLFAQNQQT
1446
2.285

KTINGSGSPHSKACPLQT
1447
2.285

KTINGRTSPHRKAQNQQT
1448
2.285

KTINGSGSPHSKRATQQT
1449
2.283

KTINGSGSPHSKARIMQT
1450
2.283

KTINGSGSPHVTWQNQQT
1451
2.281

KTINGSGSPHSKRLPQQT
1452
2.279

KTINGSGSPHSKAQGERT
1453
2.279

KTINGSGSPHLYGQNQQT
1454
2.277

KTINGSGSPHLSCQNQQT
1455
2.277

KTINGSGSPHSKAQFTLT
1456
2.277

KTINGSRSPHFKAQNQQT
1457
2.277

KTINGRPSPHSKAQNQQT
1458
2.276

KTINGFSSPHRKAQNQQT
1459
2.276

KTINGRASPHVKAQNQQT
1460
2.272

KTINGSGSPHSKAQNEVH
1461
2.272

KTINGSGSPHSKRSLQQT
1462
2.270

KTINGSGSPHRSRQNQQT
1463
2.270

KPPTGSGSPHSKAQNQQT
1464
2.270

KTINGSGSPHSKAARTQT
1465
2.266

KTINGSGSPHSSWANQQT
1466
2.264

KTINGSGSPHSKAQRHAT
1467
2.264

KTINSPRSPHSKAQNQQT
1468
2.264

KTINGSGSPHSKSERQQT
1469
2.263

KTINGSGSPHALFQNQQT
1470
2.261

KTINGSGSPHSKAQCYVT
1471
2.261

KTINGVASPHRKAQNQQT
1472
2.261

KTINGSGSPHSALWNQQT
1473
2.261

KTINGSGSPHSKSVRQQT
1474
2.259

KTINGSGSPHSHMENQQT
1475
2.259

KTINGSGTPHSKAQNQQT
1476
2.259

KTINGSGSPHSKTGRQQT
1477
2.259

KTINGSGSPHSKAQANRT
1478
2.255

KTINGSGSPHSKARFSQT
1479
2.253

KYLLGSGSPHSKAQNQQT
1480
2.253

KTINGSGSPHCSSQNQQT
1481
2.253

KTINGSMSPHRKAQNQQT
1482
2.251

KTINGNLSPHRKAQNQQT
1483
2.250

KEVAGSGSPHSKAQNQQT
1484
2.250

KTINLSRSPHSKAQNQQT
1485
2.246

KTINGSGSPHSKARQQQT
1486
2.244

KTINGTPSPHRKAQNQQT
1487
2.244

KTINGSGSPHSKFKLQQT
1488
2.244

KTINGSGSPHSKAWLLQT
1489
2.240

KTINGLRSPHSKAQNQQT
1490
2.238

KTINGRLSPHRKAQNQQT
1491
2.238

KTINGSPSPHLFAQNQQT
1492
2.238

KDLRGSGSPHSKAQNQQT
1493
2.238

KTINGSGSPHSKAQLAKT
1494
2.238

KTINGSGSPHSKPRSQQT
1495
2.235

KTINGSGSPHSKKMSQQT
1496
2.235

KTINGSGSPHSKAQLIVT
1497
2.235

KTINGSGSPHSKARFTQT
1498
2.233

KTINGSGSPHPLFQNQQT
1499
2.233

KTINGSGSPHSKAQRGMT
1500
2.231

KTINGSGSPHSKAQNLRR
1501
2.231

KTINGSGSPHSKAQFRVT
1502
2.231

KTINGSGSPHSKAFVRQT
1503
2.225

KTINGSGSPHSKARLTQT
1504
2.223

KTINGSGSPHRFKQNQQT
1505
2.223

KTINGSGSPHSKEETQQT
1506
2.223

KTINGSGSPHSKTRAQQT
1507
2.223

KTINGSGSPHSVSWNQQT
1508
2.223

KTINGSGSPHTKWQNQQT
1509
2.222

KTINGSNSPHRKAQNQQT
1510
2.218

KTINGSGSPHSKAQNKRS
1511
2.214

KTINGSGSPHSTRQNQQT
1512
2.212

KTINGTRSPHTKAQNQQT
1513
2.203

KTINGSGSPHVLFQNQQT
1514
2.203

KTINGSVSPHYLAQNQQT
1515
2.203

KTINGALSPHRKAQNQQT
1516
2.203

KTINGSGSPHSKARLYQT
1517
2.201

KTINGSGSPHEHNQNQQT
1518
2.199

KTINGVLSPHWKAQNQQT
1519
2.199

KTINGSGSPHSKASRQQT
1520
2.197

KTINGSGSPHSKRSFQQT
1521
2.197

KTINGSGSPHSKRVSQQT
1522
2.196

KTINGSGSPHSYSRNQQT
1523
2.196

KTINGSGSPHSTVWNQQT
1524
2.196

KTINGSGSPHSVLFNQQT
1525
2.194

KTINGPLSPHCKAQNQQT
1526
2.194

KTINGSGSPHSKRVGQQT
1527
2.190

KTINGSGSPHSKLWSQQT
1528
2.190

KTINGSGSPHSKAQGVRT
1529
2.188

KTINGSVSPHRRAQNQQT
1530
2.186

KTINGSGSPHLRFQNQQT
1531
2.186

KTINGSASPHVFAQNQQT
1532
2.186

KTWVRSGSPHSKAQNQQT
1533
2.186

KTINGSGSPHSKARMQQT
1534
2.184

KTINGSGSPHSKASRGQT
1535
2.182

KTINGSGSPHSKAQVCLT
1536
2.182

KTINGSGSPHSKARGVQT
1537
2.181

KTINGSGSPHGLWQNQQT
1538
2.181

KTINGSGSPHSKAQVWFT
1539
2.181

KTINGSGSPHSKAQVTLT
1540
2.179

KTINGSGSPHSKAQLRIT
1541
2.179

KDSLGSGSPHSKAQNQQT
1542
2.175

KTINGSGSPHSKRASQQT
1543
2.173

KTINGSGSPHSKRINQQT
1544
2.173

KTINGSGSPHSKASKNQT
1545
2.171

KTINGSGSPHSKAQLPWT
1546
2.169

KTINGSGSPHSKLTRQQT
1547
2.169

KTINGSGSPHSKINRQQT
1548
2.169

KTINRVISPHSKAQNQQT
1549
2.169

KTINGSGSPHTLWQNQQT
1550
2.168

KTINGSGSPHSRRQNQQT
1551
2.166

KTINGSGSPHSKGGRQQT
1552
2.164

KTINGSESPHDSAQNQQT
1553
2.164

KTINGSGSPHSRPRNQQT
1554
2.164

KTINGSGSPHSRKQNQQT
1555
2.162

KTINGSGSPHSKAQEELT
1556
2.162

KTINGWRSPHSKAQNQQT
1557
2.160

KTINGSGSPHSLLYNQQT
1558
2.158

KTINGSGSPHSFRLNQQT
1559
2.158

KTINGSGSPHSKAQFLRT
1560
2.156

KTINGSGSPHSKQSRQQT
1561
2.156

KTINGSRSPHSKAQNRQT
1562
2.155

KTINGRPSPHIKAQNQQT
1563
2.155

KTINGSGSPHSKRLVQQT
1564
2.151

KGHEGSGSPHSKAQNQQT
1565
2.151

KTINGSGSPHSKAQKRST
1566
2.151

KTINGSGSPHSYLLNQQT
1567
2.147

KTINGSGSPHSKPRGQQT
1568
2.147

KTINGSGSPHSKTRLQQT
1569
2.145

KTINGSGSPHSKSHRQQT
1570
2.145

KEIKGSGSPHSKAQNQQT
1571
2.140

KTINGSGSPHSKARGIQT
1572
2.140

KTINGYRSPHSKAQNQQT
1573
2.140

KTINGSGSPHSKLWTQQT
1574
2.140

KTINGSGSPHSKPWLQQT
1575
2.138

KTINGSGSPHWSVQNQQT
1576
2.138

KTINGSGSPHSKVARQQT
1577
2.136

KTINGSGSPHTLFQNQQT
1578
2.136

KTINGSCSPHLAAQNQQT
1579
2.134

KTINGSGSPHSKTSRQQT
1580
2.132

KTINGSGSPHSKAQNARH
1581
2.127

KTINGSGSPHSKAQLKLT
1582
2.125

KTINGSGSPHSKAQNWRT
1583
2.125

KTINGSGSPHELPQNQQT
1584
2.123

KTINGSGSPHSKNVRQQT
1585
2.123

KTINGSGSPHFMRQNQQT
1586
2.123

KTINGSGSPHGWAQNQQT
1587
2.121

KTINGSGSPHFHLQNQQT
1588
2.121

KTINGSASPHWSAQNQQT
1589
2.121

KTINGSSSPHSWAQNQQT
1590
2.119

KTINGSGSPHSKAHRQQT
1591
2.117

KTINGSGSPHSKQRVQQT
1592
2.117

KTLRRSGSPHSKAQNQQT
1593
2.117

KTINGSGSPHSKGVRQQT
1594
2.115

KTINGSLSPHTWAQNQQT
1595
2.115

KTINGSGSPHSKRALQQT
1596
2.114

KTINGSGSPHCLSQNQQT
1597
2.114

KTINGSGSPHSKAQSLKT
1598
2.110

KTINGSGSPHSFVRNQQT
1599
2.110

KTINGSGSPHSIFSNQQT
1600
2.110

KTINGSGSPHSKVSRQQT
1601
2.108

KTINGSGSPHSKARNKQT
1602
2.108

KTINASGSPHSKAQGQQT
1603
2.108

KTINGSGSPHSKLRMQQT
1604
2.106

KTINGSWSPHMLAQNQQT
1605
2.106

KTINGSGSPHSLFPNQQT
1606
2.106

KPPLGSGSPHSKAQNQQT
1607
2.102

KTINGIASPHRKAQNQQT
1608
2.099

KTINGSCSPHSLAQNQQT
1609
2.099

KTINGRLSPHFKAQNQQT
1610
2.097

KTINGSGSPHSKARMTQT
1611
2.091

KTINGSGSPHSKARLQQT
1612
2.089

KTINGSGSPHSKWVSQQT
1613
2.089

KTINGSGSPHSKKVSQQT
1614
2.088

KTINGSGSPHSKAQSYRT
1615
2.088

KAFNGSGSPHSKAPNQQT
1616
2.088

KTINGSGSPHSKAQYRLT
1617
2.088

KTINGSWSPHLVAQNQQT
1618
2.084

KTINGSGSPHSWTQNQQT
1619
2.084

KTINGSGSPHSKAQSHRT
1620
2.084

KGINGSGSPHGKAQNQQT
1621
2.084

KTINGSGSPHSKAQNRKL
1622
2.084

KTINGRYSPHSKAQNQQT
1623
2.080

KTINGSGSPHSKGRSQQT
1624
2.080

KTINGSGSPHCVAQNQQT
1625
2.080

KTINGSGSPHSKIRPQQT
1626
2.080

KTINGSGSPHSKAQSSKT
1627
2.078

KTINGSGSPHSKRPFQQT
1628
2.076

KTINGSSSPHCLAQNQQT
1629
2.074

KTINGTRSPHAKAQNQQT
1630
2.071

KTINGSGSPHLLFQNQQT
1631
2.069

KTINGRRSPHTKAQNQQT
1632
2.069

KTINGSGSPHSKASKQQT
1633
2.069

KTINGSGSPHSKAQLGRT
1634
2.069

KTINGSGSPHSVFLNQQT
1635
2.069

KTINGSGSPHSKSARQQT
1636
2.067

KTINGSGSPHSKLRLQQT
1637
2.065

KTRKSSGSPHSKAQNQQT
1638
2.065

KTINGFRSPHLKAQNQQT
1639
2.063

KTINGSGSPHSKRSIQQT
1640
2.063

KTINGSGSPHSKGRIQQT
1641
2.061

KTINGSRSPHRPAQNQQT
1642
2.061

KTINGSGSPHSKLRPQQT
1643
2.060

KTINGSGSPHMYAQNQQT
1644
2.060

KTINGRTSPHAKAQNQQT
1645
2.060

KTINGSGSPHSKAGRGQT
1646
2.058

KTINGSGSPHSKLMRQQT
1647
2.056

KTINGSGSPHSKANKSQT
1648
2.056

KTINGSGSPHSKAVRQQT
1649
2.052

KTINGSGSPHSKCLSQQT
1650
2.052

KTINGSGSPHSKAQWVLT
1651
2.052

KTINGSGSPHSKAQFWVT
1652
2.050

KTINGSGSPHSKALCRQT
1653
2.048

KEVMGSGSPHSKAQNQQT
1654
2.047

KTINGSGSPHSKNTRQQT
1655
2.047

KTINGSGSPHTWTQNQQT
1656
2.045

KTINGSTSPHWSAQNQQT
1657
2.043

KTINGNVSPHRKAQNQQT
1658
2.043

KTINGSTSPHLFAQNQQT
1659
2.041

KTINGSGSPHSKAQNYRA
1660
2.039

KTINGSGSPHSKARGQQT
1661
2.039

KTINGSGSPHSKAQNRIR
1662
2.039

KTINGSGSPHSKWTLQQT
1663
2.039

KTINGSGSPHSKAQMKCT
1664
2.039

KTINGSGSPHSLWQNQQT
1665
2.037

KTINGSGSPHSKAQLSKT
1666
2.035

KTINLIWSPHSKAQNQQT
1667
2.035

KTINGSGSPHSKRVLQQT
1668
2.035

KTINGSGSPHSKVRVQQT
1669
2.034

KTINSRFSPHSKAQNQQT
1670
2.032

KRSKGSGSPHSKAQNQQT
1671
2.030

KTINGSGSPHRRLQNQQT
1672
2.030

KTINGSGSPHSCAQNQQT
1673
2.030

KTINGPLSPHRKAQNQQT
1674
2.028

KTINGSVSPHLYAQNQQT
1675
2.028

KTINGRISPHLKAQNQQT
1676
2.028

KTINGSHSPHRKAQNQQT
1677
2.028

KTINGSGSPHSKAQVSIT
1678
2.028

KTINGSMSPHRRAQNQQT
1679
2.028

KTINGRQSPHAKAQNQQT
1680
2.026

KTINGSGSPHSKAVWRQT
1681
2.026

KQPLGSGSPHSKAQNQQT
1682
2.024

KTINGSGSPHSKAQNVKL
1683
2.024

KTINGSGSPHSKRGTQQT
1684
2.022

KTINGSVSPHYVAQNQQT
1685
2.022

KTINGSGSPHSKNLRQQT
1686
2.022

KTINGSGSPHSKAQAFRT
1687
2.022

KTINGSGSPHSKCSNQQT
1688
2.020

KELVGSGSPHSKAQNQQT
1689
2.019

KTINGSGSPHSLVFNQQT
1690
2.019

KTINGSGSPHSKAQATRT
1691
2.019

KTINGTSSPHCKAQNQQT
1692
2.017

KTINGSGSPHSKALWRQT
1693
2.015

KTINGSGSPHSKAQFSVT
1694
2.015

KTINGSGSPHSKLYMQQT
1695
2.015

KTINGSLSPHYMAQNQQT
1696
2.015

KTINGSGSPHSKAWLMQT
1697
2.015

KTINGSGSPHSKSLKQQT
1698
2.013

KTINGSGSPHSKAQNTRR
1699
2.013

KTINGSGSPHYLLQNQQT
1700
2.011

KTINGSGSPHTWSQNQQT
1701
2.011

KTINGSGSPHSKTRMQQT
1702
2.011

KTINTRPSPHSKAQNQQT
1703
2.011

KTINGSGSPHSKAQILVT
1704
2.009

KTINGSGSPHSKAQNAKS
1705
2.009

KTINGSGSPHSRTYNQQT
1706
2.009

KTINGSGSPHSKKGGQQT
1707
2.009

KTINGYSSPHRKAQNQQT
1708
2.007

KTINGSGSPHWVSQNQQT
1709
2.007

KTINGSGSPHSKARLAQT
1710
2.007

KTINGMCSPHSKAQNQQT
1711
2.006

KTINGSGSPHSKSNKQQT
1712
2.006

KTINGSGSPHSKAQFVLT
1713
2.006

KTINGSISPHFVAQNQQT
1714
2.006

KTINGSGSPHRRMQNQQT
1715
2.004

KTINGSGSPHSKAWILQT
1716
2.004

KTINGSGSPHSKAQGVKT
1717
2.002

KTINGSGSPHSKAQFSLT
1718
2.000

KTINMLRSPHSKAQNQQT
1719
2.000

KTINGSGSPHSKAQLGKT
1720
2.000

KTINGSGSPHSMYLNQQT
1721
2.000

As shown in Table 14, approximately 72 TTM-002 matured capsid variants demonstrated at least a 2-fold increase in expression relative to the non-matured TTM-002 control, with a few variants demonstrating greater than a three- to five-fold enrichment relative to the non-matured TTM-002 control. Also, across the peptides comprised within the TTM-002 matured capsid variants with the greatest fold-enrichment relative to the non-matured TTM-002 capsid in the brain, it was observed that the modifications in the variant sequences appeared in the region N-terminal to the SPH motif present within the capsid variant. This indicates that modifications that appeared to improve TTM-002 capsid tropism in the CNS of mice were skewed to the N-terminal portion of the peptide insertion in loop IV of the sequence. Additionally, a number of these N-terminal modifications that were incorporated into the matured TTM-002 capsid variants were negatively charged amino acids (particularly glutamic acid (E)).

TABLE 14

NGS fold-enrichment of TTM-002 matured AAV

capsid variants in the brain of

CD-1 Outbred mice

Fold

SEQ
enrichment

ID
over

Peptide Sequence
NO:
TTM-002

KTINGHDSPHVTDQNQQT
1722
5.20

KTINGHDSPHKRGQHRQT
1723
4.20

KMPEGHDSPHKSGQNQQT
1724
3.18

KMEGGHDSPHKSGQNQQT
1725
2.72

KMEYGHDSPHKSGQNQQT
1726
2.71

KAEWGHDSPHKSGQNQQT
1727
2.69

KCEWGHDSPHKSGQNQQT
1728
2.68

KANNGQDSPHKSGQNQQT
1729
2.67

KTINGHDSPHLCGQNQQT
1730
2.59

KIPEGHDSPHKSGQNQQT
1731
2.54

KADMGHDSPHKSGQNQQT
1732
2.48

KTINGHLSPHYFGQNQQT
1733
2.41

KIEYGHDSPHKSGQNQQT
1734
2.41

KADYGHDSPHKSGQNQQT
1735
2.40

KIETGHDSPHKSGQNQQT
1736
2.38

KTINGHDSPHTNGQKQQT
1737
2.38

KMEWGHDSPHKSGQNQQT
1738
2.38

KTINGHDSPHWLLQNQQT
1739
2.37

KCEYGHDSPHKSGQNQQT
1740
2.36

KRINGHDSPHKSGQKQQN
1741
2.34

KMEIGHDSPHKSGQNQQT
1742
2.34

KLEYGHDSPHKSGQNQQT
1743
2.33

KADWGHDSPHKSGQNQQT
1744
2.32

KIEIGHDSPHKSGQNQQT
1745
2.30

KTIKDNDSPHKSGQNQQT
1746
2.27

KDIMGHDSPHKSGQNQQT
1747
2.23

KFEQGHDSPHKSGQNQQT
1748
2.22

KMEFGHDSPHKSGQNQQT
1749
2.21

KCDQGHDSPHKSGQNQQT
1750
2.21

KLPEGHDSPHKSGQNQQT
1751
2.19

KIENGHDSPHKSGQNQQT
1752
2.19

KMESGHDSPHKSGQNQQT
1753
2.18

KAEIGHDSPHKSGQNQQT
1754
2.17

KVEYGHDSPHKSGQNQQT
1755
2.17

KIINGHDSPHKSGLTQQT
1756
2.17

KTSNGDDSPHKSGRNQQT
1757
2.17

KIEVGHDSPHKSGQNQQT
1758
2.16

KMEMGHDSPHKSGQNQQT
1759
2.16

KAEVGHDSPHKSGQNQQT
1760
2.15

KMDAGHDSPHKSGQNQQT
1761
2.15

KVEWGHDSPHKSGQNQQT
1762
2.15

KAEQGHDSPHKSGQNQQT
1763
2.14

KLEWGHDSPHKSGQNQQT
1764
2.14

KTINGHPSPHYLGQNQQT
1765
2.14

KTINGHLSPHYYGQNQQT
1766
2.13

KMELGHDSPHKSGQNQQT
1767
2.13

KMETGHDSPHKSGQNQQT
1768
2.12

KMEAGHDSPHKSGQNQQT
1769
2.12

KTINGHDSPHLLWQNQQT
1770
2.12

KTINRQRSPHKSGQNQQT
1771
2.11

KIESGHDSPHKSGQNQQT
1772
2.11

KTAKDHDSPHKSGQNQQT
1773
2.11

KMEVGHDSPHKSGQNQQT
1774
2.11

KCEIGHDSPHKSGQNQQT
1775
2.10

KATNGHDSPHKSGLNQQT
1776
2.10

KMDGGHDSPHKSGQNQQT
1777
2.09

KQEVGHDSPHKSGQNQQT
1778
2.07

KADQGHDSPHKSGQNQQT
1779
2.07

KTINGHESPHKSAQNHQT
1780
2.07

KTINGHDSPHKSAQNQWT
1781
2.07

KNMNGHDSPHKSGQNTHS
1782
2.06

KTPWEHDSPHKSGQNQQT
1783
2.05

KTINGHSSPHYFGQNQQT
1784
2.05

KIEMGHDSPHKSGQNQQT
1785
2.05

KTANEHDSPHKSGQNQQT
1786
2.05

KTINGHDSPHKSGRRRQT
1787
2.04

KISNGHDSPHKSAQNQQT
1788
2.03

KTGNGHDSPHKSGQYQQT
1789
2.03

KTINGHYSPHLFGQNQQT
1790
2.02

KTINGNYSPHKIGQNQQT
1791
2.02

KTINGHDSPHKSRQNDQT
1792
2.01

KQQQGHDSPHKSGQNQQT
1793
2.01

KTPQDHDSPHKSGQNQQT
1794
2.00

KHDWGHDSPHKSGQNQQT
1795
2.00

KIEGGHDSPHKSGQNQQT
1796
2.00

These data demonstrate that following two maturation approaches, matured TTM-001 and TTM-002 capsid variants with loop IV modifications were generated with significantly enhanced CNS tropism in mice compared to the corresponding non-matured TTM-001 and TTM-002 capsid variants, which already exhibited a significant fold enrichment over AAV9 in the mouse brain.

