Compositions and methods for the treatment of disorders related to glucosylceramidase beta 1 deficiency

Abstract

The present disclosure relates to compositions and methods for delivering, inter alia, altering, e.g., enhancing, the content of GBA1 protein via the use of adeno-associated virus (AAV) capsid variants. The compositions and methods of the present disclosure are particularly useful for treating subjects suffering from, having been diagnosed with, or at risk of suffering from a GBA1-related disorder, such as Parkinson's disease (PD), Gaucher's disease (GD), Parkinson's disease dementia (PDD), dementia with Lewy bodies (DLB), or Lewy body dementia (LBD).

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202480029962.1 (22) Application Date 2024.05.01 (30) Priority Data 63 / 463,836 2023.05.03 US 63 / 593,792 2023.10.27 US 63 / 564,462 2024.03.12 US (85) PCT International Application Entering National Phase Date 2025.11.03 (86) PCT International Application Application Data PCT / US2024 / 027309 2024.05.01 (87) PCT International Application Publication Data WO2024 / 229161 EN 2024.11.07 (71) Applicant: Voyager Therapeutics, Inc. Address: Massachusetts, USA (72) Inventors: M.E. Nonnenmacher, T.C. Moyer, Li Jiangyu, D.R. Lax, Zhong Xiaoxin, T. Carter, E. Noel (74) Patent Agency: Longtian Intellectual Property Agency Co., Ltd. 72003 Patent Attorney: Fu Wenchuan (51) Int.Cl. C12N 15 / 86 (2006.01) (54) Invention Title: Compositions and methods for treating conditions associated with glucosylsphingosine monophosphate (GABA) β1 deficiency (57) Abstract: This disclosure relates to compositions and methods for delivering, particularly altering, for example enhancing, the content of GBA1 protein, via the use of an adeno-associated virus (AAV) capsid variant. The compositions and methods disclosed herein are particularly suitable for treating subjects who have, have been diagnosed with, or are at risk of developing GBA1-related conditions such as Parkinson's disease (PD), Gaucher disease (GD), Parkinson's dementia (PDD), dementia with Lewy bodies (DLB), or Lewy body dementia (LBD). Claims 21 pages Description 390 pages Sequence Listing (Electronic Publication) Drawings 30 pages CN 121127597 A 2025.12.12 CN 1 21 12 75 97 A 1. An adeno-associated virus (AAV) particle comprising: a) 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 an amino acid sequence of SPH; and (iii) [N3] comprises X4, X5, and X6, wherein at least one of X4, X5, or X6 is a basic amino acid; and b) a viral genome comprising a β-glucocerebrosidase 1 (GBA1) coding sequence.2. The AAV particle of claim 1, wherein the amino acid sequence [N1]-[N2]-[N3] is in the hypervariable ring IV of the AAV capsid variant. 3. The AAV particle of claim 1 or claim 2, wherein the AAV capsid variant is an AAV9 capsid variant. 4. The AAV particle of any one of claims 1 to 3, wherein [N1] comprises X1, X2, and X3, wherein at least one of X1, X2, or X3 is G. 5. The AAV particle of any one of claims 1 to 4, wherein [N2]-[N3] comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941). 6. An adeno-associated virus (AAV) particle comprising: a viral genome containing a β-glucocerebrosidase 1 (GBA1) coding sequence; and an AAV9 capsid variant containing the amino acid sequence of SPHSKA (SEQ ID NO: 941). 7. The AAV particle of claim 6, wherein the amino acid sequence of SPHSKA (SEQ ID NO: 941) is in the hypervariable ring IV of the AAV9 capsid variant. 8. The AAV particle of claim 6 or claim 7, wherein the amino acid sequence of SPHSKA (SEQ ID NO: 941) is immediately following the amino acid position corresponding to position 455 of SEQ ID NO: 4 or SEQ ID NO: 36. 9. The AAV particle of any one of claims 6 to 8, wherein the AAV9 capsid variant further comprises one, two, or all of the following: N at the amino acid position corresponding to position 452 of SEQ ID NO: 4, E at the amino acid position corresponding to position 451 of SEQ ID NO: 4, and / or V at the amino acid position corresponding to position 453 of SEQ ID NO: 4. 10. The AAV particle of any one of claims 6 to 9, wherein the AAV9 capsid variant comprises the amino acid sequence KTENVSGSPHSKAQNQQT (SEQ ID NO: 3272). 11. The AAV particle of any one of claims 6 to 10, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence having at least 90% identity with SEQ ID NO: 4; (ii) a VP2 protein comprising an amino acid sequence having at least 90% identity with positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein comprising an amino acid sequence having at least 90% identity with positions 203-742 of SEQ ID NO: 4.12. The AAV particle according to any one of claims 6 to 11, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence having at least 95% identity with SEQ ID NO: 4; (ii) a VP2 protein comprising an amino acid sequence having at least 95% identity with positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein comprising an amino acid sequence having at least 95% identity with positions 203-742 of SEQ ID NO: 4. 13. The AAV particle of any one of claims 6 to 12, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence having at least 99% identity with SEQ ID NO: 4; (ii) a VP2 protein comprising an amino acid sequence having at least 99% identity with positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein comprising an amino acid sequence having at least 99% identity with positions 203-742 of SEQ ID NO: 4. 14. The AAV particle of any one of claims 6 to 13, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 4; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 4. 15. The AAV particle of any one of claims 6 to 13, wherein the AAV9 capsid variant comprises: (i) the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately after the amino acid position corresponding to position 455 of SEQ ID NO: 4; (ii) E at the amino acid position corresponding to position 451 of SEQ ID NO: 4 and V at the amino acid position corresponding to position 453 of SEQ ID NO: 4; and (iii) no other modifications relative to wild-type AAV9. 