Example 4. Maturation of TTM-001 and TTM-002 Capsid in NHPs

This Example describes maturation of the AAV9 capsid variants, TTM-001 (SEQ ID NO: 981 (amino acid) and 983 (DNA), comprising SEQ ID NO: 941 (encoded by SEQ ID NO: 942)) and TTM-002 (SEQ ID NO: 982 (amino acid) and 984 (DNA), comprising SEQ ID NO: 2 (encoded by SEQ ID NO: 3)) in NHPs to further enhance their transduction and biodistribution in the central nervous system as well as other tissues, and evolve the AAV capsid variants to provide further cross-species compatibility. Two approaches were used to mature the TTM-001 and TTM-002 capsid sequences in order to randomize and mutate within and around the peptide insert comprised within loop IV of the capsid variant. As many of the AAV capsid variants that demonstrated the greatest fold-enrichment in the NHP brain relative wild-type AAV9 comprised an SPH motif in the same position (e.g., immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138) (see Example 1), the SPH motif was not mutated in either approach to mature the TTM-001 and TTM-002 capsid variants. In the first maturation approach, sets of three contiguous amino acids were randomized across the mutagenesis region in the TTM-001 and TTM-002 sequences, which spanned from position 450 to position 466, numbered according to SEQ ID NO: 981 and 982. In the second maturation approach, mutagenic primers were used to introduce point mutations at a low frequency, scattered across the mutagenesis region in the TTM-001 and TTM-002 sequences ranging from position 449 to position 466, numbered according to SEQ ID NO: 981 and 982. AAV capsid variants arising from each maturation approach for TTM-001 and TTM-002 were pooled together, for subsequent testing and characterization in NHPs.

The library of pooled matured AAV capsid variants generated using the first maturation approach and the second maturation approach for the TTM-001 and TTM-002 AAV capsid variants were injected into two NHPs. After a period in life, the brains, heart, liver, muscle, and DRG of the NHPs were isolated and RNA was extracted. Following RNA recovery and RT-PCR amplification, a systematic NGS enrichment analysis was performed to calculate the fold enrichment ratio relative to an AAV9 control, and the peptides comprised within the variants were identified.

Following the RNA recovery and NGS analysis from the second maturation approach, approximately 680,000 capsid variants were identified. The 680,000 matured capsid variants were then filtered based on samples with a raw virus count greater than 10 and a coefficient of variance (CV) of less than 1, which was calculated for each peptide across the brain samples taken from the two NHPs. Those that had a CV value <1 were identified, as these were the peptides that were reliably detected in the majority of samples isolated from the brains of the two NHPs. Using this filtering criteria, this led to approximately 64,000 matured capsid variants.

Table 15 provides the peptide sequences of the matured capsid variants having a raw virus count greater than 10, a CV of less than 1 for the brain samples isolated, and that also demonstrated a 50-fold or greater fold-increase in expression in the brain relative to the AAV9 control in both mice and NHPs. The matured variants in Table 15, were also those variants that had a fold-change in expression that was less than 2 relative to the AAV9 control in the liver and the DRG. Applying these criteria, approximately 350 matured capsid variants were identified that demonstrated high transduction in the brain in NHPs and mice, cross-species compatibility in mice and NHPs, and were de-targeted in the liver and DRG, relative to the AAV9 control. Several variants as shown in Table 15, led to greater than 100-fold increase in expression relative to AAV9 in the NHP and/or mouse brain, with one variant resulting in a greater than 200-fold increase in expression relative to AAV9 in both species.

Fold-change in expression for the TTM-001 and TTM-002 matured variants in Table 15 that showed increased expression in the brain of the NHPs and mice, were also calculated for the DRG, muscle, liver (RNA and DNA), and heart of the NHPs following each maturation approach. As shown in Table 15, many variants were de-targeted in the peripheral tissues with a lower fold-change in expression relative to the AAV9 control, demonstrating CNS-specific tropism and a preferential transduction of the brain and CNS. Some variants demonstrated increased expression to AAV9 in multiple tissues, including the brain and peripheral tissues, demonstrating pan-tropism.

TABLE 15

NGS fold-enrichment of TTM-001 and TTM-002 matured AAV capsid variants

in th ebrain of NHPs and mice

Fold Enrichment relative to AAV9

SEQ

Liver
Liver

ID
Brain
DRG
Heart
Muscle
RNA
DNA
Brain

Sequence
NO:
(NHP)
(NHP)
(NHP)
(NHP)
(NHP)
(NHP)
(Mouse)