16. The AAV particle of any one of claims 6 to 8, wherein the AAV9 capsid variant further comprises one, two, or all of the following: E ​​at the amino acid position corresponding to position 451 of SEQ ID NO: 36, R at the amino acid position corresponding to position 452 of SEQ ID NO: 36, and / or V at the amino acid position corresponding to position 453 of SEQ ID NO: 36.17. The AAV particle of any one of claims 6 to 8 and 16, wherein the AAV9 capsid variant comprises the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589). 18. The AAV particle of any one of claims 6 to 8, 16 and 17, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence having at least 90% identity with SEQ ID NO: 36; (ii) a VP2 protein comprising an amino acid sequence having at least 90% identity with positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein comprising an amino acid sequence having at least 90% identity with positions 203-742 of SEQ ID NO: 36. 19. The AAV particle according to any one of claims 6 to 8 and 16 to 18, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence having at least 95% identity with SEQ ID NO: 36; (ii) a VP2 protein comprising an amino acid sequence having at least 95% identity with positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein comprising an amino acid sequence having at least 95% identity with positions 203-742 of SEQ ID NO: 36. 20. The AAV particle according to any one of claims 6 to 8 and 16 to 19, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence having at least 99% identity with SEQ ID NO: 36; (ii) a VP2 protein comprising an amino acid sequence having at least 99% identity with positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein comprising an amino acid sequence having at least 99% identity with positions 203-742 of SEQ ID NO: 36. 21. The AAV particle of any one of claims 6 to 8 and 16 to 20, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 36; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 36. 22. The AAV particle of any one of claims 6 to 8 and 16 to 20, wherein the AAV9 capsid variant comprises: (i) an amino acid sequence SPHSKA (SEQ ID NO:941), wherein the amino acid sequence is present immediately after the amino acid position corresponding to position 455 of SEQ ID NO: 36; (ii) E at the amino acid position corresponding to position 451 of SEQ ID NO: 36, R at the amino acid position corresponding to position 452 of SEQ ID NO: 36, and V at the amino acid position corresponding to position 453 of SEQ ID NO: 36; and (iii) without any other modifications relative to wild-type AAV9. 23. The AAV particle of any one of claims 1 to 4, wherein [N1]-[N2]-[N3] are immediately following the position corresponding to amino acid position 452 of SEQ ID NO: 982; and wherein the AAV capsid variant comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 982, for example, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence at positions 203-742 of SEQ ID NO: 982. 24. The AAV particle of claim 23, wherein [N1] comprises GHD. 25. The AAV particle of claim 23 or claim 24, wherein [N1] comprises amino acid G at position 453 of SEQ ID NO: 138 or SEQ ID NO: 982, amino acid H at position 454 of SEQ ID NO: 138 or SEQ ID NO: 982, and amino acid D at position 455. 26. The AAV particle of any one of claims 23 to 25, wherein [N3] comprises KSG. 27. The AAV particle of any one of claims 23 to 26, wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity with SEQ ID NO: 982; (ii) a VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity with positions 138-742 of SEQ ID NO: 982; or (iii) a VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity with positions 203-742 of SEQ ID NO: 982. 28. The AAV particle of any one of claims 23 to 27, wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity with SEQ ID NO: 982.(ii) VP2 protein containing the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or having at least 95% identity with the amino acid sequence at positions 138-742 of SEQ ID NO: 982; or (iii) VP3 protein containing the amino acid sequence at positions 203-742 of SEQ ID NO: 982 or having at least 95% identity with the amino acid sequence at positions 203-742 of SEQ ID NO: 982. 29. The AAV particle according to any one of claims 23 to 28, wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity with SEQ ID NO: 982; (ii) a VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity with positions 138-742 of SEQ ID NO: 982; or (iii) a VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity with positions 203-742 of SEQ ID NO: 982. 30. The AAV particle of any one of claims 23 to 29, wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982; (ii) a VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 982; or (iii) a VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 982. 31. The AAV particle of any one of claims 1 to 30, wherein the viral genome encodes the wild-type GBA1 protein. 32. The AAV particle of any one of claims 1 to 31, wherein the viral genome does not encode a hemagglutinin (HA) tag. 33. The AAV particle of any one of claims 1 to 32, wherein the viral genome encodes a human GBA1 protein, a canine GBA1 protein, or a horse GBA1 protein. 34. The AAV particle of claim 33, wherein the viral genome encodes a human GBA1 protein, optionally wherein the human GBA1 protein comprises the amino acid sequence of SEQ ID NO: 1775. 35. The AAV particle of any one of claims 1 to 34, wherein the GBA1 coding sequence comprises the amino acid sequence of SEQ ID NO:36. The AAV particle of any one of claims 1 to 35, wherein the GBA1 coding sequence comprises a nucleotide sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 2002. 37. The AAV particle of any one of claims 1 to 36, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002. 38. The AAV particle of any one of claims 1 to 37, wherein the GBA1 coding sequence encodes a signal sequence comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 2005. 39. The AAV particle of claim 38, wherein the signal sequence comprises the amino acid sequence of SEQ ID NO: 2005. 40. The AAV particle of any one of claims 1 to 39, wherein the GBA1 coding sequence comprises a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 2001. 41. The AAV particle of any one of claims 40, wherein the GBA1 coding sequence comprises SEQ ID NO: 2001. 42. The AAV particle of any one of claims 1 to 41, wherein the viral genome further comprises a miRNA (miR) binding site that regulates the expression of the encoded GBA1 protein in cells or tissues of the DRG, liver, heart, hematopoietic lineage, or combinations thereof. 43. The AAV particle of any one of claims 1 to 42, wherein the viral genome further comprises a promoter effectively linked to the GBA1 coding sequence. 44. The AAV particle of claim 43, wherein the promoter is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to the nucleotide sequence of SEQ ID NO: 1834. 46. The AAV particle of any one of claims 1 to 45, wherein the viral genome further comprises an enhancer.47. The AAV particle of claim 46, wherein the enhancer comprises a CMV immediate early (CMVie) enhancer. 48. The AAV particle of claim 47, wherein the CMVie enhancer is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to the nucleotide sequence of SEQ ID NO: 1831. 49. The AAV particle of claim 47, wherein the CMVie enhancer comprises the nucleotide sequence of SEQ ID NO: 1831. 50. The AAV particle of any one of claims 1 to 49, wherein the viral genome further comprises introns. 51. The AAV particle of claim 50, wherein the intron is at least 90% identical to the nucleotide sequence of SEQ ID NO: 1842 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical). 52. The AAV particle of claim 50, wherein the intron comprises the nucleotide sequence of SEQ ID NO: 1842. 53. The AAV particle of any one of claims 1 to 52, wherein the viral genome further comprises a polyadenylated (polyadenylated) region. 54. The AAV particle of claim 53, wherein the polyadenylated region is at least 90% identical to the nucleotide sequence of SEQ ID NO: 1846 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical). 55. The AAV particle of claim 53, wherein the polyadenylated region comprises the nucleotide sequence of SEQ ID NO: 1846. 56. The AAV particle of any one of claims 1 to 55, wherein the viral genome further comprises an inverted terminal repeat (ITR). 57. The AAV particle of claim 56, wherein the ITR is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to the nucleotide sequence of SEQ ID NO: 1829 or SEQ ID NO: 1830. 58. The AAV particle of claim 56, wherein the ITR comprises the nucleotide sequence of SEQ ID NO: 1829 or SEQ ID NO: 1830. 59. The AAV particle of claim 56, wherein the viral genome comprises a 5' ITR and a 3' ITR.ITR, wherein the 5' ITR contains the nucleotide sequence of SEQ ID NO: 1829 and the 3' ITR contains the nucleotide sequence of SEQ ID NO: 1830. 60. The AAV particle of any one of claims 1 to 59, wherein the viral genome further contains one or more miR183 binding sites. 61. The AAV particle of claim 60, wherein the viral genome contains four miR183 binding sites. 62. The AAV particle of claim 61, wherein each of the four miR183 binding sites contains a nucleotide sequence that is at least 70% identical to the nucleotide sequence of SEQ ID NO: 1847, optionally, wherein each of the four miR183 binding sites contains the nucleotide sequence of SEQ ID NO: 1847. 63. The AAV particle of any one of claims 1 to 60, wherein the viral genome further comprises a series of miR183 binding sites comprising at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) of the nucleotide sequence of SEQ ID NO: 1849. 64. The AAV particle of claim 63, wherein the series of miR183 binding sites comprises the nucleotide sequence of SEQ ID NO: 1849. 65. The AAV particle of any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR); (ii) a promoter; (iii) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; and (iv) a 3' ITR. 66. The AAV particle of any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR); (ii) a promoter; (iii) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; and (iv) a 3' ITR. 67. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises: (i) a 5' inverted terminal repeat (ITR); (ii) a CMV immediate early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or the same as SEQ ID NO:(iii) A chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) a GBA1 coding sequence comprising the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; and (vi) 3' ITR. 68. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises: (i) a 5' inverted terminal repeat (ITR); (ii) a CMV immediate early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a chicken β-actin (CBA) promoter comprising the nucleotide sequence SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (iv) an intron comprising the nucleotide sequence SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; 1842 nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and (v) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; and (vi) a 3' ITR. 69. The AAV particle of any one of claims 1 to 34, wherein the viral genome comprises: (i) a 5' inverted terminal repeat (ITR); (ii) a CMV immediate early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO:(iii) A chicken β-actin (CBA) promoter comprising a nucleotide sequence of SEQ ID NO: 1831 or at least 95% identical to SEQ ID NO: 1831; (iv) an intron comprising a nucleotide sequence of SEQ ID NO: 1842 or at least 95% identical to SEQ ID NO: 1842 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%); (v) a GBA1 coding sequence comprising a nucleotide sequence of SEQ ID NO: 2002 or at least 90% identical to SEQ ID NO: 2002 (e.g., at least 93%); (vi) a polyadenylated (polyadenylated) region comprising SEQ ID NO: 1831. The nucleotide sequence of NO: 1846 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1846; and (vii) 3' ITR. 70. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR); (ii) a CMV immediate early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a chicken β-actin (CBA) promoter comprising the nucleotide sequence SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; and (iv) an intron comprising the nucleotide sequence SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834. (v) A nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 2001; (vi) A GBA1 coding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (vi) A polyadenylated (polyadenylated) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1846; and (vii) 3'ITR. 71. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) a CMV immediate early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (Claims 7 / 21, page 8, CN 121127597, A NO:) (iv) A nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 1834; (v) An intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 1842; (v) A GBA1 coding sequence comprising the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93%) to SEQ ID NO: 2002; (vi) A polyadenylated (polyadenylated) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 1846; and (vii) 3' ITR, comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1830. 72. The AAV particle of any one of claims 1 to 34, wherein the viral genome comprises: (i) a 5' inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1829 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (ii) a CMV immediate early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1829.(iii) A chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (iv) An intron comprising the nucleotide sequence of SEQ ID NO: 1842 or at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) A GBA1 coding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (vi) The AAV particle comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1846; and (vii) the 3' ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1830. 73. The AAV particle of any one of claims 1 to 34, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 97% identical) to SEQ ID NO: 2006. 74. The AAV particle of claim 73, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2006. 75. The AAV particle of claim 74, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2006. 76. The AAV particle of any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR); (ii) a promoter; (iii) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (iv)At least one miR183 binding site; and (v) a 3' ITR. 77. The AAV particle of any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR); (ii) a promoter; (iii) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (iv) at least one miR183 binding site; and (v) a 3' ITR. 78. The AAV particle of any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR); (ii) a promoter; (iii) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (iv) at least one miR183 binding site sequence comprising at least one miR183 binding site and at least one spacer sequence; and (v) a 3' ITR. 79. The AAV particle of any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR); (ii) a promoter; (iii) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (iv) at least one miR183 binding site sequence comprising at least one miR183 binding site and at least one spacer sequence; and (v) a 3' ITR. 80. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR); (ii) a CMV immediate early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a chicken β-actin (CBA) promoter comprising the nucleotide sequence SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; and (iv) an intron comprising the SEQ ID NO: 1834.(v) A nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (vi) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises a nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (vii) At least one miR183 binding site; and (vii) 3' ITR. 81. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR); (ii) a CMV immediate early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1842; (v) The GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (vi) at least one miR183 binding site; and (vii) a 3' ITR. 82. The AAV particle of any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR); (ii) a CMV immediate early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; and (iii) a chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831;(iv) A nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 1842; (v) An intron comprising a nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 1842; (vi) A GBA1 coding sequence comprising a nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93%) to SEQ ID NO: 2002; (vi) A miR183 binding site series comprising a nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical to SEQ ID NO: 1849; and (vii) A 3' ITR. 83. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR); (ii) a CMV immediate early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a chicken β-actin (CBA) promoter comprising the nucleotide sequence SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (iv) an intron comprising the nucleotide sequence SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (v) A nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 2001; (vi) A series of miR183 binding sites that contain the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 1849; and (vii)3' ITR. 84. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) a CMV immediate early (CMVie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: (iv) A nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (v) An intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (vi) A GBA1 coding sequence comprising the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (vi) At least one miR183 binding site comprising the nucleotide sequence of SEQ ID NO: 1847; (vii) A polyadenylated (polyadenylated) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834. 1846 nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and (viii) 3' ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1830. 85. The AAV particle of any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1830.