KTIIGSGSPHSKAQNRHT
3239
217.176
0.000
0.000
0.000
0.000
0.000
210.515

KTFPGSGSPHSKVQNQQT
3240
199.720
0.000
0.000
0.000
0.000
0.967
97.703

KTEKMSGSPHSKAQNQQT
3241
169.461
0.523
0.000
0.000
0.000
0.158
109.161

KEINGRGSPHSKAQNQQT
3527
134.390
0.239
0.000
0.000
0.000
0.232
52.311

KTVNRNGSPHSKAQNQQT
3528
133.016
0.000
0.416
0.000
0.000
0.000
85.361

KTVNGSGSPHSKARDQQT
3242
124.789
0.123
0.039
0.312
0.569
0.454
132.137

KTFNGSGSPHSKAPNLQT
3243
121.436
0.000
0.167
0.000
0.000
0.015
168.920

KTEKTSGSPHSKAQNQQT
3244
120.337
0.000
0.355
0.000
0.000
0.119
101.467

KTINGSGSPHSKAHVRQT
3245
119.798
0.000
0.000
0.262
0.694
1.039
165.590

KTVNGSGSPHSKAPNQHT
3246
117.207
0.000
0.109
0.000
0.000
0.074
51.008

KTEKISGSPHSKAQNQQT
3247
116.603
0.000
0.000
0.000
0.000
0.426
102.978

KTINGPGSPHSKAHNQQT
3529
115.742
0.146
0.000
0.235
0.000
0.513
52.508

KTVNGSGSPHSKTQSQQT
3248
115.086
0.000
0.726
0.000
0.000
0.340
63.248

TTINGSGSPHSKAQNQQT
3249
114.856
1.340
14.856
0.827
1.281
0.957
72.058

KSINESGSPHSKAQNQQI
3250
113.833
0.000
0.000
0.000
0.000
0.000
67.649

KTERTSGSPHSKAQNQQT
3251
112.957
0.000
0.009
0.000
1.128
0.207
117.374

KTINGSGSPHSKAQPAKT
3252
111.472
0.331
0.000
1.089
0.044
1.796
215.275

KTEKSSGSPHSKAQNQQT
3253
107.470
0.000
0.016
0.014
0.977
0.179
100.177

KTSYGNGSPHSKAQNQQT
3530
105.937
0.000
0.000
0.000
0.000
0.114
105.894

KTEKGSGSPHSKAQNQQT
3254
105.614
0.053
0.031
0.000
0.586
0.169
84.653

KTINGSGSPHSKSQTQQN
3255
104.474
0.000
0.131
0.000
0.084
0.038
54.021

KTERISGSPHSKAQNQQT
3256
103.692
0.000
0.000
0.000
0.062
0.370
89.637

KTERASGSPHSKAQNQQT
3257
103.669
0.000
0.000
0.000
0.127
0.070
115.550

KELHGSGSPHSKAQNQQT
3258
102.680
0.000
0.000
0.000
1.634
0.592
96.554

KAINGSGSPHSKAQNLAT
3259
101.954
0.000
10.954
8.655
0.298
0.239
116.685

KTVNGSGSPHSKSQNQLT
3260
101.327
0.000
0.035
0.000
0.000
0.025
80.716

KTERNSGSPHSKAQNQQT
3261
99.892
0.000
0.000
0.000
0.000
0.107
87.392

KSVNGNGSPHSKAQNQQT
3531
99.385
0.000
1.329
0.000
0.359
0.079
51.016

KTFNGSGSPHSKAQGQQT
3262
99.253
0.000
0.208
0.000
0.128
0.099
81.459

KTINGSGSPHGWVQNQQT
3532
97.122
0.000
0.000
0.000
1.240
1.975
290.720

KTERVSGSPHSKAQNQQT
3263
96.943
0.000
0.000
0.000
0.000
0.144
135.438

KTINGSGSPHSKALNRQS
3264
96.843
0.136
0.532
0.000
0.042
0.178
55.945

KTERLSGSPHSKAQNQQT
3265
95.857
0.000
0.004
0.005
0.126
0.260
102.372

KTDNGSGSPHSKAHNQQT
3266
95.164
0.000
0.000
0.000
0.000
0.027
55.313

KTFHGSGSPHSKTQNQQT
3267
94.714
0.000
0.210
0.120
0.000
0.000
51.119

KTINGGGSPHSKAQTQQI
3533
92.345
0.000
0.000
0.000
0.000
0.023
54.199

KTSNGSGSPHSKAQNPPT
3268
91.528
0.000
0.000
0.000
0.000
0.039
51.541

ETINGSGSPHSKAQNLQT
3269
90.969
0.221
1.023
0.197
0.179
0.813
107.216

KTVHGNGSPHSKAQNQQT
3534
90.073
0.000
0.000
0.000
0.000
0.304
97.003

NTINGSGSPHSKAQNQQT
3270
90.017
1.712
1.261
1.171
0.923
0.540
55.179

KTINGGGSPHSKAQNQQC
3535
89.301
0.219
0.000
0.000
0.287
0.319
53.840

KTENMSGSPHSKAQNQQT
3271
89.247
0.000
0.000
0.000
0.000
0.260
130.568

KTENVSGSPHSKAQNQQT
3272
88.506
0.000
0.000
0.000
0.964
0.112
108.591

KTSSGSGSPHSKAQYQQT
3273
87.304
0.000
0.000
0.000
0.000
0.299
58.143

KTIDGGGSPHSKAQNKQT
3536
85.019
0.000
0.000
0.000
0.000
0.477
55.517

KTEKVSGSPHSKAQNQQT
3274
84.558
0.000
0.022
0.000
0.873
0.424
112.185

KAINGSGSPHSKAQDQET
3275
84.080
0.000
0.000
0.000
0.194
0.027
87.637

KTCNKSGSPHSKAQNQQT
3276
83.992
0.000
0.000
0.165
0.283
0.000
119.496

KTINGGGSPHSKAQNQLI
3537
83.881
0.000
0.000
0.000
0.046
0.387
78.383

KNINGGGSPHSKAQNQQT
3538
83.083
0.000
0.042
0.000
0.000
0.000
75.913

KTEHLSGSPHSKAQNQQT
3277
83.080
0.000
0.000
0.012
0.021
0.189
69.494

KAEMGSGSPHSKAQNQQT
3278
83.049
0.000
0.020
0.000
0.768
0.112
135.019

KATNGSGSPHSKAQNHQT
3279
82.627
0.000
0.176
0.000
0.155
0.057
66.207

KAIKGSGSPHSKAQDQQT
3280
82.258
0.000
0.000
0.000
0.108
0.000
85.178

KTINGGGSPHSKSQNQLT
3539
82.231
0.000
0.070
0.000
0.000
0.498
126.986

KTVNGNGSPHSKAQNKQT
3540
81.481
0.000
0.000
0.000
0.000
0.122
69.455

KTINGSGSPHSKGHWQQT
3281
81.434
0.000
0.000
0.000
0.000
1.011
65.252

KTDKTSGSPHSKAQNQQT
3282
81.430
0.000
0.000
0.000
1.362
0.291
169.515

KTFKGSGSPHSKAPNQQT
3283
80.890
0.000
0.000
0.000
0.000
0.017
71.144

KTVNGSGSPHSKAQNQLI
3284
80.509
0.000
0.000
0.000
0.000
0.166
71.156

KTINGSGSPHSKRPEQQT
3285
80.418
0.000
0.013
0.000
0.149
0.361
50.319

KTINGSGSPHSKAQRTMT
3286
80.388
0.000
0.022
0.170
1.812
1.025
100.248

KTEKASGSPHSKAQNQQT
3287
80.285
0.000
0.041
0.000
0.000
0.261
90.390

KSDQGSGSPHSKAQNQQT
3288
80.076
0.000
0.000
0.000
0.993
0.124
151.911

KTEITSGSPHSKAQNQQT
3289
79.620
0.000
0.163
0.000
0.332
0.074
76.686

KTDKSSGSPHSKAQNQQT
3290
79.470
0.055
0.012
0.000
1.437
0.367
141.351

KTIDGSGSPHSKAQNQQH
3291
79.090
0.000
0.000
0.000
0.136
0.049
57.914

KTVNGNGSPHSKAQNQHT
3541
78.849
0.000
0.000
0.000
0.000
0.045
54.086

KNINGSGSPHSKAQNQQT
3292
78.445
0.000
0.000
0.000
0.571
0.177
89.719

KTETHSGSPHSKAQNQQT
3293
77.974
0.000
0.067
0.000
0.000
0.512
57.287

KTINGGGSPHSKALNQQN
3542
77.822
0.000
0.131
0.000
0.000
0.274
69.884

KTINGSGSPHSKALHQHT
3294
77.502
0.000
0.052
0.041
0.000
0.188
68.196

KTINGTGSPHSKAQNHQI
3543
77.089
0.171
0.000
0.000
0.000
0.166
54.281

KTINGSGSPHSKAQHRIT
3295
76.849
0.105
0.499
0.170
1.424
0.214
127.000

KTINGSGSPHSKAQYIHT
3296
76.170
0.000
0.014
0.033
1.523
0.168
59.649

KTENISGSPHSKAQNQQT
3297
76.072
0.000
0.000
0.000
0.115
0.132
83.118

KTIIGGGSPHSKAHNQQT
3544
75.872
0.000
0.050
0.000
0.000
0.235
65.492

KTINGSGSPHSKAQKFET
3298
75.788
0.000
0.000
0.028
0.108
0.093
65.588

KTSNESGSPHSKAQNHQT
3299
75.720
0.000
0.000
0.000
0.169
0.217
70.590

KTINGSGSPHSKAQFPST
3300
75.677
0.000
0.004
0.000
0.849
0.127
119.712

KTERPSGSPHSKAQNQQT
3301
75.669
0.000
0.029
0.000
0.000
0.156
73.894

KTINGNGSPHSKAQNPLT
3545
75.269
0.000
0.000
0.000
0.366
0.000
53.583

KSIKGNGSPHSKAQNQQT
3546
75.196
0.000
0.000
0.000
0.000
0.000
90.251

KTERMSGSPHSKAQNQQT
3302
74.910
0.000
0.000
0.000
0.100
0.151
122.812

KTERSSGSPHSKAQNQQT
3303
74.853
0.000
0.071
0.000
1.036
0.056
125.538

KTELHSGSPHSKAQNQQT
3304
74.620
0.000
0.000
0.000
0.021
0.089
53.124

KTELTSGSPHSKAQNQQT
3305
74.548
0.000
0.000
0.000
0.537
0.421
100.311

KTINGSGSPHSKAHNQQR
3306
74.272
0.562
0.486
0.047
0.956
0.057
107.301

KTINGGGSPHSKAQSQQI
3547
74.264
0.000
0.000
0.000
0.000
0.235
67.651

KTINGSGSPHSKAQAIKT
3307
74.261
0.255
0.000
0.000
0.186
0.132
73.560

KTENTSGSPHSKAQNQQT
3308
74.061
0.000
0.000
0.218
0.233
0.730
96.249

KTIDGSGSPHSKGQNRQT
3309
73.930
0.000
0.000
0.000
0.106
0.091
63.626

KNINGSGSPHSKAQSQQT
3310
73.757
0.000
0.000
0.000
0.000
0.041
57.432

KTINGSVSPHGKAQNQLT
3548
73.525
0.000
0.061
0.067
0.000
0.053
51.358

KTSNASGSPHSKAQNQLT
3311
73.501
0.000
0.000
0.297
0.000
0.313
150.401

KTEARSGSPHSKAQNQQT
3312
73.349
0.000
0.000
0.000
0.695
0.118
62.903

KTEKNSGSPHSKAQNQQT
3313
73.347
0.000
0.000
0.044
0.159
0.021
74.393

KTANGSGSPHSKAQYQQT
3314
73.038
0.000
0.000
0.000
0.153
0.160
139.451

KTVNGSGSPHSKAQYQHT
3315
72.847
0.000
0.000
0.000
0.000
0.130
54.158

KTINGSGSPHTKAQNPQS
3316
72.594
0.000
0.000
0.000
0.000
0.130
62.508

KTINGSGSPHSKGQNPPT
3317
72.339
0.000
0.206
0.000
0.000
0.041
134.808

KTIIGSGSPHSKAQHQLT
3318
72.291
0.000
0.000
0.000
0.000
0.000
100.144

KTINGSGSPHSKAQSPPT
3319
71.632
0.069
0.047
0.274
0.179
0.425
97.111

NTIYGSGSPHSKAQNQQT
3320
71.267
1.739
0.000
273.69
0.000
0.209
59.707

KTINGSGSPHSKAQAKLT
3321
71.154
0.000
0.273
0.017
1.591
0.777
130.132

KTDKNSGSPHSKAQNQQT
3322
70.964
0.000
0.000
0.000
0.070
0.123
62.932

KTINGSGSPHSKTKSQQT
3323
70.891
0.000
0.568
0.045
0.418
0.496
83.923

KTINGSGSPHSKAQDRPT
3324
70.831
0.132
0.006
0.000
0.039
0.379
66.800

KTINGIGSPHSKAQNLGT
3549
70.543
0.000
0.071
0.000
0.000
0.135
104.769

KTINGSGSPHSKAQSQQL
3325
70.539
0.000
0.000
0.000
0.000
0.041
51.126

KTENLSGSPHSKAQNQQT
3326
70.303
0.070
0.000
0.000
0.395
0.470
107.385

KTINGSGSPHSKAQAFHT
3327
70.159
0.033
0.000
0.058
0.762
0.119
86.268

KTINGSGSPHSKAQKQQD
3328
70.116
0.000
0.024
0.000
0.064
0.083
110.196

KTFSGSGSPHSKAQNLQT
3329
70.035
0.000
0.327
0.303
0.000
0.228
70.917

KAINGSGSPHSKAQNAQT
3330
69.651
0.000
0.000
0.000
0.023
0.142
72.160

KTESWSGSPHSKAQNQQT
3331
69.144
0.000
0.000
0.000
0.000
0.019
67.699

KTTNGSGSPHSKAHNQLT
3332
69.062
0.000
0.000
0.000
0.708
0.000
65.505

KTVNGNGSPHSKAQNHQT
3550
68.889
0.000
0.000
0.000
0.000
0.030
52.482

KTEDKSGSPHSKAQNQQT
3333
68.813
0.000
0.000
0.000
0.000
0.000
70.071

KTESASGSPHSKAQNQQT
3334
68.651
0.000
0.000
0.000
0.274
0.084
80.500

KTNNGSGSPHSKAQNQQY
3335
68.530
0.000
0.040
0.000
0.000
0.059
82.656

KTSNGGGSPHSKAQNLQT
3551
68.311
0.000
0.052
0.000
0.000
0.000
124.871

KTDKMSGSPHSKAQNQQT
3336
68.167
0.000
0.000
0.000
0.017
0.205
88.234

KEVHGSGSPHSKAQNQQT
3337
67.901
0.000
0.000
0.000
0.727
0.000
100.111

KTINGSGSPHSKAQKLNT
3338
67.782
0.073
0.092
0.000
1.232
0.201
68.637

KTINGGGSPHSKSQNQHT
3552
67.773
0.000
0.057
0.000
0.000
0.220
100.748

KTVNGGGSPHSKAQSQQT
3553
67.634
0.000
0.055
0.000
0.000
0.210
160.711

KTTNGSGSPHSKAQYQHT
3339
67.325
0.000
0.000
0.000
1.378
0.080
83.337

KTISGSGSPHSKAQYQHT
3340
66.739
0.000
0.000
0.000
0.000
0.191
59.822

KTESTSGSPHSKAQNQQT
3341
66.649
0.000
0.009
0.000
1.688
0.176
95.861

KTINGSGSPHSKSQNVQT
3342
66.627
0.000
0.190
0.000
0.202
0.188
56.672

KSINGSGSPHSKAQAQQT
3343
66.464
0.000
0.711
0.000
0.148
0.111
78.451

KTVNGSGSPHSKAQNLQA
3344
66.379
0.000
0.000
0.000
0.000
0.132
50.934

KTVRDSGSPHSKAQNQQT
3345
66.056
0.000
0.025
0.000
0.129
0.461
142.600

KTFNASGSPHSKAPNQQT
3346
65.392
0.208
0.000
0.000
0.215
0.156
66.275

KTDRMSGSPHSKAQNQQT
3347
65.143
0.000
0.000
0.000
0.332
0.103
104.890

KTINGSGSPHSKAQTPPT
3348
64.657
0.010
0.015
0.014
0.200
0.207
54.179

ETIKGSGSPHSKAQNQQT
3349
64.609
0.000
0.000
0.144
0.000
0.024
67.201

KNHIGSGSPHSKAQNQQT
3350
64.535
0.000
0.000
0.000
1.253
0.187
70.356

KTINGSGSPHSKAQYQHA
3351
64.435
0.000
0.000
0.024
0.993
0.097
57.278

KTIPIDGSPHSKAQNQQT
3554
64.421
0.000
0.047
0.000
0.234
0.936
76.826

KTINGSGSPHSKAQGQQA
3352
64.128
0.000
0.185
0.000
0.063
0.195
64.116

KTFNGSGSPHNKAQNHQT
3353
64.060
0.000
0.000
0.035
0.094
0.317
67.757

KESDGSGSPHSKAQNQQT
3354
63.766
0.000
0.000
0.000
0.567
0.146
115.231

KTINGSGSPHSKAQPPAT
3355
63.510
0.048
0.030
0.031
0.126
0.302
117.453

KTINGSGSPHSKAQERPT
3356
63.460
0.000
0.011
0.000
0.810
0.173
57.506

KTIKGSGSPHSKAQDLQT
3357
63.260
0.000
0.000
0.000
0.000
0.218
58.576

KTDLKSGSPHSKAQNQQT
3358
63.152
0.000
0.000
0.012
0.285
0.377
62.687

KTINGGGSPHSKAQNPPT
3555
63.041
0.000
0.082
0.000
0.000
0.057
64.045

KTINGSGSPHSKAQAMHT
3359
62.756
0.000
0.000
0.010
0.976
0.393
84.056

KTVPNSGSPHSKAQNQQT
3360
62.540
0.000
0.000
0.011
0.202
0.161
93.793

KTVIGSGSPHSKALNQQT
3361
62.358
0.000
0.310
0.000
0.062
0.245
60.369

KTINGSGSPHSKAQHPST
3362
62.255
0.000
0.044
0.000
1.345
0.301
101.103

KTINGLGSPHSKSQNQQT
3556
62.170
0.000
0.157
0.000
0.146
0.107
64.139

KTINGTGSPHSKAQNQQM
3557
62.151
0.000
0.000
0.000
0.000
0.000
62.376

KTINGSGSPHSKAPGLQT
3363
62.043
0.007
0.000
0.005
0.651
0.210
144.610

KTINGSGSPHSKAQGIRT
3364
61.952
0.041
0.000
0.012
0.897
0.502
155.013

KTESHSGSPHSKAQNQQT
3365
61.947
0.000
0.000
0.000
1.480
0.106
52.506

KTINGSGSPHSKAQAPAT
3366
61.934
0.000
0.169
0.015
0.696
0.197
127.420

KTINGSGSPHSKSQSQQI
3367
61.870
0.000
0.000
0.000
0.200
0.175
64.027

KAEHGSGSPHSKAQNQQT
3368
61.830
0.000
0.000
0.000
0.772
0.184
116.201

KTEDRSGSPHSKAQNQQT
3369
61.756
0.000
0.000
0.000
1.004
0.408
66.887

KNCLGSGSPHSKAQNQQT
3370
61.442
0.000
0.036
0.000
1.849
0.026
82.488

KTDRGSGSPHSKAQNQQT
3371
61.419
0.000
0.004
0.000
0.211
0.316
74.256

KTINGSGSPHSKAQIPPT
3372
61.258
0.000
0.000
0.000
0.758
0.115
87.661

KTVKGSGSPHSKAQDQQT
3373
61.175
0.000
0.041
0.000
0.432
0.090
58.114

KNADGSGSPHSKAQNQQT
3374
60.944
0.000
0.000
0.000
1.239
0.085
104.503

KTDKVSGSPHSKAQNQQT
3375
60.935
0.000
0.015
0.000
0.765
0.128
146.657

KTITGSGSPHSKAQTQLT
3376
60.846
0.160
8.992
0.000
0.000
0.000
55.640

KTINGSGSPHSKAQAPST
3377
60.696
0.200
0.005
0.000
0.751
0.263
115.528

KNCVGSGSPHSKAQNQQT
3378
60.535
0.000
0.000
0.000
0.018
0.282
96.175

KTIRDAGSPHSKAQNQQT
3558
60.346
0.000
0.000
0.000
0.141
0.251
113.179

KTVKDSGSPHSKAQNQQT
3379
60.216
0.000
0.019
0.000
0.443
0.251
87.334

KNALGSGSPHSKAQNQQT
3380
60.014
0.000
0.003
0.000
0.682
0.213
137.222

KVINGSGSPHSKGQNQQT
3381
60.001
0.000
0.000
0.031
0.264
0.157
68.532

KTVNGGGSPHSKAQNQQS
3559
59.871
0.062
0.020
0.000
0.080
0.185
61.847

KTIQDGGSPHSKAQNQQT
3560
59.865
0.000
0.000
0.116
1.435
0.789
87.522

KTISGGGSPHSKAQNQQN
3561
59.801
0.000
0.000
0.000
0.722
0.039
87.761

KTSNASGSPHSKAHNQQT
3382
59.607
0.000
0.078
0.067
0.031
0.050
67.967

KTINGSGSPHSKAQNTYA
3383
59.603
0.000
0.000
0.000
0.425
0.346
101.715

KTINGSGSPHSKSQNQHI
3384