(ii) A CMV immediate early (CMVie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence of SEQ ID NO: 1831 that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iii) A chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence of SEQ ID NO: 1834 that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) An intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence of SEQ ID NO: 1842 that is at least 95% identical. (See claims 11 / 21, page 12, CN) 121127597 A nucleotide sequence that is (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (v) a GBA1 coding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (vi) at least one miR183 binding site comprising the nucleotide sequence of SEQ ID NO: 1847; (vii) a polyadenylated (polyadenylated) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1846; and (viii) a 3' ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or at least 95% identical to SEQ ID NO: 1830. The same (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) nucleotide sequence. 86. The AAV particle of any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) a CMV immediate early (CMVie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO:(iii) A chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) a GBA1 coding sequence comprising the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (vi) The miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical to SEQ ID NO: 1849 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (vii) a polyadenylated (polyadenylated) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1846 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and (viii) a 3' ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1830 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical). 87. The AAV particle of any one of claims 1 to 34, wherein the viral genome comprises, in 5' to 3' order: (i) a 5' inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) a CMV immediate early (CMVie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii)Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) a GBA1 coding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (vi) a miR183 binding site series comprising SEQ ID NO: (i) a nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 1849; (vii) a polyadenylated (polyadenylated) region comprising a nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 1846; and (viii) a 3' ITR comprising a nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 1830. 88. The AAV particle of claim 87, wherein: (i) the 5' ITR comprises the nucleotide sequence of SEQ ID NO: 1829; (ii) the CMVie enhancer comprises the nucleotide sequence of SEQ ID NO: 1831; (iii) the CBA promoter comprises the nucleotide sequence of SEQ ID NO: 1834; (iv) the intron comprises the nucleotide sequence of SEQ ID NO: 1842; (v) the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001; (vi) the miR183 binding site sequence comprises the nucleotide sequence of SEQ ID NO: 1849; (vii) the polyadenylated region comprises the nucleotide sequence of SEQ ID NO: 1846; and (viii) the 3'The ITR contains the nucleotide sequence of SEQ ID NO: 1830. 89. The AAV particle of any one of claims 1 to 34, wherein the viral genome contains the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 97% identical) to SEQ ID NO: 2007. 90. The AAV particle of claim 89, wherein the viral genome contains the nucleotide sequence of SEQ ID NO: 2007. 91. The AAV particle of claim 90, wherein the viral genome consists of the nucleotide sequence of SEQ ID NO: 2007. 92. An adeno-associated virus (AAV) particle comprising a viral genome containing the nucleotide sequence of SEQ ID NO: 2001 and an AAV capsid variant comprising: (i) a VP1 protein containing the amino acid sequence of SEQ ID NO: 4; (ii) a VP2 protein containing the amino acid sequence of positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein containing the amino acid sequence of positions 203-742 of SEQ ID NO: 4. 93. An adeno-associated virus (AAV) particle comprising a viral genome containing the nucleotide sequence of SEQ ID NO: 2001 and an AAV capsid variant comprising: (i) a VP1 protein containing the amino acid sequence of SEQ ID NO: 36; (ii) a VP2 protein containing the amino acid sequence of positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein containing the amino acid sequence of positions 203-742 of SEQ ID NO: 36. Claims 13 / 21, page 14, CN 121127597 A 94. An adeno-associated virus (AAV) particle comprising a viral genome containing the nucleotide sequence of SEQ ID NO: 2002 and an AAV capsid variant comprising: (i) a VP1 protein containing the amino acid sequence of SEQ ID NO: 4; (ii) a VP2 protein containing the amino acid sequence at positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein containing the amino acid sequence at positions 203-742 of SEQ ID NO: 4. 95. An adeno-associated virus (AAV) particle comprising a viral genome containing the nucleotide sequence of SEQ ID NO: 2002 and an AAV capsid variant comprising: (i) a VP1 protein containing the amino acid sequence of SEQ ID NO: 36; (ii) a VP2 protein containing the amino acid sequence at positions 138-742 of SEQ ID NO: 36; and / or(iii) VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 36. 96. A cell comprising AAV particles as described in any one of claims 1 to 95, optionally wherein the cell is a mammalian cell (e.g., HEK293 cell), an insect cell (e.g., Sf9 cell), or a bacterial cell. 97. A method for preparing AAV particles as described in any one of claims 1 to 95, the method comprising: (i) providing a cell comprising a viral genome containing a GBA1 coding sequence and nucleic acid encoding the AAV capsid variant; and (ii) culturing the cell under conditions suitable for encapsulating the viral genome within the AAV capsid variant; thereby preparing the AAV particles. 98. The method of claim 97, wherein the viral genome comprises (a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); or (b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 4 or an amino acid sequence identical to SEQ ID NO: 2006. 