59.438
0.000
0.099
0.000
0.111
0.108
76.025

KTINGGGSPHSKAQDKQT
3562
59.322
0.000
0.000
0.000
0.000
0.093
50.764

KTEFVSGSPHSKAQNQQT
3385
59.306
0.000
0.000
0.000
0.196
0.276
69.788

KTVNGSGSPHSKAQNHLT
3386
59.239
0.133
0.034
0.000
0.000
0.156
70.786

KTREISGSPHSKAQNQQT
3387
59.027
0.000
0.042
0.224
0.356
0.269
51.696

KTINGSGSPHSKAQIGMT
3388
59.013
0.081
106.528
0.000
1.003
0.248
134.585

KTIDGSGSPHSKALNKQT
3389
58.992
0.000
0.267
0.000
0.000
0.056
74.626

KTIIGGGSPHSKAQNPQT
3563
58.924
0.000
0.202
0.000
0.000
0.126
53.992

KQGEGSGSPHSKAQNQQT
3390
58.752
0.000
0.000
0.000
0.000
0.151
135.300

KTINGTGSPHSKAPNQLT
3564
58.738
0.000
0.000
0.000
0.229
0.035
86.939

KTVNGSGSPHSKAQLQQT
3391
58.681
0.315
0.465
0.045
0.529
0.333
81.201

KTFNGGGSPHSKAQYQQT
3565
58.609
0.000
0.000
0.000
0.163
0.045
72.618

KSINGSGSPHSKTQSQQT
3392
58.608
0.000
3.017
0.000
0.155
0.017
71.397

KTVNGGGSPHSKAQHQQT
3566
58.602
0.729
0.000
0.000
0.000
0.043
138.544

KSEKGSGSPHSKAQNQQT
3393
58.566
0.000
0.010
0.011
1.601
0.059
158.931

KNVNGSGSPHSKAQNQQT
3394
58.481
0.000
0.000
0.000
0.917
0.166
53.379

KGGEGSGSPHSKAQNQQT
3395
58.472
0.000
0.034
0.000
0.037
0.066
91.023

KTINGSGSPHSKAQRMST
3396
58.435
0.192
0.037
0.000
1.707
0.882
53.414

KTINGSGSPHSKAQGILT
3397
58.418
0.000
0.005
0.010
0.569
0.192
102.631

KEFVGSGSPHSKAQNQQT
3398
58.374
0.000
0.046
0.000
0.088
0.326
128.675

KTIIGSGSPHSKAQDRQT
3399
58.258
1.393
0.230
0.219
0.000
0.045
53.981

KSDKGSGSPHSKAQNQQT
3400
58.248
0.000
0.000
0.000
0.076
0.166
146.566

KTEQVSGSPHSKAQNQQT
3401
58.247
0.000
0.000
0.000
0.000
0.081
88.487

KTEHVSGSPHSKAQNQQT
3402
58.228
0.000
0.024
0.000
0.433
0.141
71.410

KTINGSGSPHSKARDWQT
3403
58.216
0.000
0.005
0.000
0.800
0.259
120.704

KTENASGSPHSKAQNQQT
3404
58.187
0.000
0.038
0.000
0.371
0.129
88.439

KEVQGSGSPHSKAQNQQT
3405
58.125
0.000
0.000
0.000
0.657
0.000
168.220

KTINGSGSPHSKAQNTHD
3406
58.108
0.000
0.027
0.000
0.410
0.126
81.189

KTINGSGSPHSKAPNLQI
3407
58.022
0.000
0.044
0.000
1.548
0.243
55.714

KTINGSGSPHSKAQERST
3408
58.021
0.000
0.01
0.005
0.829
0.409
87.656

KTSNGSGSPHSKAQNYQT
3409
57.894
0.000
0.082
0.000
0.000
0.110
63.681

KTEYISGSPHSKAQNQQT
3410
57.891
0.000
0.000
0.000
0.076
0.075
57.620

KTINGSGSPHSKAQRTCT
3411
57.863
0.000
0.140
0.129
1.855
1.716
90.146

KTINGSGSPHSKAQIGHT
3412
57.769
0.024
0.000
0.000
0.281
0.154
99.262

KNCWGSGSPHSKAQNQQT
3413
57.756
0.000
0.000
0.000
0.000
0.092
59.888

KTINGSGSPHSKAQGAIT
3414
57.627
0.000
0.000
0.000
0.594
0.161
95.696

KTDVNSGSPHSKAQNQQT
3415
57.593
0.000
0.000
0.000
0.000
0.331
66.127

KSDIGSGSPHSKAQNQQT
3416
57.592
0.000
0.000
0.000
0.844
0.128
107.342

KTINGSGSPHSKAQVPPT
3417
57.316
0.000
0.012
0.000
0.257
0.200
90.220

KTINGSGSPHSKAQVQQI
3418
57.308
0.000
1.113
0.000
0.000
0.113
61.957

KTINGSGSPHSKALMRQT
3419
57.234
0.060
0.036
0.100
1.798
0.517
81.332

KTINGSGSPHSKAQYSVT
3420
57.130
0.000
0.093
0.000
1.235
0.302
60.023

KNSIGSGSPHSKAQNQQT
3421
57.101
0.000
0.052
0.000
0.083
0.074
97.381

KTINGSGSPHSKVPNLQT
3422
57.046
0.000
0.029
0.000
0.459
0.082
50.474

KAINGSGSPHSKAQSQQI
3423
56.976
0.000
0.000
0.000
0.000
0.000
57.052

KTINGSGSPHSKAQAITT
3424
56.924
0.000
0.000
0.000
1.239
0.438
75.250

KTINGSGSPHSKAQKTLT
3425
56.844
0.000
0.017
0.009
1.800
1.400
66.415

KTVNGSGSPHSKAQNQWT
3426
56.823
0.000
0.000
0.299
0.000
0.219
69.906

KTINGSGSPHSKAQLHHT
3427
56.815
0.025
0.000
0.010
0.712
0.368
58.418

KTEQTSGSPHSKAQNQQT
3428
56.683
0.000
0.045
0.000
0.792
0.430
59.360

KTINGSGSPHSKAQNIII
3429
56.630
0.000
0.062
0.123
0.099
0.056
76.742

KNSLGSGSPHSKAQNQQT
3430
56.621
0.000
0.028
0.000
0.308
0.162
101.942

KTIPMEGSPHSKAQNQQT
3567
56.560
0.000
0.000
0.000
1.824
0.371
89.951

KTINGSGSPHSKAQGHHT
3431
56.559
0.000
0.000
0.000
0.632
0.117
71.050

KTDRTSGSPHSKAQNQQT
3432
56.466
0.000
0.000
0.000
0.062
0.160
148.498

KTINGSGSPHSKAQSKVT
3433
56.373
0.000
0.050
0.014
1.021
0.390
76.115

KEVVGSGSPHSKAQNQQT
3434
56.371
0.000
0.000
0.000
0.000
0.323
116.964

KTINGSGSPHSKAQLPST
3435
56.238
0.005
4.258
0.001
1.040
0.185
84.918

KTINGSGSPHSKAIGKQT
3436
56.158
0.000
0.000
0.000
0.887
0.088
110.132

KTEPTSGSPHSKAQNQQT
3437
56.134
0.000
0.000
0.000
0.061
0.527
143.397

KTVNGGGSPHSKSQNQQT
3568
56.114
0.116
0.000
0.000
0.000
0.040
170.548

KTINGSGSPHSKAQAIHT
3438
56.047
0.000
0.000
212.32
0.887
0.890
81.908

KTINGSGSPHSKAQHGLT
3439
55.999
0.000
0.000
0.101
1.913
0.244
117.191

KSELGSGSPHSKAQNQQT
3440
55.997
0.000
0.005
0.000
0.881
0.239
120.521

KTINGSGSPHSKAQFMCT
3441
55.916
0.000
0.000
0.000
0.078
0.448
81.959

KTINVSGSPHSKAQGQQT
3442
55.870
0.000
0.191
0.000
0.592
0.040
87.211

KTINGGGSPHSKAQNQMT
3569
55.778
0.000
0.000
0.000
0.866
0.012
73.177

KTVNGSGSPHSKAQHLQT
3443
55.739
0.091
0.036
0.000
0.062
0.409
62.743

KTIRENGSPHSKAQNQQT
3570
55.605
0.000
0.000
0.016
0.131
0.257
95.931

KTINGSGSPHSKTQNHQN
3444
55.551
0.000
0.048
0.000
0.000
0.099
64.846

KTINGSGSPHSKAQPART
3445
55.513
0.000
0.000
0.328
1.294
0.991
127.301

KTVNGSGSPHSKAQSLQT
3446
55.497
0.000
0.060
0.000
0.000
0.143
69.033

KTINGSGSPHSKSQSQLT
3447
55.430
0.000
0.035
0.000
0.050
0.013
125.577

KTINGSASPHSKAHSQQT
3571
55.293
0.000
0.000
0.000
0.000
0.166
66.252

KTWQNSGSPHSKAQNQQT
3448
55.245
0.000
0.000
0.000
0.111
0.265
114.258

KTINGSGSPHSKAQDRQS
3449
55.137
1.146
0.016
0.106
0.644
0.086
55.701

KTINGSGSPHSKAQMPST
3450
54.986
1.691
0.039
0.028
0.450
0.202
114.331

KTNNGGGSPHSKAQNLQT
3572
54.963
0.000
0.000
0.000
0.000
0.089
80.506

KTINGSGSPHSKAQGSLT
3451
54.717
0.000
0.006
0.013
0.480
0.298
142.786

KTEVTSGSPHSKAQNQQT
3452
54.663
0.000
0.000
0.000
0.323
0.185
81.482

KSINGGGSPHSKAQYQQT
3573
54.612
0.000
0.000
0.000
0.105
0.010
65.952

KTVIGSGSPHSKSQNQQT
3453
54.603
0.000
0.000
0.000
0.000
0.106
69.121

KAVNVSGSPHSKAQNQQT
3454
54.586
0.000
0.000
0.000
0.000
0.023
57.835

KTVNGNGSPHSKSQNQQT
3574
54.586
0.000
0.000
0.000
0.256
0.168
95.384

KTDRNSGSPHSKAQNQQT
3455
54.495
0.000
0.000
0.000
0.823
0.241
85.823

KTINGSGSPHSKAQVPAT
3456
54.475
0.000
0.002
0.000
0.782
0.223
137.743

KGVLGSGSPHSKAQNQQT
3457
54.472
0.000
0.007
0.027
0.359
0.189
145.740

KTLNGNGSPHSKAQNLQT
3575
54.458
0.668
0.000
0.000
0.161
0.172
159.134

KAINGSGSPHSKAQDKQT
3458
54.452
0.000
0.000
0.057
0.044
0.223
56.004

KTSNGSGSPHSKAHYQQT
3459
54.414
0.000
0.251
0.000
0.249
0.204
54.162

KTINGSGSPHSKAQVPST
3460
54.366
0.000
1.001
0.000
0.202
0.139
117.223

KTINGSGSSHSKAQNQQT
3576
54.292
1.709
1.870
1.287
1.075
0.458
67.731

KTELRSGSPHSKAQNQQT
3461
54.289
0.000
0.007
0.040
0.790
0.239
57.814

KNINGSGSPHSKAQNHQT
3462
54.248
0.000
0.034
0.000
0.340
0.075
74.979

KTVNGGGSPHSKAQNHQT
3577
54.246
0.375
0.024
0.000
0.000
0.146
67.188

KTINGSGSPHSKARGEQT
3463
54.207
0.025
0.006
0.005
0.309
0.327
128.098

KTINGGGSPHSKAQYQHT
3578
54.188
0.000
0.000
0.000
0.000
0.223
82.256

KTEDLSGSPHSKAQNQQT
3464
54.156
0.000
0.000
0.000
1.193
0.132
70.198

KTINGSGSPHSKAPGQQT
3465
54.071
0.065
0.000
0.004
0.542
0.179
73.440

KTIPKNGSPHSKAQNQQT
3579
53.824
0.000
0.032
0.000
0.115
0.178
77.458

KTINGSGSPHSKAQSLQI
3466
53.778
0.000
0.186
0.000
0.022
0.047
51.543

KTINGSGSPHSKRLEQQT
3467
53.512
0.000
0.118
0.003
0.161
0.292
71.704

KTERGSGSPHSKAQNQQT
3468
53.475
0.000
0.030
0.000
1.416
0.175
85.368

KTVNGSGSPHSKAPNQQT
3469
53.444
0.833
2.206
0.006
0.156
0.178
58.080

KTSNGSGSPHSKAQNQST
3470
53.353
0.000
0.000
0.000
0.000
0.014
120.897

KTINGSGSPHSKAQKVIT
3471
53.273
0.000
0.000
0.000
0.357
0.402
95.147

KTEGISGSPHSKAQNQQT
3472
53.270
0.000
0.000
0.000
0.000
0.010
78.303

KTINGSGSPHSKAQNNDQ
3473
53.226
0.000
0.000
0.000
0.593
0.046
59.664

KTINGSGSPHSKAQSVHT
3474
53.226
0.000
0.004
0.000
0.446
0.217
76.110

KTINGSGSPHSKAQPLGT
3475
53.049
0.015
0.004
0.001
0.515
0.222
68.656

KTINKEGSPHSKAQNQQT
3580
53.006
0.000
0.029
0.000
0.177
0.111
64.520

KTCNASGSPHSKAQNQQT
3476
52.998
0.000
0.011
0.000
0.897
0.141
67.934

KAINGSGSPHSKAHNQET
3477
52.973
0.000
0.030
0.000
0.035
0.058
71.809

KTEGLSGSPHSKAQNQQT
3478
52.891
0.000
0.000
0.020
0.104
0.155
104.529

KTRDASGSPHSKAQNQQT
3479
52.861
0.000
0.000
0.010
1.062
0.402
52.089

KTSNGSGSPHSKAQNLQI
3480
52.843
0.000
0.000
1.605
0.178
0.214
74.823

KTGNGSGSPHSKAQIQQT
3481
52.809
0.000
0.000
0.000
0.000
0.012
98.291

KTVNGGGSPHSKAQNLQT
3581
52.788
0.000
0.031
0.000
0.000
0.165
83.215

KTDRSSGSPHSKAQNQQT
3482
52.737
0.000
0.000
0.000
0.995
0.085
123.421

KTINGSGSPHSKAQVRNT
3483
52.735
0.000
0.101
0.011
0.230
0.423
68.893

KTINGSGSPHSKAPSNQT
3484
52.680
1.494
4.762
0.003
0.330
0.208
87.951

KTINGSGSPHSKAQVGHT
3485
52.624
0.000
0.000
0.006
0.535
0.192
106.448

KNAIGSGSPHSKAQNQQT
3486
52.516
0.000
0.000
0.000
0.165
0.198
117.939

KAENGSGSPHSKAQNQQT
3487
52.487
0.000
0.157
0.029
0.000
0.242
120.256

KTINGSGSPHSKAQRDIT
3488
52.415
0.098
0.000
0.008
1.784
0.605
88.122

KTINGSGSPHSKAQMPNT
3489
52.408
0.084
0.036
0.025
0.057
0.359
66.040

KTVNGSGSPHSKSQNQQT
3490
52.395
0.033
0.077
0.013
0.105
0.175
58.000

KTIPAIGSPHSKAQNQQT
3582
52.346
0.000
0.009
0.000
0.034
0.134
51.949

KTINGSGSPHSKARGLQT
3491
52.275
0.000
0.000
0.036
1.235
1.425
169.881

KTELGSGSPHSKAQNQQT
3492
52.232
0.000
0.007
0.006
0.532
0.088
87.314

KAETGSGSPHSKAQNQQT
3493
52.219
0.000
0.047
0.581
0.009
0.188
132.940

KTINGSGSPHSKLQKQQT
3494
52.144
0.615
0.477
1.071
1.113
0.429
61.833

KTINGSGSPHSKAPSLQT
3495
52.137
0.041
1.614
0.002
0.902
0.222
70.363

KTINGSGSPHSKAQRDQT
3496
51.897
0.069
0.014
0.040
0.867
0.554
102.317

KTDVGSGSPHSKAQNQQT
3497
51.849
0.000
0.007
0.000
0.385
0.560
115.774

KTINGSGSPHSKNRDQQT
3498
51.830
0.000
0.008
0.000
0.480
0.138
100.300

KSINGSGSPHSKAPNLQT
3499
51.812
0.000
0.256
0.000
0.085
0.139
59.270

KTINGSGSPHSKAQAKGT
3500
51.727
0.048
0.016
0.000
0.271
0.525
104.917

KTVNGSGSPHSKAQDKQT
3501
51.580
0.428
0.000
0.069
0.041
0.063
69.225

KTINGGGSPHSKAQNPQA
3583
51.574
0.000
0.000
0.000
0.192
0.000
102.792

KTINGSGSPHSKAQSAHT
3502
51.569
0.068
0.070
0.000
0.589
0.249
79.498

KTINGNGSPHSKSQNQHT
3584
51.379
0.000
0.054
0.000
0.000
0.082
56.614

KTVPTSGSPHSKAQNQQT
3503
51.348
0.013
0.000
0.000
1.017
0.338
102.651

KTIDGSGSPHSKSQNHQT
3504
51.307
0.000
0.000
0.000
0.000
0.269
63.174

KTDVKSGSPHSKAQNQQT
3505
51.296
0.000
0.000
0.000
0.515
0.224
53.601

KAINRSGSPHSKAQDQQT
3506
51.262
0.000
0.000
0.000
0.000
0.036
54.631

KTINGSGSPHSKAQSTMT
3507
51.249
0.018
0.002
0.002
0.321
0.341
73.213

KTVNASGSPHSKAQNQLT
3508
51.249
0.000
0.000
0.000
0.000
0.268
99.559

KTINGSGSPHSKAQREMT
3509
51.076
0.000
24.900
143.49
1.564
0.476
70.961

KTVHGSGSPHSKAQSQQT
3510
51.057
0.000
0.000
0.000
0.143
0.146
54.185

KTINGGGSPHSKSQNRQT
3585
51.017
0.000
0.000
0.000
0.000
0.421
149.370

KTINGSGSPHSKAQYRAT
3511
51.008
0.000
0.158
0.000
0.690
0.120
50.650

KTINGGGSPHSKAQRQQT
3586
50.998
0.000
0.041
0.000
0.991
0.142
147.942

KTEPMSGSPHSKAQNQQT
3512
50.960
0.203
0.000
0.000
1.816
0.415
126.322

KTINGSGSPHSKNQWQQT
3513
50.800
0.000
0.044
0.047
0.111
0.324
65.506

KETAGSGSPHSKAQNQQT
3514
50.762
0.000
0.027
0.000
1.706
0.054
212.795

KTINGSGSPHSKAQRMNT
3515
50.686
0.000
108.747
0.019
0.943
0.264
97.975

KNNLGSGSPHSKAQNQQT
3516
50.670
0.000
0.019
0.000
0.406
0.121
102.408

KTINGSGSPHAKAQNHQT
3517
50.667
0.211
0.140
0.051
0.101
0.090
80.603

KTIIKNGSPHSKAQNQQT
3587
50.587
0.000
0.000
0.000
0.000
0.751
75.547

KTINGSGSPHSYHVNQQT
3588
50.486
0.000
0.056
0.059
0.528
0.275
179.489

KTINGSGSPHSKAGDSQT
3518
50.457
0.614
0.236
0.008
1.062
0.071
74.355

KTINGSGSPHSKLKSQQT
3519
50.368
0.000
0.296
0.000
1.796
1.096
95.240

KTINGSGSPHSKAQKIST
3520
50.285
0.000
0.000
0.088
0.108
0.302
51.115

KTEYNSGSPHSKAQNQQT
3521
50.256
0.000
0.000
0.000
0.000
0.009
62.679

KTINGSGSPHSKAPSMQT
3522
50.249
0.000
0.000
0.004
0.941
0.460
75.504

EAINGSGSPHSKAQNQQT
3523
50.243
0.629
0.094
0.000
0.057
1.519
117.305

KTINGSGSPHSKASPRQT
3524
50.227
0.088
0.005
0.068
1.761
0.530
67.241

KTINGSGSPHSKRMEQQT
3525
50.177
0.000
0.000
0.000
1.327
0.208
81.769

KTINGSGSPHSKAQYQNT
3526
50.099
0.000
0.008
0.000
0.017
0.119
71.846

KTERVSGSPHSKAQNQQT
3589
96.943
0.000
0.000
0.000
0.000
0.144
135.438

KAEIGHDSPHKSGQNQQT
1754
63.249
0.000
0.000
0.000
0.060
0.024
27.173

Table 16 provides the peptide sequence of 341 matured capsid variants, and the fold enrichment of these matured capsid variants relative to the AAV9 control that demonstrated a 75-fold or greater increase in expression in the brain of NHPs relative to the AAV9 control and had a fold-change in expression that was less than 2 relative to the AAV9 control in the liver and the DRG.

TABLE 16

NGS fold-enrichment of TTM-001 and TTM-002 matured AAV capsid variants in the

brain of NHPs

Fold Enrichment relative to AAV9

SEQ

Liver
Liver

ID
Brain
DRG
Heart
Muscle
RNA
DNA
Brain

Sequence
NO:
(NHP)
(NHP)
(NHP)
(NHP)
(NHP)
(NHP)
(Mouse)