4. An amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity); (ii) a VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 4 or having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) with the amino acid sequence at positions 138-742 of SEQ ID NO: 4; or (iii) a VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 4 or having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) with the amino acid sequence at positions 203-742 of SEQ ID NO: 4. Positions 203-742 have an amino acid sequence with at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity).99. The method of claim 97, wherein the viral genome comprises (a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); or (b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 4, the amino acid sequence at positions 138-742 of SEQ ID NO: 4, and / or the amino acid sequence at positions 203-742 of SEQ ID NO: 4. 100. The method of claim 97, wherein the viral genome comprises: (a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical thereto (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical thereto); or (b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical thereto (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical thereto); and wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence that is identical to or identical to that of SEQ ID NO: 2007. 36 has an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity); (ii) VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 36 or having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity); and / or (iii) VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 36 or having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity); and / or VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 36 or having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity).Position 36, 203-742, has an amino acid sequence with at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity). 101. The method of claim 97, wherein the viral genome comprises: (a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical thereto (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical thereto); or (b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical thereto (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical thereto); and wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 36, the amino acid sequence at positions 138-742 of SEQ ID NO: 36, and / or the amino acid sequence at positions 203-742 of SEQ ID NO: 36. 102. The method of claim 97, wherein the viral genome comprises: (a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical thereto (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical thereto); or (b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical thereto (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical thereto); and wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or having at least 90% identical to SEQ ID NO: 982. (Claims 15 / 21, Page 16, CN 121127597 A) (i) an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity; (ii) a VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) with respect to positions 138-742 of SEQ ID NO: 982.(iii) an amino acid sequence having at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity; and / or (iii) a VP3 protein comprising the amino acid sequence at position 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) with position 203-742 of SEQ ID NO: 982. 103. The method of claim 97, wherein the viral genome comprises: (a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical thereto (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical thereto); or (b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical thereto (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical thereto); and wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 982, the amino acid sequence at positions 138-742 of SEQ ID NO: 982, and / or the amino acid sequence at positions 203-742 of SEQ ID NO: 982. 104. The method of any one of claims 97 to 103, further comprising introducing a first nucleic acid molecule comprising the viral genome into the cell prior to step (i). 105. The method of any one of claims 97 to 104, wherein the cell comprises a second nucleic acid molecule encoding the AAV capsid variant. 106. The method of claim 105, further comprising introducing the second nucleic acid into the cell prior to step (i). 107. The method of any one of claims 97 to 106, wherein the cell comprises mammalian cells (e.g., HEK293 cells), insect cells (e.g., Sf9 cells), or bacterial cells. 108. A pharmaceutical composition comprising AAV particles as described in any one of claims 1 to 95 and a pharmaceutically acceptable excipient. 109. A pharmaceutical composition comprising AAV particles as described in any one of claims 5 to 22 and a pharmaceutically acceptable excipient. 110. A pharmaceutical composition comprising AAV particles as described in any one of claims 9 to 15, 92, and 94 and a pharmaceutically acceptable excipient. 111. A pharmaceutical composition comprising AAV particles and a drug substance as described in any one of claims 16 to 22, 93 and 95.Physically acceptable excipients. 112. A method of delivering AAV particles encoding GBA1 protein to a subject, comprising administering to the subject an effective amount of the pharmaceutical composition as described in any one of claims 108 to 111 or the AAV particles as described in any one of claims 1 to 95. 113. The method of claim 112, wherein the subject has, has been diagnosed with, or is at risk of developing a GBA1-related condition, optionally wherein the GBA1-related condition is a GBA1-related neurodegenerative or neuromuscular disease, further optionally wherein the GBA1-related condition is Parkinson's disease (PD), Parkinson's dementia (PDD), Gaucher disease (GD) (e.g., GD type 1, 2, or 3), dementia with Lewy bodies (DLB), Lewy body dementia (LBD), multiple system atrophy (MSA), Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), isolated autonomic failure, brain iron deposition neurodegeneration type 1 (NBIA 1), or Hallewarden-Schpattz syndrome. 114. The method of claim 112 or claim 113, wherein the subject has, has been diagnosed with, or is at risk of developing Parkinson's disease (PD). 115. A method of treating a subject who has or has been diagnosed with GBA1-related conditions, comprising administering to the subject an effective amount of the pharmaceutical composition of any one of claims 108 to 111 or AAV particles of any one of claims 1 to 95. 116. A method of treating a subject who has or has been diagnosed with GBA1-related conditions, comprising administering to the subject an effective amount of the pharmaceutical composition of any one of claims 109 to 111 or AAV particles of any one of claims 5 to 22. 117. A method of treating a subject who has or has been diagnosed with GBA1-related conditions, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 110 or AAV particles of any one of claims 9 to 15, 92, and 94. 118. A method of treating a subject suffering from or diagnosed with a GBA1-related condition, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 111 or AAV particles as described in any one of claims 16 to 22, 93, and 95. 119. The method of any one of claims 115 to 118, wherein the GBA1-related condition is a GBA1-related neurodegenerative or neuromuscular disease, optionally wherein the GBA1-related condition is Parkinson's disease (PD), Parkinson's dementia (PDD), or Gaucher disease (GD) (e.g., GD...).120. The method of claim 119, wherein the GBA1-related neurodegenerative disease or neuromuscular disorder is Parkinson's disease (PD). 121. The method of claim 119, wherein the GBA1-related neurodegenerative disease or neuromuscular disorder is Gaucher disease (GD) (e.g., GD type 1, 2, or 3). 122. The method of claim 121, wherein the GBA1-related neurodegenerative disease or neuromuscular disorder is GD type 1. 123. The method of claim 121, wherein the GBA1-related neurodegenerative disease or neuromuscular disorder is GD type 2. 124. The method of claim 121, wherein the GBA1-related neurodegeneration or neuromuscular disease is GD type 3. 125. The method of claim 119, wherein the GBA1-related neurodegeneration or neuromuscular disease is Lewy body dementia (DLB). 126. The method of claim 119, wherein the GBA1-related neurodegeneration or neuromuscular disease is Lewy body dementia (LBD). 127. A method of treating a subject suffering from or diagnosed with Parkinson's disease (PD), comprising administering to the subject an effective amount of a pharmaceutical composition as described in any one of claims 108 to 111 or AAV particles as described in any one of claims 1 to 95. Claims 17 / 21 pages 18 CN 121127597 A 128. A method of treating a subject suffering from or diagnosed with Parkinson's disease (PD), comprising administering to the subject an effective amount of a pharmaceutical composition as described in any one of claims 109 to 111 or AAV particles as described in any one of claims 5 to 22. 129. A method of treating a subject with Parkinson's disease (PD) or diagnosed with PD, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 110 or AAV particles as described in any one of claims 9 to 15, 92, and 94. 130. A method of treating a subject with Parkinson's disease (PD) or diagnosed with PD, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 111 or AAV particles as described in any one of claims 16 to 22, 93, and 95. 131. A method of treating a subject with Gaucher disease (GD) (e.g., GD type 1, 2, or 3) or diagnosed with Gaucher disease (GD) (e.g., GD...132. A method for treating a subject with Gaucher disease (GD) (e.g., GD type 1, 2, or 3) or diagnosed with Gaucher disease (GD) (e.g., GD type 1, 2, or 3), comprising administering to the subject an effective amount of the pharmaceutical composition as described in any one of claims 108 to 111 or AAV particles as described in any one of claims 1 to 22. 133. A method for treating a subject with Gaucher disease (GD) (e.g., GD type 1, 2, or 3) or diagnosed with Gaucher disease (GD) (e.g., GD type 1, 2, or 3), comprising administering to the subject an effective amount of the pharmaceutical composition as described in claims 110 or AAV particles as described in any one of claims 9 to 15, 92, and 94. 134. A method of treating a subject with Gaucher disease (GD) (e.g., GD type 1, 2, or 3) or diagnosed with Gaucher disease (GD) (e.g., GD type 1, 2, or 3), comprising administering to the subject an effective amount of the pharmaceutical composition of claim 111 or AAV particles as described in any one of claims 16 to 22, 93, and 95. 135. The method of any one of claims 131 to 134, wherein the GD is GD type 1. 136. The method of any one of claims 131 to 134, wherein the GD is GD type 2. 137. The method of any one of claims 131 to 134, wherein the GD is GD type 3. 138. A method of treating a subject with Lewy body dementia (LBD), comprising administering to the subject an effective amount of the pharmaceutical composition of any one of claims 108 to 111 or AAV particles as described in any one of claims 1 to 95. 139. A method of treating a subject with Lewy body dementia (LBD), comprising administering to the subject an effective amount of the pharmaceutical composition as described in claims 109 to 111 or AAV particles as described in any one of claims 5 to 22. 140. A method of treating a subject with Lewy body dementia (LBD), comprising administering to the subject an effective amount of the pharmaceutical composition as described in claim 111 or AAV particles as described in any one of claims 9 to 15, 92, and 94. 141. A method of treating a subject with Lewy body dementia (LBD), comprising administering to the subject an effective amount of the pharmaceutical composition as described in claim 111 or AAV particles as described in any one of claims 16 to 22, 93, and 95. 142. A method of treating a subject with dementia having a Lewy body (DLB), comprising administering to the subject an effective amount of the pharmaceutical composition as described in claim 111 or AAV particles as described in any one of claims 16 to 22, 93, and 95.An effective amount of the pharmaceutical composition as described in any one of claims 108 to 111 or AAV particles as described in any one of claims 1 to 95. 