KTFNRSGSPHSKAQNQQI
3591
86.359
0.000
113.67
0.000
0.000
0.092
25.568

KTIIGSGSPHSKAQNRHT
3239
217.176
0.000
0.000
0.000
0.000
0.000
210.515

KTFPGSGSPHSKVQNQQT
3240
199.720
0.000
0.000
0.000
0.000
0.967
97.703

KTEKMSGSPHSKAQNQQT
3241
169.461
0.523
0.000
0.000
0.000
0.158
109.161

KAINGHDSPHKSGQIRQT
3606
108.510
0.000
23.908
0.000
0.132
0.261
8.862

KTINGHDSPHKIGQNQHA
3607
77.321
0.000
18.836
0.028
0.220
0.132
7.578

KEINGRGSPHSKAQNQQT
3527
134.390
0.239
0.000
0.000
0.000
0.232
52.311

KTVNRNGSPHSKAQNQQT
3528
133.016
0.000
0.416
0.000
0.000
0.000
85.361

KAINGYDSPHKSGQKQQT
3608
83.803
0.041
9.491
0.000
0.031
0.150
13.057

KTVNGSGSPHSKARDQQT
3242
124.789
0.123
0.039
0.312
0.569
0.454
132.137

KTESGHDSPHKSGQNQQT
3609
86.513
0.000
7.414
0.000
0.000
0.038
13.163

KTINGHDSPHKSGQSVQT
3610
75.748
0.010
6.808
0.000
0.165
0.058
9.321

KTFNGSGSPHSKAPNLQT
3243
121.436
0.000
0.167
0.000
0.000
0.015
168.920

KTEKTSGSPHSKAQNQQT
3244
120.337
0.000
0.355
0.000
0.000
0.119
101.467

TTINGHDSPHKSGQNQQT
3611
108.963
1.512
3.445
0.869
0.659
1.109
14.788

KTINGHESPHKSGRSQQT
3612
97.106
0.000
3.329
0.022
0.000
0.181
9.378

KTINGSGSPHSKAHVRQT
3245
119.798
0.000
0.000
0.262
0.694
1.039
165.590

KTVNGSGSPHSKAPNQHT
3246
117.207
0.000
0.109
0.000
0.000
0.074
51.008

KTEKISGSPHSKAQNQQT
3247
116.603
0.000
0.000
0.000
0.000
0.426
102.978

KTINGPGSPHSKAHNQQT
3529
115.742
0.146
0.000
0.235
0.000
0.513
52.508

KTINGHDSPHKSGQNKLE
3613
76.204
0.000
1.430
0.000
0.015
0.031
12.419

KTVNGSGSPHSKTQSQQT
3248
115.086
0.000
0.726
0.000
0.000
0.340
63.248

TTINGSGSPHSKAQNQQT
3249
114.856
1.340
14.856
0.827
1.281
0.957
72.058

KSINESGSPHSKAQNQQI
3250
113.833
0.000
0.000
0.000
0.000
0.000
67.649

KTINGHDSPHKTGQNQQK
3614
77.562
0.000
1.056
0.000
0.000
0.000
6.379

KTERTSGSPHSKAQNQQT
3251
112.957
0.000
0.009
0.000
1.128
0.207
117.374

KTINGSGSPHSKAQPAKT
3252
111.472
0.331
0.000
1.089
0.044
1.796
215.275

KTINGRGSPHKRGQNQQT
3837
120.889
0.100
0.814
0.434
0.458
0.614
13.988

KTINGSGSPHTKAQNPPT
3592
147.061
0.000
0.727
0.000
0.000
0.000
34.425

KTEKSSGSPHSKAQNQQT
3253
107.470
0.000
0.016
0.014
0.977
0.179
100.177

KAINGHDNPHKSGQNQQT
3615
88.906
0.297
0.721
0.482
0.222
0.130
9.702

KTSYGNGSPHSKAQNQQT
3530
105.937
0.000
0.000
0.000
0.000
0.114
105.894

KTINGQDSPHKSGQHQQA
3616
85.657
1.127
0.579
0.000
0.193
0.557
5.582

KTEKGSGSPHSKAQNQQT
3254
105.614
0.053
0.031
0.000
0.586
0.169
84.653

KTINGSGSPHSKSQTQQN
3255
104.474
0.000
0.131
0.000
0.084
0.038
54.021

KTERISGSPHSKAQNQQT
3256
103.692
0.000
0.000
0.000
0.062
0.370
89.637

KTERASGSPHSKAQNQQT
3257
103.669
0.000
0.000
0.000
0.127
0.070
115.550

KSINGHDSPHKSGQIQHT
3617
87.598
0.000
0.480
0.000
0.714
0.347
13.872

KELHGSGSPHSKAQNQQT
3258
102.680
0.000
0.000
0.000
1.634
0.592
96.554

KAINGSGSPHSKAQNLAT
3259
101.954
0.000
10.954
8.655
0.298
0.239
116.685

KTVNGSGSPHSKSQNQLT
3260
101.327
0.000
0.035
0.000
0.000
0.025
80.716

KAINGHDSPHKSGPRQQT
3618
145.142
0.000
0.408
0.000
0.000
0.000
8.259

KTVNGHDSPHKSGHTQQT
3619
82.246
0.000
0.378
1.142
0.000
0.123
6.160

KSINGHDSPHKSGQRQHT
3620
80.132
0.000
0.357
0.000
0.000
0.000
9.851

KTERNSGSPHSKAQNQQT
3261
99.892
0.000
0.000
0.000
0.000
0.107
87.392

KSLNGSGSPHTKAQNQQT
3593
81.515
0.197
0.333
0.000
0.000
0.085
45.140

KSVNGNGSPHSKAQNQQT
3531
99.385
0.000
1.329
0.000
0.359
0.079
51.016

KAINGHDSPHKSAQSQQT
3621
95.204
0.146
0.310
0.000
0.699
0.058
14.595

KSIYGHESPHKSGQNQQS
3622
90.947
0.817
0.310
0.000
0.000
0.243
8.064

KTFNGSGSPHSKAQGQQT
3262
99.253
0.000
0.208
0.000
0.128
0.099
81.459

KTVNGHDSPHKSLQNQQT
3623
112.925
0.000
0.301
0.059
0.000
0.322
16.726

KTINGSGSPHGWVQNQQT
3532
97.122
0.000
0.000
0.000
1.240
1.975
290.720

KTINGHGSPHSKAQNPQT
3838
83.478
0.000
0.288
0.219
0.000
0.260
11.001

KTSNGYDSPHKSGQKQQT
3624
77.001
0.032
0.286
0.000
0.000
0.016
8.813

KTVNGHDSPHKSGRNQET
3625
102.695
0.000
0.286
0.000
0.000
0.027
11.958

KTTNGHDSPHKSGQTQLT
3626
115.637
0.000
0.283
0.000
0.052
0.321
17.885

KAINGHDSPHKSEKNQQT
3627
77.103
0.000
0.274
0.000
0.000
0.000
26.868

KTERVSGSPHSKAQNQQT
3263
96.943
0.000
0.000
0.000
0.000
0.144
135.438

KTINGSGSPHSKALNRQS
3264
96.843
0.136
0.532
0.000
0.042
0.178
55.945

KTERLSGSPHSKAQNQQT
3265
95.857
0.000
0.004
0.005
0.126
0.260
102.372

KIINGRDSPHKSGQDQQT
3628
78.773
0.000
0.254
0.000
0.000
0.156
16.132

KTDNGSGSPHSKAHNQQT
3266
95.164
0.000
0.000
0.000
0.000
0.027
55.313

KTFHGSGSPHSKTQNQQT
3267
94.714
0.000
0.210
0.120
0.000
0.000
51.119

KTISGHDSPHKTGHNQQT
3629
92.490
0.000
0.233
0.057
0.730
0.000
8.823

KTVNAHDSPHKSGQNQLT
3630
79.137
0.000
0.233
0.178
0.753
0.045
29.254

KTINGGGSPHSKAQTQQI
3533
92.345
0.000
0.000
0.000
0.000
0.023
54.199

KSINGYDSPHKSGQTQQT
3631
79.227
1.817
0.226
0.000
0.000
1.148
4.497

KTINGHESPHKSGQTQQI
3632
86.089
0.000
0.222
0.000
0.000
0.024
3.989

KTINGHDSPHKSGQSKQA
3633
101.460
0.000
0.222
0.000
0.185
0.114
7.510

KTSNGSGSPHSKAQNPPT
3268
91.528
0.000
0.000
0.000
0.000
0.039
51.541

ETINGSGSPHSKAQNLQT
3269
90.969
0.221
1.023
0.197
0.179
0.813
107.216

KTVHGNGSPHSKAQNQQT
3534
90.073
0.000
0.000
0.000
0.000
0.304
97.003

NTINGSGSPHSKAQNQQT
3270
90.017
1.712
1.261
1.171
0.923
0.540
55.179

KTINGGGSPHSKAQNQQC
3535
89.301
0.219
0.000
0.000
0.287
0.319
53.840

KTENMSGSPHSKAQNQQT
3271
89.247
0.000
0.000
0.000
0.000
0.260
130.568

KTENVSGSPHSKAQNQQT
3272
88.506
0.000
0.000
0.000
0.964
0.112
108.591

KTSSGSGSPHSKAQYQQT
3273
87.304
0.000
0.000
0.000
0.000
0.299
58.143

KTIDGGGSPHSKAQNKQT
3536
85.019
0.000
0.000
0.000
0.000
0.477
55.517

KTEKVSGSPHSKAQNQQT
3274
84.558
0.000
0.022
0.000
0.873
0.424
112.185

KAINGSGSPHSKAQDQET
3275
84.080
0.000
0.000
0.000
0.194
0.027
87.637

KTCNKSGSPHSKAQNQQT
3276
83.992
0.000
0.000
0.165
0.283
0.000
119.496

KTINGGGSPHSKAQNQLI
3537
83.881
0.000
0.000
0.000
0.046
0.387
78.383

KNINGGGSPHSKAQNQQT
3538
83.083
0.000
0.042
0.000
0.000
0.000
75.913

KTEHLSGSPHSKAQNQQT
3277
83.080
0.000
0.000
0.012
0.021
0.189
69.494

KAIIGHESPHKSGQNQQT
3634
88.563
0.000
0.150
0.000
0.062
0.145
8.530

KTINGHDSPHKTGQNQPP
3635
77.357
0.000
0.149
0.000
0.000
0.096
8.865

KAINGHDSPHKSGQSPQT
3636
75.734
0.095
0.148
0.000
0.000
0.238
14.195

KAEMGSGSPHSKAQNQQT
3278
83.049
0.000
0.020
0.000
0.768
0.112
135.019

KATNGSGSPHSKAQNHQT
3279
82.627
0.000
0.176
0.000
0.155
0.057
66.207

KTIKGNDSPHKSVQNQQT
3637
85.986
0.000
0.135
0.000
0.263
0.000
8.603

KAIKGSGSPHSKAQDQQT
3280
82.258
0.000
0.000
0.000
0.108
0.000
85.178

KTINGGGSPHSKSQNQLT
3539
82.231
0.000
0.070
0.000
0.000
0.498
126.986

KTEFGHDSPHKSGQNQQT
3638
77.245
0.000
0.124
0.000
0.561
0.063
16.337

KTINGHDSPHKSAQNYQT
3639
130.375
0.000
0.124
0.000
0.097
0.123
19.443

KTFNGSASPHSKALNQQT
3839
84.258
0.000
0.122
0.000
0.104
0.037
31.855

KTINGCGSPHASGQNQQT
3840
132.540
0.000
0.121
0.042
0.000
0.059
1.857

KTINAHDSPHKIGQNHQT
3640
106.832
0.000
0.121
0.000
0.000
0.231
5.074

KTVNGNGSPHSKAQNKQT
3540
81.481
0.000
0.000
0.000
0.000
0.122
69.455

KTINGHESPHKSAQNRQT
3641
95.531
0.000
0.113
0.000
0.130
0.082
4.815

KTINGSGSPHSKGHWQQT
3281
81.434
0.000
0.000
0.000
0.000
1.011
65.252

KTTNGHDSPHKSGQNQQG
3642
85.113
0.000
0.107
0.000
0.000
0.017
10.555

KTIKGQDSPHKIGQNQQT
3643
110.357
0.000
0.103
0.058
0.166
0.135
11.829

KTDKTSGSPHSKAQNQQT
3282
81.430
0.000
0.000
0.000
1.362
0.291
169.515

KTVNGHDSPHKSGQNHLT
3644
81.516
0.000
0.100
0.017
0.000
0.028
16.096

KTFKGSGSPHSKAPNQQT
3283
80.890
0.000
0.000
0.000
0.000
0.017
71.144

KSINGHDSPHKSGQYQHT
3645
88.195
0.000
0.099
0.000
0.000
0.149
14.485

KTINGNDSPHKSVQNHQT
3646
120.002
0.000
0.099
0.788
0.000
0.000
7.920

KTVNGSGSPHSKAQNQLI
3284
80.509
0.000
0.000
0.000
0.000
0.166
71.156

KTINGSGSPHSKRPEQQT
3285
80.418
0.000
0.013
0.000
0.149
0.361
50.319

KTINGSGSPHSKAQRTMT
3286
80.388
0.000
0.022
0.170
1.812
1.025
100.248

KTITGHDSPHKSGQNQWT
3647
81.658
0.000
0.090
0.000
0.936
0.000
7.744

KTNNGHDSPHKSVQNQHT
3648
115.172
0.000
0.083
0.000
0.000
0.062
7.934

KTEKASGSPHSKAQNQQT
3287
80.285
0.000
0.041
0.000
0.000
0.261
90.390

KTIDGHDSPHKSGQNQHA
3649
91.058
0.000
0.082
0.000
0.000
0.000
10.781

KSDQGSGSPHSKAQNQQT
3288
80.076
0.000
0.000
0.000
0.993
0.124
151.911

KTVNGHDSPHKSGQTRQT
3650
133.276
0.251
0.080
0.093
0.034
0.129
7.174

KTVNGHDSPHKSGQNLHT
3651
88.080
0.000
0.080
0.000
0.000
0.039
11.363

KAISGHDSPHKSGLNQQT
3652
78.846
0.000
0.079
0.000
0.000
0.015
11.045

KTEITSGSPHSKAQNQQT
3289
79.620
0.000
0.163
0.000
0.332
0.074
76.686

KTDKSSGSPHSKAQNQQT
3290
79.470
0.055
0.012
0.000
1.437
0.367
141.351

KAINGHDSPHKSAQNQET
3653
90.402
0.000
0.073
0.000
0.746
0.000
10.674

KTITGHDSPHKSGQHLQT
3654
137.945
0.000
0.072
0.000
0.000
0.000
4.187

KTIDGSGSPHSKAQNQQH
3291
79.090
0.000
0.000
0.000
0.136
0.049
57.914

KTVNGNGSPHSKAQNQHT
3541
78.849
0.000
0.000
0.000
0.000
0.045
54.086

KNINGSGSPHSKAQNQQT
3292
78.445
0.000
0.000
0.000
0.571
0.177
89.719

KTINGHDSPHKSRLNQPT
3655
92.883
0.000
0.070
0.050
0.904
1.075
5.598

KTETHSGSPHSKAQNQQT
3293
77.974
0.000
0.067
0.000
0.000
0.512
57.287

KTVDGHDSPHKSGQKQQT
3656
78.802
0.000
0.069
0.000
0.157
0.342
7.200

KTINGQDSPHKSGQNQDT
3657
82.075
0.000
0.067
0.000
0.225
0.144
9.626

KTINGGGSPHSKALNQQN
3542
77.822
0.000
0.131
0.000
0.000
0.274
69.884

KTIEGHDSPHKSGRNQQT
3658
75.838
0.000
0.065
0.017
0.000
0.079
7.818

KTTNGHDSPHKSGQNLLT
3659
77.738
0.130
0.064
0.185
0.424
0.326
15.192

KTINGHDSPHKSGQLVIT
3660
76.781
0.089
0.064
0.000
0.338
0.475
11.323

KTVNGHDSPHKSRQSQQT
3661
76.458
0.000
0.063
0.000
0.000
0.021
8.136

KTINGSGSPHSKALHQHT
3294
77.502
0.000
0.052
0.041
0.000
0.188
68.196

KTINGHDSPHKSGRTQET
3662
81.599
0.000
0.062
0.000
0.000
0.137
7.270

KTINGHDSPHKSVQTHQT
3663
77.309
0.237
0.062
0.000
0.000
0.116
7.519

KTINGTGSPHSKAQNHQI
3543
77.089
0.171
0.000
0.000
0.000
0.166
54.281

KTINGSGSPHSKAQHRIT
3295
76.849
0.105
0.499
0.170
1.424
0.214
127.000

KTINGSGSPHSKAQYIHT
3296
76.170
0.000
0.014
0.033
1.523
0.168
59.649

KTSNGHDSPHKSGQNQPA
3664
75.834
0.000
0.056
0.000
0.000
0.000
8.501

KTEGKHDSPHKSGQNQQT
3665
98.384
0.000
0.056
0.000
0.000
0.000
10.345

KTENISGSPHSKAQNQQT
3297
76.072
0.000
0.000
0.000
0.115
0.132
83.118

KVINGHDSPHKSGQTQQT
3666
91.665
0.000
0.055
1.526
0.311
0.000
7.391

KTIIGGGSPHSKAHNQQT
3544
75.872
0.000
0.050
0.000
0.000
0.235
65.492

KTINGPDSPHKIGQNQQS
3667
85.726
0.000
0.055
0.171
0.000
0.063
10.055

KTINGSGSPHSKAQKFET
3298
75.788
0.000
0.000
0.028
0.108
0.093
65.588

KTSNESGSPHSKAQNHQT
3299
75.720
0.000
0.000
0.000
0.169
0.217
70.590

KTINGSGSPHSKAQFPST
3300
75.677
0.000
0.004
0.000
0.849
0.127
119.712

KTERPSGSPHSKAQNQQT
3301
75.669
0.000
0.029
0.000
0.000
0.156
73.894

KAVNGHDSPHKSVQNQQT
3668
81.051
0.448
0.051
0.000
0.665
0.091
11.288

KTINGNGSPHSKAQNPLT
3545
75.269
0.000
0.000
0.000
0.366
0.000
53.583

KSIKGNGSPHSKAQNQQT
3546
75.196
0.000
0.000
0.000
0.000
0.000
90.251

KTINGHDSPHKSRQDQHT
3669
75.595
0.000
0.049
0.118
0.030
0.045
8.540

KAINGPDSPHKSGQKQQT
3670
78.213
0.464
0.047
0.000
0.323
0.162
10.395

KTINGHDSPHKSRQSQHT
3671
88.544
0.499
0.046
0.000
0.059
0.032
8.324

KTIYGHDSPHKSVQNQLT
3672
92.381
0.000
0.043
0.000
0.103
0.016
12.323

KTVNGHDSPHKSGQNLLT
3673
83.969
0.114
0.040
0.023
0.000
0.035
18.894

KTESAHDSPHKSGQNQQT
3674
80.810
0.000
0.039
0.000
0.000
0.000
13.338

KTENKSGSPHSKAQNQQT
3594
103.854
0.000
0.037
0.000
0.000
0.119
31.182

KTTNGQDSPHKSGQNQQS
3675
92.419
0.000
0.037
0.043
0.000
0.079
7.592

KTDKGSGSPHSKAQNQQT
3595
94.572
0.000
0.037
0.000
0.951
0.367
47.888

KTIDGHDSPHKSGRNQQI
3676
80.240
0.000
0.037
0.000
0.040
0.144
10.363

KTINGYDSPHKSGQYQHT
3677
81.534
0.000
0.036
0.000
0.000
0.000
10.524

KTDNGHDSPHKSRQNQQT
3678
105.312
0.000
0.033
0.000
0.000
0.018
7.931

KTINGHDSPHKSWVRQQT
3679
125.537
0.000
0.033
0.000
0.291
0.174
11.687

KTINGHESPHKSGQNQHS
3680
92.248
0.000
0.032
0.012
0.090
0.088
9.720

KTVNGHDSPHKIGHNQQT
3681
120.985
0.000
0.029
0.000
0.000
0.009
10.167

KTCNGHDSPHKSGRNQQT
3682
94.616
0.000
0.025
0.000
0.000
0.128
12.496

KTINGNGSPHSKAQNHQA
3841
88.274
0.000
0.024
0.000
0.000
0.041
36.754

KNVVGHDSPHKSGQNQQT
3683
75.330
0.000
0.024
0.000
0.063
0.049
8.077

KTELWHDSPHKSGQNQQT
3684
85.323
0.057
0.020
0.000
0.000
0.243
9.915

KTELRHDSPHKSGQNQQT
3685
98.098
0.000
0.019
0.000
0.000
0.007
6.588

KTINGHDSPHKSNAWQQT
3686
84.825
0.000
0.016
0.000
0.000
0.132
15.788

KTDAGHDSPHKSGQNQQT
3687
88.924
0.000
0.013
0.000
1.076
0.070
18.107

KTEVGHDSPHKSGQNQQT
3688
112.457
0.000
0.011
0.000
0.000
0.138
13.125

KTESRHDSPHKSGQNQQT
3689
81.766
0.000
0.011
0.000
0.052
0.036
6.975

KSELGHDSPHKSGQNQQT
3690
107.059
0.000
0.005
0.000
0.000
0.055
13.285

KTINGHDSPHKSGQSVPT
3691
77.840
0.000
0.003
0.000
0.136
0.061
6.768

KTINGHESPHKSGQNIQP
3692
253.840
0.000
0.000
0.000
0.000
0.000
14.042

KTEMKHDSPHKSGQNQQT
3693
240.075
0.000
0.000
0.000
0.000
0.000
3.183

KTINGHDSPHKSVQNHLN
3694
196.758
0.000
0.000
0.000
0.000
0.000
14.557

KTINGHDSPHKIGLDQQT
3695
165.627
0.000
0.000
0.000
1.942
0.000
5.469

KTSNASGSPHSKAQHQQT
3596
165.206
0.000
0.000
0.000
0.000
0.082
40.558

KTINGHDSPHKRGPDQQS
3696
160.084
0.000
0.000
0.000
0.000
0.000
2.923

KTINGMGSPHSKTQNQQT
3842
158.728
0.000
0.000
0.000
0.000
0.638
47.809

KTIKGHDSPHKSGESQQT
3697
142.264
0.000
0.000
0.000
0.000
0.218
4.176

KTEGWHDSPHKSGQNQQT
3698
142.064
0.000
0.000
0.000
0.000
0.264
11.785

KTINGHDSPHKHGQNHQT
3699
141.405
0.191
0.000
0.000
0.000
0.000
10.214

KTEQLHDSPHKSGQNQQT
3700
138.345
0.000
0.000
0.000
0.000
0.000
12.606

KTVNGTGSPHSKAQNQLT
3843
137.639
0.000
0.000
0.000
0.000
0.277
48.950

KTIIGHDSPHKSGQYQHT
3701
131.825
0.000
0.000
0.000
0.000
0.211
5.762

KTSNGHDSPHKSVQNKQT
3702
130.640
0.000
0.000
0.000
0.172
0.039
11.850

KIVNGQVSPHKSGQNQQT
3703
129.649
0.000
0.000
0.000
0.000
0.031
16.942

KTVNGHDSPHKSGQRQLT
3704
129.641
0.000
0.000
0.000
0.000
0.487
20.145

KTVNGHDSPHKIGQNQLT
3705
128.582
0.000
0.000
0.499
0.027
0.199
20.957

KTINGHDSPHKSGQIIVT
3706
125.245
0.000
0.000
0.151
0.000
0.379
6.808

KTEKIHDSPHKSGQNQQT
3707
125.178
0.000
0.000
0.000
0.000
0.000
17.604

KTENAHDSPHKSGQNQQT
3708
124.477
0.000
0.000
0.000
0.000
0.062
15.805

KIGNGHESPHKSGQNQQT
3709
123.324
0.000
0.000
0.000
0.000
0.000
11.198

KEVMGHDSPHKSGQNQQT
3710
121.107
0.000
0.000
0.000
0.000
0.000
17.191

KTEVKHDSPHKSGQNQQT
3711
119.733
0.000
0.000
0.000
0.000
0.000
5.550

KTINGYDSPHKSGQKQST
3712
119.615
0.000
0.000
0.000
0.000
0.000
7.970

KTIHGNGSPHSKAQNQET
3844
117.388
0.000
0.000
0.000
0.000
0.000
38.874

KYQVGHDSPHKSGQNQQT
3713
112.797
0.000
0.000
0.000
0.000
0.542
9.335

KTEAMHDSPHKSGQNQQT
3714
111.765
0.000
0.000
0.000
0.000
0.000
16.142

KTIKGDDSPHKSVQNQQT
3715
109.397
0.000
0.000
0.000
0.000
0.000
19.125

KTINGHDSPHKSVQSHQT
3716
109.375
0.107
0.000
0.319
0.000
0.547
12.617

KTINGHDSPHKSGQFVVT
3717
108.725
0.000
0.000
0.000
0.124
0.406
10.179

KTVNGHDSPHKSRQNLQT
3718
107.496
0.205
0.000
0.000
1.934
0.062
8.616

KATNGHNSPHKSGQNQET
3719
106.806
0.000
0.000
0.000
0.000
0.000
10.566

KAINGHDSPHKSAQNQQI
3720
106.539
0.000
0.000
0.000
0.000
0.113
21.786

KTEHGHDSPHKSGQNQQT
3721
106.486
0.000
0.000
0.000
0.000
0.006
14.956

KTVENHDSPHKSGQNQQT
3722
106.468
0.000
0.000
0.000
0.000
0.156
9.246

KTIYGHDSPHKSGQSQPT
3723
106.431
0.000
0.000
0.000
0.155
0.137
6.562

KTISGHESPHKSGQNEQT
3724
105.740
0.000
0.000
0.000
0.378
1.384
9.156

KAIIGHDSPHKSAQNQQT
3725
105.292
0.000
0.000
0.000
0.000
0.553
16.793

KAIDGHDSPHKSGQNQLT
3726
104.701
0.331
0.000
0.000
0.201
0.638
16.109

KTIMGHDSPHKSVQNQQT
3727
104.683
0.000
0.000
0.000
0.000
0.000
8.029

KEVGGHDSPHKSGQNQQT
3728
103.896
0.000
0.000
0.000
0.000
0.000
16.899

KTINGHDSPHKSAQNLLT
3729
103.332
0.000
0.000
0.256
0.194
0.000
15.722

KTEFTHDSPHKSGQNQQT
3730
102.052
0.062
0.000
0.000
0.439
0.047
12.527

KTINASGSPHSKAINQQT
3597
101.122
0.233
0.000
0.000
0.000
0.145
47.196

KAINGNGSPHKRGQNQQT
3845
100.925
0.000
0.000
0.000
0.000
0.159
10.011

KSEMGHDSPHKSGQNQQT
3731
100.539
0.000
0.000
0.000
0.000
0.000
18.356

KAQQGHDSPHKSGQNQQT
3732
100.395
0.000
0.000
0.000
0.000
0.057
3.954

KTEVMHDSPHKSGQNQQT
3733
99.473
0.000
0.000
0.000
0.000
0.000
12.400

KAINGHDSPHKSGQSLQT
3734
99.310
0.058
0.000
1.439
0.254
0.056
17.323

KTINGSGSPHSKAPNQQH
3598
99.300
0.252
0.000
0.000
0.000
0.038
39.297

KCGEGHDSPHKSGQNQQT
3735
99.298
0.000
0.000
0.000
0.000
0.000
13.147

KTVNGHDSPHKSAQNHQT
3736
99.257
0.000
0.000
0.078
0.000
0.027
17.639

KTVNGHDSPHKSGQTQLT
3737
98.524
0.000
0.000
0.313
0.183
0.172
14.883

KTNNGHDSPHKSGRNRQT
3738
98.307
0.000
0.000
0.124
0.000
0.037
5.840

KTCNEHDSPHKSGQNQQT
3739
97.092
0.000
0.000
0.000
0.000
0.000
8.364

KTINGHDSPHKYGQNEQT
3740
96.960
0.000
0.000
0.000
0.000
0.000
4.613

KASNRHDSPHKSGHNQQT
3741
96.283
0.000
0.000
0.000
0.000
0.675
8.045

KTINGNGSPHSKAPNLQT
3846
95.963
0.000
0.000
0.000
0.000
0.247
36.341

KTETKHDSPHKSGQNQQT
3742
95.121
0.000
0.000
0.000
0.000
0.083
4.831

KSINGHDSPHKSQQNQQT
3743
94.479
0.000
0.000
0.000
0.000
1.696
9.633

KTIGGHDSPHKSGQNQQI
3744
94.420
0.000
0.000
0.000
0.000
0.333
19.324

KTDPQHDSPHKSGQNQQT
3745
93.931
0.000
0.000
0.000
0.906
0.019
11.749

KTINRHDSPHKIVQNQQT
3746
93.409
0.000
0.000
0.000
0.000
0.000
3.064

KTEQYHDSPHKSGQNQQT
3747
93.065
0.000
0.000
0.000
0.000
0.047
15.190

KTINGHDSPHKSVQSKQT
3748
92.445
0.000
0.000
0.078
0.000
0.047
4.263

KELVGHDSPHKSGQNQQT
3749
92.262
0.000
0.000
0.000
0.594
0.000
14.890

KTENRHDSPHKSGQNQQT
3750
91.675
0.000
0.000
0.000
0.000
0.000
13.282

KELMGHDSPHKSGQNQQT
3751
91.191
0.000
0.000
0.000
0.000
0.000
14.032

KTINGNDSPHKIGHNQQT
3752
91.183
0.000
0.000
0.117
0.000
0.270
11.095

KTIKGGGSPHSKAQDQQT
3847
91.172
0.000
0.000
0.000
0.064
0.085
49.580

KTEGHHDSPHKSGQNQQT
3753
89.922
0.000
0.000
0.000
0.000
0.000
23.929

KTEGYHDSPHKSGQNQQT
3754
89.891
0.000
0.000
0.000
0.000
0.000
15.116

KTVNGHDSPHKSGQTQQI
3755
89.801
0.000
0.000
0.657
0.000
0.542
11.543

KTINGQDSPHKSGQNPLT
3756
89.726
0.000
0.000
0.000
0.363
0.000
15.561

KTVNASGSPHSKAQNHQT
3599
89.467
0.091
0.000
0.000
0.045
0.252
39.024

KTINGHDSPHKSGRDQKT
3757
88.871
0.000
0.000
0.000
0.350
0.181
12.117

KTINGHDSPHKSVHNQQN
3758
88.715
0.089
0.000
0.081
0.000
0.143
10.787

KTINGHDSPHKSGQWKRT
3759
88.633
0.000
0.000
0.000
0.202
0.094
5.186

KTIDGSGSPHSKAENRQT
3600
87.993
0.092
0.000
0.000
0.139
0.054
40.629

KNEIGHDSPHKSGQNQQT
3760
87.758
0.000
0.000
0.000
0.000
0.055
14.110

KAINGHDSPHKSGQSQQI
3761
87.585
0.000
0.000
5.310
0.000
0.000
12.864

KIINGHDSPHKSRQAQQT
3762
86.966
0.000
0.000
0.000
0.000
0.000
9.193

KTPNGHDSPHKSGQNQQI
3763
86.683
0.000
0.000
0.000
0.000
0.109
21.278

KITNGHDSPHKSGQTQQT
3764
86.443
0.000
0.000
0.000
0.192
0.190
17.479

KTINGHDSPHKSVQNHQI
3765
86.395
0.000
0.000
0.000
0.000
0.000
9.148

KTINGHDSPHKSKQNQQA
3766
86.265
0.000
0.000
0.000
0.123
0.041
5.768

KTINGHDSPHKSAQNQLN
3767
86.153
0.000
0.000
0.000
0.050
0.019
15.587

KTDITHDSPHKSGQNQQT
3768
85.876
0.000
0.000
0.000
0.000
0.013
9.076

KTVNGHDSPHKSGQTQPT
3769
85.680
0.000
0.000
1.301
1.064
0.000
8.067

KTEKFHDSPHKSGQNQQT
3770
85.358
0.000
0.000
0.000
0.000
0.026
7.229

KTDQGHDSPHKSGQNQQT
3771
85.267
0.000
0.000
0.000
0.000
0.000
16.042

KTINGHDSPHKLWINQQT
3772
85.132
0.000
0.000
1.154
0.000
0.017
12.704

KGINGPDSPHKSGQNQQT
3773
85.080
0.000
0.000
0.084
0.000
0.054
13.750

KSEIGHDSPHKSGQNQQT
3774
84.789
0.000
0.000
0.000
0.000
0.013
15.955

KTINGHDSPHKSVQKQLT
3775
84.351
0.000
0.000
0.000
0.038
0.103
11.890

KTINGHPSPHWKGQNQQT
3848
84.153
0.000
0.000
0.000
0.000
0.058
3.280

KTVNGHDSPHKSGRNQLA
3776
83.858
0.000
0.000
0.000
0.000
0.132
21.252

KTNNVHDSPHKSGQNQQS
3777
83.697
0.000
0.000
0.000
0.176
0.000
7.117

KTIKGSGSPHSKVQDQQT
3601
83.077
0.000
0.000
0.034
0.000
0.107
21.001

KSEKGHDSPHKSGQNQQT
3778
82.982
0.000
0.000
0.000
0.000
0.105
16.662

KWSAGHDSPHKSGQNQQT
3779
82.949
0.000
0.000
0.000
0.000
0.211
12.499

KELAGHDSPHKSGQNQQT
3780
82.876
0.000
0.000
0.000
0.000
0.093
18.063

KTINGHDSPHKMGRNQQS
3781
82.787
0.000
0.000
0.000
0.000
0.000
6.467

KTDQAHDSPHKSGQNQQT
3782
82.402
0.000
0.000
0.141
0.000
0.000
13.397

KTETQHDSPHKSGQNQQT
3783
82.316
0.000
0.000
0.000
0.000
0.198
10.823

KTEMTHDSPHKSGQNQQT
3784
82.221
0.000
0.000
0.000
0.000
0.000
8.431

KTINGHDSPHKSGISIQT
3785
82.019
0.000
0.000
0.000
0.191
0.044
7.310

KTDAVHDSPHKSGQNQQT
3786
81.968
0.000
0.000
0.000
0.297
0.107
13.596

KTSNGHDSPHKSVQNLQT
3787
81.921
0.000
0.000
0.072
0.000
0.330
11.544

KTEKYHDSPHKSGQNQQT
3788
81.637
0.000
0.000
0.000
0.000
0.013
7.580

KQTQGHDSPHKSGQNQQT
3789
81.581
0.000
0.000
0.000
0.000
0.133
15.225

KTINGHDSPHKMAHNQQT
3790
81.329
0.000
0.000
0.000
0.000
0.094
15.949

KAINGSGSPHSKAQTQQA
3602
81.207
0.000
0.000
0.000
0.000
0.016
40.435

KTINGHDSPHKHGQNQQN
3791
81.065
0.000
0.000
0.000
0.000
0.000
4.110

KGADGHDSPHKSGQNQQT
3792
80.981
0.000
0.000
0.000
0.000
0.074
11.423

KVGEGHDSPHKSGQNQQT
3793
80.775
0.084
0.000
0.000
0.000
0.019
16.378

KANEGHDSPHKSGQNQQT
3794
80.470
0.000
0.000
0.000
0.000
0.000
12.818

KTDTMHDSPHKSGQNQQT
3795
80.364
0.000
0.000
0.000
0.000
0.000
13.166

KTEAKSGSPHSKAQNQQT
3603
80.088
0.192
0.000
0.000
0.000
0.613
47.130

KTINGHDSPHKSVQSQQS
3796
80.000
0.000
0.000
0.000
1.055
0.082
17.620

KTIPGSGSPHSKAQNLQT
3604
79.973
0.871
0.000
0.000
0.000
0.000
32.693

KTCIAHDSPHKSGQNQQT
3797
79.857
0.000
0.000
0.066
0.000
0.093
1.930

KTINGHDSPHKSGQTVCT
3798
79.730
0.000
0.000
0.000
0.050
0.030
7.873

KELRGHDSPHKSGQNQQT
3799
79.596
0.000
0.000
0.000
0.000
0.006
22.001

KCQIGHDSPHKSGQNQQT
3800
79.359
0.000
0.000
0.000
0.000
0.000
2.614

KGVMGHDSPHKSGQNQQT
3801
79.170
0.000
0.000
0.000
0.138
0.086
17.287

KACDGHDSPHKSGQNQQT
3802
78.648
0.000
0.000
0.000
0.000
0.128
17.767

KTINGQDSPHKSGQYQQI
3803
78.585
0.000
0.000
0.000
0.286
0.672
5.664

KTINGHDSPHKSGQQIMT
3804
78.534
0.000
0.000
0.000
0.000
0.058
7.067

KTINGHDSPHKSRQNEQS
3805
78.534
0.000
0.000
0.000
0.112
0.188
13.388

KASNGHDSPHKSGLNHQT
3806
78.451
0.000
0.000
0.000
0.000
0.000
17.975

KTVNGHDSPHKSGQSQPT
3807
78.309
0.000
0.000
0.000
0.000
0.231
10.627

KNELGHDSPHKSGQNQQT
3808
78.135
0.000
0.000
0.000
0.000
0.182
17.457

KTETFHDSPHKSGQNQQT
3809
78.070
0.000
0.000
0.000
0.782
0.007
4.693

KAAEGHDSPHKSGQNQQT
3810
77.793
0.000
0.000
0.000
0.000
0.060
13.552

KGQNGHDSPHKSGQNQQT
3811
77.770
0.000
0.000
0.000
0.107
0.056
13.618

KNEFGHDSPHKSGQNQQT
3812
77.740
0.000
0.000
0.000
0.000
0.029
16.318

KTSIGYDSPHKSGQNQQT
3813
77.730
0.000
0.000
0.000
0.057
0.178
4.831

KTDNGHDSPHKSGQNLQT
3814
77.565
0.504
0.000
0.000
0.000
0.000
16.184

KTEGQHDSPHKSGQNQQT
3815
77.423
0.000
0.000
0.000
0.000
0.748
20.310

KTITGHDSPHKSRQDQQT
3816
77.127
0.000
0.000
0.000
0.000
0.000
6.250

KAEHGHDSPHKSGQNQQT
3817
77.026
0.000
0.000
0.000
0.000
0.017
20.937

KTINGDDSPHKSGQKQLT
3818
76.968
0.000
0.000
0.000
0.163
0.014
15.820

KCDQGHDSPHKSGQNQQT
3819
76.887
0.000
0.000
0.000
0.193
0.013
27.317

KEILGHDSPHKSGQNQQT
3820
76.770
0.000
0.000
0.000
0.804
0.009
10.771

KTIHGSGSPHSKAQNQAT
3605
76.765
0.000
0.000
0.000
0.000
0.215
43.969

KTERNHDSPHKSGQNQQT
3821
76.751
0.000
0.000
0.000
0.000
0.000
14.979

KAINGDDSPHKSGHNQQT
3822
76.578
0.000
0.000
0.000
0.032
0.059
17.755

KTSNGHNSPHKSGQNQET
3823
76.515
0.000
0.000
0.000
0.000
0.000
4.764

KTINGHDSPHKSGQMIHT
3824
76.364
0.000
0.000
0.000
0.000
0.000
9.486

KNAIGHDSPHKSGQNQQT
3825
76.289
0.000
0.000
0.000
0.009
0.072
15.178

KTDKFHDSPHKSGQNQQT
3826
76.204
0.000
0.000
0.000
0.000
0.000
7.096

KTEGFHDSPHKSGQNQQT
3827
76.191
0.000
0.000
0.000
0.000
0.080
13.163

KVINGHDSPHKSGRNHQS
3828
75.961
0.000
0.000
0.000
0.000
0.000
13.568

KTITGHDSPHKSVQNRQT
3829
75.940
0.000
0.000
0.000
0.621
0.000
4.310

KTPDMHDSPHKSGQNQQT
3830
75.871
0.659
0.000
0.000
0.000
0.048
11.277

KTINGHDSPHKSGQKMNT
3831
75.820
0.000
0.000
0.000
0.000
0.167
6.373

KTELQHDSPHKSGQNQQT
3832
75.814
0.000
0.000
0.000
0.105
0.000
11.798

KTIHGHDSPHKSGQSQQN
3833
75.777
0.000
0.000
0.059
0.000
0.166
7.426

KTEIGHDSPHKSGQNQQT
3834
75.525
0.000
0.000
0.016
0.012
0.000
9.593

KTINGHDSPHKSGQYQHA
3835
75.308
0.000
0.000
0.000
0.000
0.017
17.081

KTELYHDSPHKSGQNQQT
3836
75.235
0.000
0.000
0.000
0.000
0.042
10.354

Table 17 provides the sequences of 216 matured capsid variants having a CV of less than 1 for the liver RNA samples isolated and a 10-fold or greater increase in expression relative to AAV9 in the liver of NHPs. These matured variants showed preferential transduction of the liver over other tissues as shown by a low value for fold-enrichment relative to AAV9 in the other tissues investigated including the brain, DRG, heart and muscle. As such, Table 17 provides TTM-001 and TTM-002 matured AAV capsid variants with liver-specific tropism. Across the peptides within the matured capsid variants in Table 17, approximately 175 of them comprised the sequence GSGSPH (SEQ ID NO: 4695) and further comprised additional modifications in the C-terminal region of the sequence.

TABLE 17

NGS fold-enrichment of TTM-001 and TTM-002 matured AAV capsid variants in the

liver of NHPs

Fold Enrichment relative to AAV9

SEQ
Liver
Liver

ID
RNA
DNA
Brain
DRG
Heart
Muscle

Sequence
NO:
(NHP)
(NHP)
(NHP)
(NHP)
(NHP)
(NHP)

KTQRKSGSPHSKAQNQQT
4011
119.659
1.439
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQARKT
4681
96.557
3.644
0.000
0.000
0.000
0.000

KYIVGSGSPHSKAQNQQT
4682
94.721
4.480
0.000
0.000
0.000
0.000

KTINGSGSPHSMYMNQQT
4683
81.106
5.840
0.000
0.000
0.000
0.000

KTINGSGSPHSKAFYRQT
4684
77.541
3.577
0.000
0.000
0.000
0.000

KTINGSGSPHSKLKRQQT
4685
76.103
6.884
0.000
0.000
0.000
0.000

KTINGSGSPHSKRHRQQT
4686
73.225
4.648
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQKCIT
4687
69.547
1.887
0.000
0.000
0.000
0.000

KTINGSGSPHSKWRLQQT
4688
68.083
2.037
0.000
0.000
0.000
0.000

KTINGSGSPHSRCRNQQT
4689
64.416
5.150
0.000
0.000
0.000
0.000

KTINGSGSPHSFTCNQQT
4690
63.936
2.155
0.000
0.000
0.000
271.289

KTINGSGSPHSKFFIQQT
4691
63.255
6.916
0.000
0.000
0.000
0.000

KTINGSYSPHCLAQNQQT
4012
62.942
0.309
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQYSRT
4692
60.119
0.956
0.000
0.000
0.000
0.000

KTINGSGSPHIVWQNQQT
3849
58.021
6.056
0.000
0.000
0.000
0.000

KTINGSGSPHSKYFMQQT
3850
57.350
2.993
0.000
0.000
0.000
0.000

KTINGSGSPHSKARQRQT
3851
56.775
2.205
0.000
0.000
0.000
0.000

KTINGSGSPHSCHQNQQT
3852
56.242
8.562
0.000
0.000
0.000
0.000

KTINGSGSPHFPWQNQQT
3853
53.587
1.731
0.000
0.000
0.000
0.000

KTINGSGSPHSKIRRQQT
3854
53.528
1.388
0.000
0.000
0.000
0.000

KTINGSGSPHVYYQNQQT
3855
53.294
2.173
0.944
0.000
0.246
5.268

KTINGSGSPHSLYWNQQT
3856
53.262
0.000
0.000
0.000
0.000
0.000

KTINGSGSPHSKPKRQQT
3857
52.881
2.832
0.000
0.000
0.000
0.000

KPRWGSGSPHSKAQNQQT
3858
51.637
0.386
0.000
0.000
375.537
0.000

KTINGSGSPHSKAFSWQT
3859
51.304
1.805
0.000
0.000
0.000
0.000

KTINGSGSPHSRFWNQQT
3860
51.225
6.955
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQCLKT
3861
49.565
1.453
0.000
0.000
0.000
0.000

KTINGSGSPHSRMRNQQT
3862
48.902
2.816
0.000
0.000
0.000
0.000

KTINGSGSPHSVKKNQQT
3863
48.475
3.908
0.000
0.000
0.000
0.000

KTINGSGSPHSWAPNQQT
3864
47.897
1.789
0.000
0.000
0.000
0.000

KTINGSGSPHSLWKNQQT
3865
45.796
4.010
0.000
0.000
0.000
0.000

KTINGSGSPHSKARWQQT
3866
45.017
2.377
0.000
0.000
0.000
0.000

KTINGSGSPHSFRPNQQT
3867
44.801
9.191
0.000
0.000
0.000
0.000

KTINGSGSPHSKKVFQQT
3868
43.747
4.480
0.000
0.000
0.000
0.000

KTINGSLSPHFWAQNQQT
4013
43.190
2.041
0.000
0.000
0.000
0.000

KTINGSGSPHSYAFNQQT
3869
43.037
1.742
0.000
0.000
0.000
0.000

KTINHRISPHSKAQNQQT
4014
42.998
1.876
0.000
0.000
0.000
0.000

KTINGSGSPHSKACSRQT
3870
42.696
2.468
0.000
0.000
0.000
0.000

KTRRPSGSPHSKAQNQQT
4015
42.374
2.384
0.000
0.000
0.000
0.000

KYSAGSGSPHSKAQNQQT
3871
41.310
1.824
0.000
0.000
0.000
0.000

KTINGAYSPHRKAQNQQT
4016
40.969
1.283
0.000
0.000
0.000
0.000

KTINGSGSPHSKRLWQQT
3872
40.932
4.801
0.000
0.000
0.000
0.000

KTINGSGSPHSCSRNQQT
3873
40.372
4.293
0.000
0.000
0.000
0.000

KTINGSGSPHSRCPNQQT
3874
39.529
4.890
0.000
0.000
0.000
0.000

KTINGSGSPHSGACNQQT
3875
39.163
3.215
0.000
0.000
0.000
4733.916

KYYTGSGSPHSKAQNQQT
3876
38.777
1.199
0.000
0.000
0.000
0.000

KTINGSGSPHSKFRQQQT
3877
38.665
3.260
0.000
0.000
0.000
0.000

KTINGSGSPHSFPFNQQT
3878
38.584
4.693
0.000
0.000
0.000
0.000

KTINGSGSPHSFFGNQQT
3879
38.088
6.101
0.000
0.000
0.000
0.000

KTINGRRSPHGKAQNQQT
4017
37.728
3.259
0.000
0.000
0.000
0.000

KTINGSGSPHSMCQNQQT
3880
37.209
1.348
0.000
0.000
0.000
0.000

KTINGSGSPHSKLFWQQT
3881
37.022
4.178
0.000
0.000
0.000
0.000

KTINGSGSPHSKTRKQQT
3882
36.010
2.858
0.000
0.000
0.000
0.000

KTINGRTSPHRKAQNQQT
4018
35.792
5.682
0.000
0.000
0.000
0.000

KTINGSGSPHSGKRNQQT
3883
35.120
5.396
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQNFKR
3884
32.291
0.964
0.000
0.000
0.000
0.000

KTINGSGSPHFYRQNQQT
3885
31.724
9.342
0.000
0.000
0.000
0.000

KTINGSRSPHAWAQNQQT
4019
31.146
6.838
0.000
0.000
0.000
0.000

KTINGSGSPHCRVQNQQT
3886
31.043
1.203
0.000
0.000
0.000
0.000

KTINGSGSPHYGIQNQQT
3887
30.908
1.076
0.000
0.000
0.000
0.000

KTINKCLSPHSKAQNQQT
4020
30.667
5.097
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQRFKT
3888
30.363
0.139
0.000
0.000
0.000
0.000

KTINGSGSPHVNCQNQQT
3889
30.010
6.122
0.000
0.000
0.000
0.000

KTINGSGSPHSKPFRQQT
3890
29.842
8.700
0.000
0.000
0.000
0.000

KTINGSGSPHSLAWNQQT
3891
29.015
4.746
0.000
0.000
0.000
0.000

KTINGSGSPHSKRSYQQT
3892
28.973
2.116
0.000
0.000
0.000
0.000

KTINGSSSPHRCAQNQQT
4021
28.887
1.829
0.000
0.000
0.000
0.000

KTINGSGSPHWSYQNQQT
3893
28.607
3.751
0.000
0.000
0.000
0.000

KTINCRTSPHSKAQNQQT
4022
28.301
1.117
0.000
0.000
0.000
0.000

KTINGSGSPHRWLQNQQT
3894
28.147
6.882
0.000
0.000
0.000
0.000

KTIFDCGSPHSKAQNQQT
4023
27.844
1.602
0.000
0.000
0.000
0.000

KTINGSGSPHPSCQNQQT
3895
27.796
2.790
0.000
0.000
0.000
0.000

KTINGSGSPHSSWLNQQT
3896
27.318
3.271
0.000
0.000
0.000
0.000

KTINSPRSPHSKAQNQQT
4024
27.240
1.554
0.000
0.000
0.000
0.000

KPRFGSGSPHSKAQNQQT
3897
27.203
0.657
0.000
0.000
0.000
0.000

KWLTGSGSPHSKAQNQQT
3898
26.975
2.388
0.364
0.000
0.000
2578.486

KTINGSGSPHSKRRAQQT
3899
26.523
5.906
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQTCRT
3900
26.472
6.369
0.000
0.000
0.000
0.000

KTINGLDSPHRSRQNQQT
4025
26.403
0.321
0.000
0.000
0.000
0.000

KTINGSGSPHSKGCTQQT
3901
26.068
0.529
0.000
0.000
0.000
0.000

KTRTRSGSPHSKAQNQQT
4026
25.852
6.894
0.000
0.000
0.000
0.000

KTINGSGSPHVPWQNQQT
3902
25.294
3.435
0.000
0.000
0.000
0.000

KTINGSGSPHSKRYTQQT
3903
25.267
9.412
0.000
0.000
0.000
0.000

KTINGSISPHCPAQNQQT
4027
24.932
0.556
0.000
0.000
0.000
0.000

KTINGSGSPHSGCQNQQT
3904
24.818
1.981
0.000
0.000
0.000
0.000

KTINGSGSPHSFTPNQQT
3905
24.227
1.036
0.000
0.000
0.000
0.000

KTINGSGSPHSTTCNQQT
3906
23.771
3.315
0.000
0.000
0.000
0.000

KTINGSGSPHSKARMYQT
3907
23.424
0.313
0.000
0.000
0.000
0.000

KTINGLVSPHRKAQNQQT
4028
23.417
2.739
0.000
0.000
0.000
0.000

KTINGSGSPHPKRQNQQT
3908
23.055
2.355
0.000
0.000
0.000
0.000

KTINGSGSPHSKCFLQQT
3909
22.987
1.434
0.000
0.000
0.000
0.000

KTINGSGSPHWVPQNQQT
3910
22.907
3.219
0.000
0.000
0.000
0.000

KTINGSGSPHSFWSNQQT
3911
22.857
1.345
0.000
0.000
0.000
0.000

KRSYGSGSPHSKAQNQQT
3912
22.474
2.841
0.000
0.000
0.000
0.000

KYVFGSGSPHSKAQNQQT
3913
22.232
2.346
0.000
0.000
0.000
0.000

KTINGSGSPHSKFKNQQT
3914
21.951
1.074
0.000
0.000
0.000
0.000

KTINGSGSPHRIKQNQQT
3915
21.720
3.064
0.000
0.000
0.000
0.000

KTINGSGSPHSKAPRRQT
3916
21.645
3.940
0.000
0.000
0.000
0.000

KTINGSGSPHSFRYNQQT
3917
21.097
4.148
0.000
0.000
0.000
0.000

KTINGSGSPHSKMICQQT
3918
21.036
0.144
0.000
0.000
0.000
0.000

KTINGSGSPHLRWQNQQT
3919
21.014
9.649
0.000
0.000
0.000
0.000

KTINGSGSPHLPTQNQQT
3920
20.704
3.127
0.000
0.000
0.000
0.000

KTINGSGSPHSKWKSQQT
3921
20.390
1.239
0.000
0.000
4.904
0.163

KTINALRSPHSKAQNQQT
4029
20.053
1.655
0.000
0.000
0.000
0.000

KTINGSGSPHSYMRNQQT
3922
20.007
2.293
0.000
0.000
0.000
0.000

KTINGSGSPHSKAARRQT
3923
19.998
6.633
0.000
0.000
0.000
0.000

KTINGSGSPHLLCQNQQT
3924
19.796
3.484
0.673
0.000
0.000
1.309

KTINGSGSPHRCCQNQQT
3925
19.084
2.213
0.000
0.000
0.000
0.000

KTINGSGSPHLCVQNQQT
3926
19.030
1.428
0.000
0.000
0.000
0.000

KTINGSGSPHSKLTRQQT
3927
19.004
2.712
0.000
0.000
0.000
0.000

KTICGRGSPHSKAQNQQT
4030
18.923
2.171
0.000
0.000
0.000
0.000

KTTRKSGSPHSKAQNQQT
4031
18.849
2.617
0.000
0.000
0.000
0.000

KTINGSGSPHSKLCTQQT
3928
18.674
1.269
0.000
0.000
0.000
0.000

KKHLGSGSPHSKAQNQQT
3929
18.521
0.658
0.000
0.000
0.000
0.000

KTINGSGSPHSKIRGQQT
3930
18.150
1.584
0.000
0.000
0.000
0.000

KTMQRSGSPHSKAQNQQT
4032
18.020
3.159
0.000
0.000
0.000
0.000

KTINGSGSPHSYLVNQQT
3931
17.766
1.267
0.000
0.000
0.000
0.000

KTINGSGSPHQGCQNQQT
3932
17.676
1.037
0.000
0.000
0.000
0.000

KTINGSGSPHMAFQNQQT
3933
17.644
0.542
0.000
0.000
0.000
0.000

KTINGSGSPHSKACQFQT
3934
17.640
8.562
0.000
0.000
0.000
9.605

KTINGSGSPHSKWGLQQT
3935
17.543
2.639
0.000
0.000
0.000
0.000

KTINGSGSPHSKILRQQT
3936
17.419
2.546
0.000
0.000
0.000
0.000

KTINGSGSPHSFQINQQT
3937
17.418
0.269
0.308
0.000
1.568
0.000

KTINGSGSPHSKACISQT
3938
17.371
0.240
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQTHRT
3939
17.290
2.917
0.000
0.000
0.000
0.000

KTINGSGSPHSKALRCQT
3940
17.283
1.892
0.000
0.000
0.000
0.000

KTINGSGSPHSKAFYIQT
3941
17.172
0.239
0.000
0.000
0.000
0.000

KTINGSGSPHSKAHARQT
3942
17.075
1.800
0.000
0.000
0.000
0.000

KTINGSGSPHSLCLNQQT
3943
17.028
1.790
0.000
0.000
0.000
0.000

KTINGSGSPHSKAFVRQT
3944
16.935
1.985
0.000
0.000
0.000
0.000

KPPLGSGSPHSKAQNQQT
3945
16.897
0.805
0.000
0.000
0.000
0.000

KTINGSGSPHRPWQNQQT
3946
16.869
4.936
0.000
0.000
0.000
0.000

KPARGSGSPHSKAQNQQT
3947
16.793
1.391
0.000
0.000
0.000
0.000

KTINGSGSPHRPRQNQQT
3948
16.784
5.206
0.000
0.000
0.000
0.000

KTINGSGSPHSCPQNQQT
3949
16.701
1.776
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQFILT
3950
16.650
4.208
0.000
0.000
0.000
0.000

KTINGSSSPHWMAQNQQT
4033
16.635
2.390
0.000
0.000
0.000
0.000

KTRKRSGSPHSKAQNQQT
4034
16.603
2.075
0.000
0.000
0.000
0.000

KTINGSGSPHSVRYNQQT
3951
16.390
1.413
0.000
0.000
0.000
0.000

KSRRGSGSPHSKAQNQQT
3952
16.131
1.446
0.000
0.000
0.000
0.000

KTINGSGSPHSVRCNQQT
3953
15.860
3.912
0.000
0.000
0.000
0.000

KFFHGSGSPHSKAQNQQT
3954
15.412
0.897
0.000
0.000
0.000
0.000

KTINGSGSPHSKMPCQQT
3955
15.343
1.063
0.000
0.000
0.000
0.000

KTINGSGSPHSKKTSQQT
3956
15.244
1.344
0.000
0.000
0.000
0.000

KRYNGSGSPHSKAQNQQT
3957
15.160
0.806
0.000
0.000
0.000
0.000

KTINFTRSPHSKAQNQQT
4035
14.908
3.751
0.000
0.000
0.000
0.000

KTINGSGSPHSLPYNQQT
3958
14.792
2.048
0.000
0.000
0.000
0.000

KTINGSGSPHVYHQNQQT
3959
14.770
1.733
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQSRKT
3960
14.589
2.715
0.000
0.000
0.000
0.000

KTINGSGSPHSYTRNQQT
3961
14.535
1.986
0.000
0.000
0.000
0.000

KTINNLRSPHSKAQNQQT
4036
14.514
1.354
0.000
0.000
0.000
0.000

KTINGRPSPHGKAQNQQT
4037
14.442
0.705
0.000
0.000
0.000
0.000

KTINWSRSPHSKAQNQQT
4038
14.399
5.624
0.000
0.000
0.000
0.000

KTINGSGSPHLVYQNQQT
3962
14.196
1.045
0.000
0.000
0.000
0.000

KTINGTRSPHKKAQNQQT
4039
14.173
1.152
0.700
0.225
0.052
4.082

KTINGSGSPHSKALRWQT
3963
14.118
5.252
0.000
0.000
0.000
0.000

KTINGSGSPHYRYQNQQT
3964
14.107
1.027
0.000
0.000
0.000
0.000

KTINGSGSPHSWLKNQQT
3965
13.995
0.603
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQMQIT
3966
13.990
0.371
0.000
0.000
0.000
0.000

KTINGSVSPHCTAQNQQT
4040
13.502
2.955
0.000
0.000
0.000
0.000

KTINGSGSPHCPAQNQQT
3967
13.359
1.409
0.000
0.000
0.000
0.000

KTINGSGSPHSMCTNQQT
3968
13.114
0.392
0.000
0.000
0.000
0.000

KTINGSGSPHSPPDNQQT
3969
12.973
0.033
0.000
0.000
0.000
0.000

KTINGSGSPHSKRNYQQT
3970
12.781
5.528
0.000
0.000
0.000
0.000

KTTRCSGSPHSKAQNQQT
4041
12.639
8.168
0.000
0.000
0.000
0.000

KTKLCSGSPHSKAQNQQT
4042
12.570
2.139
0.000
0.000
0.000
0.000

KTINLGCSPHSKAQNQQT
4043
12.564
0.654
0.000
0.000
0.000
0.000

KTINGSGSPHRWTQNQQT
3971
12.490
0.844
0.000
0.000
0.000
0.000

KTISGHDSPHISGQYQQT
4044
12.395
0.420
0.000
0.000
0.074
1214.588

KTINGSGSPHSKACRLQT
3972
12.297
6.537
0.000
0.000
0.000
0.000

KTINGSGSPHPRKQNQQT
3973
12.249
3.248
0.000
0.000
0.000
0.000

KTINGSGSPHSKCSVQQT
3974
12.246
1.465
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQYVRT
3975
12.239
3.275
0.000
0.000
0.000
0.000

KTINGSGSPHSKARISQT
3976
12.142
1.565
0.000
0.000
0.000
0.000

KTINGRRSPHMKAQNQQT
4045
12.136
3.510
0.000
0.000
0.000
0.000

KTINGPWSPHRKAQNQQT
4046
12.103
0.434
0.000
0.000
0.000
0.000

KTINGSGSPHPFVQNQQT
3977
12.091
1.286
0.000
0.000
0.000
0.000

KTINGSGSPHSKLPKQQT
3978
11.856
0.274
0.000
0.000
0.000
0.000

KTINSCFSPHSKAQNQQT
4047
11.847
1.016
0.000
0.000
0.000
0.000

KTINGSGSPHSKSEQQQT
3979
11.785
1.769
0.000
0.000
0.000
0.000

KTINGSGSPHWVAQNQQT
3980
11.703
3.634
0.000
0.000
0.000
0.000

KTINGSGSPHSLYQNQQT
3981
11.590
1.503
0.000
0.000
0.000
0.000

KTINGSGSPHSKVRMQQT
3982
11.572
1.835
0.000
0.000
0.000
0.000

KTINYTRSPHSKAQNQQT
3983
11.514
0.431
0.000
0.000
0.000
0.000

KTIKRYGSPHSKAQNQQT
4048
11.461
2.022
0.000
0.000
0.000
0.000

KTINGSGSPHCALQNQQT
4693
11.404
3.867
0.000
0.000
0.000
0.000

KTINGSGSPHSSCTNQQT
3984
11.382
3.363
0.000
0.000
0.000
0.000

KTINGSGSPHSKNSRQQT
3985
11.280
1.093
0.000
0.000
0.000
0.000

KTINGSGSPHSKRKRQQT
3986
11.215
3.027
0.000
0.000
0.000
0.000

KTINGSGSPHLCTQNQQT
3987
11.176
2.489
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQSAKT
3988
11.162
4.200
0.000
0.000
0.000
0.000

KTINGSGSPHSTCLNQQT
3989
11.132
4.762
0.000
0.000
0.000
0.000

KTINGSGSPHSYARNQQT
3990
11.131
0.996
0.000
0.000
0.000
0.000

KTINGSGSPHSKQRPQQT
3991
11.130
2.347
0.000
0.000
0.000
0.000

KTINGSGSPHSKRVVQQT
3992
11.094
1.639
0.000
0.000
0.000
0.000

KRFSGSGSPHSKAQNQQT
3993
11.024
1.358
0.000
0.000
0.000
0.000

KTINGSGSPHKSGQNPQT
3994
11.014
11.790
0.000
0.000
0.000
0.000

KTINRYSSPHSKAQNQQT
4049
10.926
1.544
0.000
0.000
0.000
0.000

KTTGRSGSPHSKAQNQQT
4050
10.863
0.126
0.000
0.000
0.000
0.000

KTINGSGSPHSKALRHQT
3995
10.774
4.532
0.000
0.000
0.000
0.000

KTINGSGSPHSYYSNQQT
3996
10.680
2.856
0.000
0.000
0.000
0.000

KTINGSGSPHSLTCNQQT
3997
10.658
2.214
0.490
0.000
0.163
1.398

KTINGSGSPHSCQSNQQT
3998
10.631
1.468
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQSIKT
3999
10.544
1.355
0.000
0.000
0.000
0.000

KYSMGSGSPHSKAQNQQT
4000
10.478
1.587
0.000
0.000
0.000
0.000

KTINGSGSPHSKAKGWQT
4001
10.450
1.827
0.000
0.000
0.000
0.000

KTIVGSGSPHSKPQNQQT
4002
10.381
0.894
0.000
0.000
0.000
0.000

KTINGSGSPHFPFQNQQT
4003
10.322
3.715
0.000
0.000
0.000
0.000

KPFLGSGSPHSKAQNQQT
4004
10.318
1.328
0.000
0.000
0.000
0.000

KTINGSGSPHSKCTSQQT
4005
10.311
5.821
0.493
0.232
1.413
2.353

KTINRQFSPHSKAQNQQT
4051
10.275
4.480
0.000
0.000
0.000
0.000

KTINGSGSPHSVFENQQT
4006
10.218
0.224
0.000
0.000
0.000
0.000

KTINGSGSPHSKAKKVQT
4007
10.102
3.974
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQRCST
4008
10.084
0.762
0.000
0.000
0.000
0.000

KTINGSGSPHSKAQFCLT
4009
10.065
3.371
0.000
0.000
0.000
0.000

KTINGSGSPHGRYQNQQT
4010
10.028
0.778
0.000
0.000
0.000
0.000

Table 18 provides the peptide sequences of 43 matured capsid variants having a raw virus count greater than 10, a CV of less than 1 for the heart samples isolated, and that also demonstrated a 4-fold or greater fold-increase in expression in the heart relative to the AAV9 control. A number of the matured variants shown in Table 18 also demonstrated increased expression in other tissues isolated from the NHPs, including the brain, muscle, and/or liver, and are therefore pan-tropic.

TABLE 18

NGS fold-enrichment of TTM-001 and TTM-002 matured AAV capsid variants in the

heart of NHPs

Fold Enrichment relative to AAV9

SEQ

Liver
Liver

ID
Heart
Brain
DRG
Muscle
RNA
DNA
Brain

Sequence
NO:
(NHP)
(NHP)
(NHP)
(NHP)
(NHP)
(NHP)
(Mouse)

KTITGHDSPHSKAQNQQT
4052
34.375
230.437
4.338
1.378
19.165
5.672
0.000

KTINGSGSPHKSGQYQQT
4053
33.208
851.414
8.704
17.754
17.342
9.915
12.911

KTINGSGSPHKSGQDQQT
4054
31.166
218.057
34.358
33.372
27.081
8.836
24.849

KTINGSGSPHKSGQIQQT
4055
27.293
201.467
48.033
12.706
17.874
13.192
10.912

KTINGSGSPHKSGRNQQT
4056
27.283
313.826
8.723
36.593
15.252
12.352
21.595

KTINGSGSPHKSGKNQQT
4057
25.992
230.621
6.343
97.671
15.369
7.226
31.282

KTIYGHDSPHSKAQNQQT
4058
25.673
269.879
3.694
8.391
11.895
6.197
0.000

KTINGSGSPHKSGQNQQS
4059
24.783
244.030
16.675
26.058
18.059
9.809
29.751

KTINGSGSPHKSGQNLQT
4060
24.464
392.519
15.629
0.371
29.977
18.332
30.446

KAINGHDSPHSKAQNQQT
4061
22.460
640.466
7.358
9.986
9.358
8.490
0.000

KTVNGHDSPHSKAQNQQT
4062
21.066
614.034
3.392
30.908
21.560
11.933
121.235

KTIKGHDSPHSKAQNQQT
4063
20.803
213.564
24.646
12.361
15.379
6.551
13.319

KSINGHDSPHSKAQNQQT
4064
20.698
246.819
7.592
28.235
11.773
6.888
280.630

KTINGSGSPHKSGQTQQT
4065
19.925
466.459
55.454
15.485
15.473
6.446
15.179

KTINGSGSPHKSGHNQQT
4066
19.548
287.922
12.159
20.851
17.821
10.084
21.011

KTFNGHDSPHSKAQNQQT
4067
19.301
239.922
9.109
17.215
12.193
6.413
30.747

KTINGSGSPHKSGLNQQT
4068
19.136
319.093
3.083
4.096
14.009
7.446
9.340

KTINGHDSPHSKALNQQT
4069
18.542
605.641
13.375
1.902
12.621
7.054
51.283

KTINGSGSPHKSGQNQLT
4070
18.454
317.452
33.967
28.952
18.533
8.992
36.272

KTLNGHDSPHSKAQNQQT
4071
18.236
195.734
19.341
9.266
25.732
13.333
0.000

KTINGSGSPHKSGQNQHT
4072
14.269
313.837
7.125
39.273
29.714
7.797
25.119

KTIDGHDSPHSKAQNQQT
4073
13.836
242.100
1.731
12.555
17.223
7.439
0.000

KTNNGHDSPHSKAQNQQT
4074
12.872
134.488
0.504
3.877
17.044
5.982
22.358

KTINGSGSPHKSGQKQQT
4075
12.357
323.373
10.936
1172.3
12.604
7.970
48.699

KTINGSGSPHKSGQNRQT
4076
11.563
145.363
36.865
3.855
11.403
7.667
16.860

KTINGSGSPHKSGQNQQN
4077
11.507
156.385
582.38
8.559
9.273
7.668
18.138

KTINGSGSPHKSGQNQQA
4078
11.313
135.164
12.425
12.699
9.714
6.077
17.265

KTINGHDSPHSKAHNQQT
4079
10.024
236.106
19.495
5.258
2.406
3.316
45.691

KTINGHDSPHSKAQNQQT
4080
8.954
186.839
9.457
5.507
5.929
3.651
31.453

KTINGSGSPHKSGQNQQP
4081
8.744
261.947
43.435
10.217
6.468
4.265
19.828

KTINGHDSPHCKAQNQQT
4082
8.417
15.165
0.887
2.368
3.328
0.771
148.172

KTINGHDSPHSKAQNQQS
4083
5.678
603.027
7.280
0.670
4.301
4.307
65.271

KTINGSGSPHKSGQNQQT
4084
5.586
115.994
28.397
4.326
5.307
3.569
24.908

KTINGHDSPDKSGQNQQT
4085
5.569
30.854
4.934
1.112
0.671
0.781
14.499

KPINGHDSPHKSGQNHQS
4086
5.203
36.266
0.000
0.258
4.478
0.521
28.786

KTSNGSGSPHKSGQNQQT
4087
4.746
197.282
4.177
4.466
3.972
7.425
75.623

KTVNGSGSPHKSGQNQQT
4088
4.610
200.076
2.739
2.873
2.725
3.478
43.548

KTINGHDSTHKSGHNQQT
4089
4.369
27.630
2.883
1.302
0.421
0.176
12.973

KTINGHDSPHSKAQNQQN
4090
4.271
319.610
1.163
5.173
3.406
4.995
50.220

KTIYGSGSPHKSGQNQQT
4091
4.140
110.329
2.603
2.545
4.488
4.110
29.293

KTINGLDSQHKSGQNQQT
4092
4.055
12.958
3.240
3.205
0.645
0.296
5.608

Table 19 provides the peptide sequences of 14 matured capsid variants having a raw virus count greater than 10, a CV of less than 1 for the muscle samples isolated (e.g., quadriceps), and that also demonstrated a 4-fold or greater fold-increase in expression in the muscle relative to the AAV9 control. A number of the matured variants shown in Table 19 also demonstrated increased expression in other tissues isolated from the NHPs, including the brain, heart, and/or liver, and are therefore pan-tropic.

TABLE 19

NGS fold-enrichment of TTM-001 and TTM-002 matured AAV capsid variants in the

muscle (e.g., quadriceps) of NHPs

Fold Enrichment relative to AAV9

SEQ

Liver
Liver

ID
Muscle
Brain
DRG
Heart
RNA
DNA
Brain

Sequence
NO:
(NHP)
(NHP)
(NHP)
(NHP)
(NHP)
(NHP)
(Mouse)

KTINGSGSPHKSGRNQQT
4056
36.593
313.826
8.723
27.283
15.252
12.352
21.595

KTIIGHDSPHSKAQNQQT
4095
27.271
341.528
5.423
26.154
18.305
6.293
0.000

KTIYGHDSPHSKAQNQQT
4058
8.391
269.879
3.694
25.673
11.895
6.197
0.000

KTINGSGSPHKSGONQQS
4059
26.058
244.030
16.675
24.783
18.059
9.809
29.751

KTVNGHDSPHSKAQNQQT
4062
30.908
614.034
3.392
21.066
21.560
11.933
121.235

KTIKGHDSPHSKAQNQQT
4063
12.361
213.564
24.646
20.803
15.379
6.551
13.319

KSINGHDSPHSKAQNQQT
4064
28.235
246.819
7.592
20.698
11.773
6.888
280.630

KTINGSGSPHKSGHNQQT
4066
20.851
287.922
12.159
19.548
17.821
10.084
21.011

KTFNGHDSPHSKAQNQQT
4067
17.215
239.922
9.109
19.301
12.193
6.413
30.747

KTSNGHDSPHSKAQNQQT
4096
18.580
507.189
7.777
17.770
21.537
8.789
70.219

KTINGHDSPHSKAQNQQT
4080
5.507
186.839
9.457
8.954
5.929
3.651
31.453

KTINGSGSPHKSGONOQT
4084
4.326
115.994
28.397
5.586
5.307
3.569
24.908

KTINGHDSPHSKAQNQON
4090
5.173
319.610
1.163
4.271
3.406
4.995
50.220

KTINGSGSPHSKAQNRRR
4097
4.237
8.348
0.291
0.636
1.597
5.396
158.853

Additional variants were identified following generation and screening in NHPs that had the following properties. TTM-001 and TTM-002 capsid variants comprising the amino acid sequence of SEQ ID NOs: 4253, 4281, 4290-4295, 4304, 4305, 4320, 4328-4335, 4337-4340, 4353, 4355, 4369, 4387, 4421, 4424-4428, 4430, 4432, 4433, 4435, 4436-4449, 4452, 4455, 4476, 4483, or 4484 had a raw virus count 10 or greater, a CV of less than 1 for the brain samples isolated from the NHPs, demonstrated a 50-fold or greater increase in expression in the brain of mice and NHPs relative to AAV9, and demonstrated 2-fold or less expression in the liver and DRG of NHPs relative to AAV9. TTM-001 and TTM-002 capsid variants comprising the amino acid sequence of SEQ ID NOs: 4098-4105, 4254-4280, 4282-4289, 4296-4303, 4306-4327, 4336, 4341-4352, 4354, 4356-4420, 4422, 4423, 4425, 4429, 4431, 4434, 4444, 4450, 4451, 4453, 4454, 4456-4475, 4477-4482, or 4485 had a CV of less than 1 in across the brain samples isolated from the NHPs and demonstrated a 100-fold or greater increase in expression in the brain of NHPs relative to AAV9. TTM-001 and TTM-002 capsid variants comprising the amino acid sequence of SEQ ID NOs: 4102 and 4106-4252 had normalized virus counts of greater than or equal to 0.01, a CV of less than 1 across the liver RNA samples isolated from the NHPs, and demonstrated a 20-fold or greater increase in expression in the liver of NHPs relative to AAV9. TM-001 and TTM-002 capsid variants comprising the amino acid sequence of SEQ ID NO: 4105 had a raw virus count 9.9 or greater, a CV of less than 1 across the muscle samples isolated from the NHPs, and 5-fold or greater increase in expression in the muscle of the NHPs relative to AAV9. TM-001 and TTM-002 capsid variants comprising the amino acid sequence of SEQ ID NO: 4105 also had a raw virus count 9.9 or greater, a CV of less than 1 across the samples isolated from the heart of the NHPs, and 5-fold or greater increase in expression in the heart of the NHPs relative to AAV9.

These data demonstrate that following two maturation approaches, matured TTM-001 and TTM-002 capsid variants (AAV9 capsid variants) with loop IV modifications were generated with significantly enhanced CNS tropism over wild-type AAV9 controls in both NHPs and mice, while also exhibiting de-targeting in peripheral tissues (e.g., the liver and DRG). These resulting matured variants therefore demonstrated cross-species CNS tropism in both NHPs and mice. Matured TTM-001 and TTM-002 capsid variants with liver-specific tropism were also generated with at least 10 times the expression compared to wild-type AAV9 in the liver of NHPs. Several matured variants were also generated with increased expression in the heart and skeletal muscle (e.g., quadriceps) relative to wild-type AAV9 in NHPs.

Example 5. Evaluation of TTM-001 and TTM-002 AAV Capsid Variants in Diverse Primate Species

This Example evaluates the tropism and cross-species compatibility of the TTM-001 (SEQ ID NO: 981 (amino acid) and 983 (DNA), comprising SEQ ID NO: 941) and TTM-002 (SEQ ID NO: 982 (amino acid) and 984 (DNA), comprising SEQ ID NO: 2) capsid variants in two diverse primate species, marmosets (Callithrix jacchus) and African green monkeys (Chlorocebus sabaeus), as compared to their tropism in cynomolgus macaques (Macacafascicularis) provided in Example 1. The cross-species compatibility and tropism of an AAV9 capsid variant comprising the amino acid sequence of SPHKYG (SEQ ID NO: 966) was also investigated in this example. The amino acid and DNA sequences of TTM-001 and TTM-002 are provided, e.g., in Tables 4 and 5, respectively.

To investigate tropism in African green monkeys, AAV particles comprising the TTM-001 capsid variant, the TTM-002 capsid variant, an AAV9 capsid variant comprising SEQ ID NO: 966, or an AAV9 control under the control of a synapsin promoter, were intravenously injected into NHPs (n=2, 3-12 years of age) at a dose of 2E13 vg/kg. After 14-days in life, the brains and tissues (liver, DRG, quadriceps, and heart) of the NHPs were collected and RNA was extracted. Following RNA recovery and RT-PCR amplification, a systematic NGS enrichment analysis was performed to calculate the fold enrichment ratio relative to the AAV9 wild-type control.

To investigate tropism in marmoset monkeys, AAV particles comprising the TTM-001 capsid variant, the TTM-002 capsid variant, an AAV9 capsid variant comprising SEQ ID NO: 966, or an AAV9 control, were intravenously injected into NHPs (n=2, >10 months of age) at a dose of 2E13 vg/kg (8.75E12 vg/mL). After 28-days in life, the brains and tissues (liver quadriceps, and heart) of the NHPs were collected and RNA was extracted. Following RNA recovery and RT-PCR amplification, a systematic NGS enrichment analysis was performed to calculate the fold enrichment ratio relative to the AAV9 wild-type control.

As provided in Table 20 (African green monkeys) and Table 21 (marmosets), both the TTM-001 and TTM-002 capsid variants demonstrated increased CNS tropism in diverse primate species. The TTM-001 capsid variant demonstrated a 73.6-fold increase in expression relative to AAV9 in the brain of cynomolgus macaques (Table 9, Example 1), a 43.5-fold increase in expression relative to AAV9 in the brain of African green monkeys, and a 703.3-fold increase in expression relative to AAV9 in the brain of marmosets. The TTM-002 capsid variant demonstrated a 62.6-fold increase in expression relative to AAV9 in the brain of cynomolgus macaques (Table 9), a 13.8-fold increase in expression relative to AAV9 in the brain of African green monkeys, and a 366.6-fold increase in expression relative to AAV9 in the brain of marmosets. Both TTM-001 and TTM-002 led to a significant increase in expression relative to AAV9 in the heart of both African green monkeys and marmosets (Table 20 and Table 21). The AAV9 capsid variant comprising SEQ ID NO: 966 also demonstrated in increase in expression relative to AAV9 in the brain and heart of both African green monkeys and marmosets. Furthermore, TTM-001, TTM-002, and the AAV9 capsid variant comprising SEQ ID NO: 966, also all led to increased expression in the brain of both BALB/c and C57B1/6 mice (Table 11, Example 1), demonstrating an average fold change in expression relative to AAV9 across both species of mice of 63.1, 66.8, and 126.97, respectively.

TABLE 20

NGS-fold enrichment of TTM-001 (comprises SEQ ID NO: 941),

TTM-002 (comprises SEQ ID NO: 2), and an AAV9 capsid

variant comprising SEQ ID NO: 966 in African green monkeys

SEQ ID
Fold Enrichment relative to AAV9

Sequence
NO:
Brain
DRG
Heart
Liver DNA
Liver RNA
Muscle

SPHSKA
941
43.525
1.010
184.789
0.242
1.547
1.715

HDSPHK
2
13.779
0.678
35.991
0.084
0.087
0.144

SPHKYG
966
9.805
0.071
44.865
0.085
0.136
0.234

TABLE 21

NGS-fold enrichment of TTM-001 (comprises SEQ ID NO: 941),

TTM-002 (comprises SEQ ID NO: 2), and an AAV9 capsid

variant comprising SEQ ID NO: 966 in marmosets

SEQ ID
Fold Enrichment relative to AAV9

Sequence
NO:
Brain
Heart
Liver DNA
Liver RNA
Muscle

SPHSKA
941
703.610
48.979
0.268
0.779
0.425

HDSPHK
2
366.625
18.572
0.075
0.276
0.229

SPHKYG
966
150.209
17.232
0.045
0.014
0.146

Taken together, these data demonstrate that the AAV9 capsid variants of TTM-001 and TTM-002 demonstrated increased CNS tropism relative to the AAV9 control in the CNS across three diverse primate species and two species of mice, providing evidence of strong cross-species capacity. The AAV9 capsid variant comprising the amino acid sequence of SEQ ID NO: 966 also demonstrated strong CNS expression relative to the AAV9 control in two species of NHPs and two species of mice, also showing strong cross-species capacity.

Example 6. Advanced Maturation of TTM-002 Capsid Variant in Mice

This Example describes additional maturation of the TTM-002 (SEQ ID NO: 982 (amino acid) and 984 (DNA), comprising SEQ ID NO: 2) capsid variant in mice. In order to mature the TTM-002 capsid variant, sets of three contiguous amino acids were randomized across the mutagenesis region in TTM-002 sequence, which spanned from position 450 to position 466, numbered according to SEQ ID NO: 982. Unlike the maturation performed in in Example 3, where the SPH motif that was observed in the AAV capsid variants that demonstrated the greatest fold-enrichment in the NHP brain relative wild-type AAV9 was not disrupted, in the maturation approach used in this Example, the SPH motif was not held constant to further explore the role of this motif in the capsid variant. The matured TTM-002 capsid variants that resulted from the maturation approach were pooled together for subsequent testing and characterization in mice.

The library of matured AAV capsid variants generated from the TTM-002 matured AAV capsid variant were intravenously injected into the tail vein of three CD-1 Outbred mice (Charles River; 6-8 weeks of age) at a dose of 1.0×10¹²VG/dose. After about 28 days in life, the brains of the mice were isolated, and RNA was extracted. Following RNA recovery and RT-PCR amplification, a systematic NGS enrichment analysis was performed to calculate the fold enrichment ratio relative to the corresponding TTM-002 non-matured control, and the peptides comprised within the variants were identified. Variants were filtered by those with a raw virus count in the sample above 10 and a coefficient of variance (CV) that was greater than 1 (identifies the peptides/variants reliably detected in the majority of the samples isolated from the three mice).

Following the advanced maturation screen and filtering of the variants, 1302 variants demonstrated an increase in expression relative to the non-matured TTM-002 capsid variant in the brain of the outbred mice. Of the 1302 variants with improved tropism relative to the non-matured TTM-002, 1283 comprised the SPH motif in the same position as the non-matured TTM-002 capsid variant (e.g., immediately subsequent to position 455, relative to a reference sequence numbered according to the amino acid sequence of SEQ ID NO: 138 or 982). Mutations in the region of the SPH motif present in the non-matured TTM-002 capsid variant only consistently appear in those variants with a fold change of 0.2 or 0.1 or lower relative to the non-matured TTM-002 control in the brain of the mice. This indicates that the SPH motif may be important to the increased brain tropism that observed for the TTM-002 capsid variant. In instances when the SPH motif was disrupted, the fold change of the matured variants of TTM-002 decreased considerably in relation to the non-matured TTM-002 variant which comprised the SPH motif.

Example 7. Tropism of TTM-002 AAV Capsid Variant

This Example further investigates the tropism and CNS cells transduced by the TTM-002 capsid variant (SEQ ID NO: 982 (amino acid) and 984 (DNA), comprising SEQ ID NO: 2), as outlined in Table 3 above. The amino acid and DNA sequences of TTM-002 are provided, e.g., in Tables 4 and 5, respectively.

AAV particles were generated with the TTM-002 capsid variant encapsulating a GFP transgene (AAV_TTM-002.GFP) or a payload driven by a heterologous CBA constitutive promoter (AAV_TTM-002.Payload).

Two tandem single cell RNA sequencing runs (scRNA-Seq) of mouse cells derived from the midbrain area were performed. In the first run, cells were pooled from two mice at day 28 post treatment with AAV_TTM-002.Payload particles. In the second run we treated with AAV_TTM-002.GFP particles, in the same manner but without xenografts. Orthotopic xenografts of MDA-MB-361-Luc #1 high passage cells grown as tumorspheres (in tumorsphere media; Sigma #C-28070) were injected (250,000 cells/2 μL/mouse) intracranially into 2-month old female SCID CB17 (Mutation: Icr-Prkdcscid/IcrIcoCrl) congenic immunodeficient mice (Charles River Laboratories). The injections were 2.5 mm (lateral), −1 mm (posterior) with respect to bregma, lowered −3 mm ventral and raised+0.5 mm dorsal to a final −2.5 mm ventral position. Two days later, dilutions of the AAV_TTM-002.Payload particles (run 1), or in the case without xenografts, dilutions of AAV_TTM-002.GFP particles (run 2) were prepared. IV injections of 100 μL (2.5e11 VG/animal) of the AAV_TTM-002.payload particles or AAV_TTM-002.GFP particles were administered through the tail veins of mice (n=5 mice per groups). At 7 days post-injection, mice from run 1 were imaged in an AmiHTX (Spectral Imager) for bioluminescence of the human tumor cells due to expression of luciferase in response to intraperitoneal luciferin injections.

At 28 days post-injection with the AAV_TTM-002.payload particles or AAV_TTM-002.GFP particles, two mice from each run were necropsied, brain samples were isolated, and the midbrain was dissected and isolated. The midbrain samples were then exposed to a cold protease inhibitor (Creative Biomart #NATE-0633) and were dissociated at 6 degrees centigrade. For the samples collected from the mice of run 1 (AAV_TTM-002.Payload particles), myelin depletion was performed (Miltenyi, #130-096-731), cells were filtered through a 40 μM mesh to filter out neurons) and loaded on a 10× chromium G chip. scRNA-Seq was performed (10× Genomics) and samples were sequenced on a NextGen500 Sequencing machine (Illumina). For the samples collected from run 2 (AAV_TTM-002.GFP particles and no xenografts), the cells were not myelin depleted or filtered through 40 μM mesh to include neurons. The cells isolated after run 2 were FACS sorted for GFP+/7AAD− (live GFP+ cells). The resultant cells were loaded on a 10× chromium G chip and the scRNA-Seq was run and processed (10× Genomics).

For run 1, the scRNA-Seq data was filtered to include cells with only greater than 1000 genes per cell and less than 5000, and less than 20 percent mitochondrial gene expression. For run 2, the scRNA-Seq data was filtered to include cells with only greater than 200 genes per cell and less than 5000, and less than 20 percent mitochondrial gene expression. The data were normalized, scaled, and integrated into one combined dataset. Clusters were generated with a resolution of 0.3 and each cluster identity was determined using a panel of cell type specific genes (e.g., as described in Brown et al., 2021. “Deep Parallel Characterization of AAV Tropism and AAV-Mediated Transcriptional Changes via Single-Cell RNA Sequencing”. Front. Immunol. 12:730825; the contents of which are hereby incorporated by reference in its entirety). The percentage of GFP sorted cells per cluster was calculated as was the percentage of payload expressing genes per cluster as parallel measures of TTM-002 transduction.

For payload expressing cells, endothelial cells had the highest proportion of payload positive cells, followed by astrocytes (Table 22). For GFP+ sorted cells, endothelial cells had the highest proportion of GFP positive cells, and astrocytes were the third highest cell type when sorting by proportion of cells expressing GFP (Table 22). These data indicate TTM-002 transduction exhibits an endothelial and astrocytic tropism. Furthermore, the astrocytic cluster had the second highest level of expression of Olig2 (oligodendrocytes demonstrated the greatest Olig2 expression). IHC staining was performed on brain samples isolated from AAV_TTM-002.GFP infected mice and demonstrated that GFP co-localized with some but not all Olig2+ cells. No co-staining was observed with mylein basic protein (MBP), a marker of oligodendrocytes. Co-staining with GFP was also not observed in NeuN positive cells (neurons), GFAP positive cells (astrocytes), and Iba1 positive cells (microglia). GFP staining was observed throughout the sagittal section of the mouse brain, which was demonstrative of increased staining in the midbrain. The GFP expressing cells observed did not have a bipolar morphology like oligodendrocyte progenitor (OPC) cells and therefore, together with the scRNA-Seq data, these results indicated that at day 28 post AAV treatment, Olig2+ astrocytes in the midbrain are being transduced by AAV particles comprising a TTM-002 capsid, in a cell type specific tropism.

TABLE 22

Quantification of payload positive cells and GFP positive cells

% Payload

% GFP

Cluster Identity
Cells/Cluster
Cluster Identity
Cells/Cluster

Endothelial-2
6.58
Endothelial-2
6.58

Astrocyte
4.50
Endothelial-1
3.45

Pericytes
4.23
Vascular and
2.38

leptomeningeal

Cells (VLM)

Mature Oligos
3.85
Astrocyte
2.37

Endothelial-1
3.09
Vascular smooth
1.03

muscle cells (VSC)

Committed Oligos
1.90
Pericytes
0.77

Vascular smooth
1.72
Microglia
0.00

muscle cells (VSC)

Microglia
0.40
Committed Oligos
0.00

Macrophages
0.00
Macrophages
0.00

Vascular and
0.00
Oligodendrocytes
0.00

leptomeningeal

cells (VLM)

Oligodendrocytes
0.00
Committed Oligos-2
0.00

Committed Oligos-2
0.00
Mature Oligos
0.00

Example 8. Individual Capsid Characterization of TTM-002 in NHPs

This example describes the transduction level, tropism, ability to cross the blood brain barrier, and overall spatial distribution in the central nervous system (CNS) and peripheral tissues of the AAV capsid variant TTM-002 (SEQ ID NO: 982 (amino acid) and 984 (DNA), comprising SEQ ID NO: 2), relative to AAV9 following intravenous administration in African green monkeys (Chlorocebus sabaeus).

AAV particles were generated with the TTM-002 capsid variant or the AAV9 capsid control which comprised a self-complementary viral genome encoding an histone H2b protein with an HA tag driven by a ubiquitous CBA promoter. The AAV particles comprising the TTM-002 capsid variant or the AAV9 capsid control were administered to the NHPs (n=2) intravenously at a dose of 1e12 VG/kg or 1e13 VG/kg. The in-life period was 28 days and then various CNS and peripheral tissues were collected for measuring transgene mRNA (expression) by RT-qPCR and viral DNA (biodistribution) by ddPCR.

As shown in Table 23, the TTM-002 capsid variant resulted in increased brain biodistribution in all brain regions investigated as compared to AAV9 at both doses tested. The TTM-002 capsid variant also led to increased transgene expression in the brain relative to AAV9 at both doses tested (Table 24). In the spinal cord, the TTM-002 capsid variant distributed to the cervical spinal cord and the spinal cord ventral horn at a higher level relative to AAV9 (Table 23) and it mediated higher transgene expression than AAV9 in both the full spinal cord and the ventral horn (Table 24). The TTM-002 capsid variant exhibited lower biodistribution (Table 23) and transgene expression (Table 24) in the DRG relative to AAV9, indicating that TTM-002 capsid variant was detargeted in the DRG relative to AAV9. Similar expression and distribution were observed by immunohistochemistry performed on these CNS tissues.

Distribution and transgene expression was also measured in the peripheral tissues of the liver, heart, and quadriceps. In the liver, TTM-002 capsid variant exhibited lower biodistribution (Table 23) and transgene expression (Table 24) relative to AAV9, indicating that TTM-002 capsid variant was detargeted in the liver relative to AAV9. In the heart, the TTM-002 capsid variant exhibited comparable levels of biodistribution relative to AAV9 (Table 23), but increased transgene expression relative to AAV9 (Table 24). In the quadriceps, TTM-002 capsid variant exhibited lower biodistribution (Table 23) and lower transgene expression (Table 24), relative to AAV9. Similar expression and distribution were observed by immunohistochemistry performed on these peripheral tissues.

TABLE 23

Quantification of viral genome copies per diploid genome (biodistribution) by ddPCR

following intravenous administration of AAV particles comprising a TTM-002 capsid