143. A method of treating a subject with Lewy body dementia (DLB), comprising administering to the subject an effective amount of the pharmaceutical composition as described in claims 109 to 111 or AAV particles as described in any one of claims 5 to 22. 144. A method of treating a subject with Lewy body dementia (DLB), comprising administering to the subject an effective amount of the pharmaceutical composition as described in claims 111 or AAV particles as described in any one of claims 9 to 15, 92, and 94. 145. A method of treating a subject with Lewy body dementia (DLB), comprising administering to the subject an effective amount of the pharmaceutical composition as described in claims 111 or AAV particles as described in any one of claims 16 to 22, 93, and 95. 146. The method of any one of claims 112 to 145, wherein the subject has one or more mutations in the GBA1 gene. 147. The method of any one of claims 112 to 146, wherein the subject has lower GC enzyme activity compared to the subject without GBA1-related disease, optionally wherein the level of GC enzyme activity is measured by a 4-MUG assay or a SensoLyte blue glucocerebrosidase assay. 148. The method of any one of claims 115 to 147, wherein the treatment halts the progression of the disease in the subject. 149. The method of any one of claims 115 to 148, wherein the treatment improves at least one symptom of the disease and / or alters one or more biomarkers of the disease. 150. The method of claim 149, wherein the one or more biomarkers comprise GC enzyme activity, levels of glucocerebrosidase and other glycolipids (e.g., in immune cells, such as macrophages), levels of synuclein aggregates (e.g., Lewy bodies), levels of neurofilament light chains, or combinations thereof. 151. The method of claim 149, wherein the at least one symptom comprises developmental delay, progressive encephalopathy, progressive dementia, ataxia, myoclonus, oculomotor dysfunction, bulbar palsy, general weakness, limb tremor, depression, visual hallucinations, cognitive decline, or a combination thereof. 152. The method of any one of claims 112 to 151, wherein the subject is a human. 153. The method of any one of claims 112 to 152, wherein the AAV particles are delivered to cells, tissues, or regions of the CNS, such as regions of the brain or spinal cord, such as the parenchyma, cortex, substantia nigra, caudate nucleus, cerebellum, striatum, corpus callosum, cerebellum, etc.154. The method of any one of claims 112 to 153, further comprising assessing, for example, measuring, in the subject, such as in the subject's cells, tissues, or body fluids, the level of GBA1 expression, such as the level of GBA1 gene expression, the level of GBA1 mRNA expression, and / or the level of GBA1 protein expression. 155. The method of any one of claims 112 to 154, wherein the level of GBA1 protein expression is measured by ELISA, Western blotting, or immunohistochemistry. 156. The method of claim 154 or claim 155, wherein the assessment of the level of GBA1 expression is performed before and after administration of the AAV, optionally, wherein the level of GBA1 expression before administration is compared with the level of GBA1 expression after administration. 157. The method of any one of claims 154 to 156, comprising assessing the level of GBA1 expression in the cells or tissues of the central nervous system (e.g., parenchyma). 158. The method of claim 156 or claim 157, wherein the subject's GBA1 protein expression level increases after administration relative to the subject's GBA1 protein expression level prior to administration. Claims 19 / 21 pages 20 CN 121127597 A 159. The method of any one of claims 112 to 158, further comprising assessment, for example, measuring the GC enzyme activity level in the subject. 160. The method of any one of claims 112 to 159, wherein the administration results in an increase of: (i) GC enzyme activity in the subject's cells, tissues (e.g., cells or tissues of the CNS, such as the cortex, striatum, thalamus, cerebellum, and / or brainstem) and / or body fluids (e.g., CSF and / or serum), optionally wherein the GC enzyme activity in the subject increases by at least 2-fold, at least 3-fold, at least 4-fold, or at least 5-fold after the administration relative to the GC enzyme activity in the subject before the administration; (ii) the level of viral genome (VG) in each cell of the subject's CNS tissues (e.g., the cortex, striatum, thalamus, cerebellum, brainstem, and / or spinal cord), optionally wherein the level of VG in each cell of the subject's CNS tissue increases by more than 50 VG per cell relative to the level of VG in each cell of the subject's peripheral tissues; and / or (iii) GBA1 mRNA expression in the subject's cells or tissues (e.g., cells or tissues of the CNS, such as the cortex, thalamus, and / or brainstem), optionally, wherein GBA1 mRNA expression in the subject after administration is higher than that before administration.mRNA expression increased by at least 100-1300-fold. 161. The method of any one of claims 115 to 160, further comprising administering to the subject an additional agent suitable for treating or preventing the GBA1-related condition; optionally, wherein the additional agent comprises enzyme replacement therapy (ERT) (e.g., imiglucerase, veralucerase α, or taliglucerase α); substrate reduction therapy (SRT) (e.g., elixstat or miglustat), levodopa, carbidopa, safenamide, dopamine agonists (e.g., quetiapine, clozapine, pramipexole, rotigotine, or ropinirole), anticholinergics (e.g., benzalkonium chloride or trihexyphenidyl), cholinesterase inhibitors (e.g., rivastigmine, donepezil, or galantamine), N-methyl-d-aspartate (NMDA) receptor antagonists (e.g., memantine), or combinations thereof. 162. The method of any one of claims 112 to 161, further comprising administering a blood transfusion to the subject. 163. The method of any one of claims 112 to 161, further comprising administering an immunosuppressant to the subject. 164. The method of claim 162, wherein the immunosuppressant comprises a corticosteroid (e.g., prednisone, prednisolone, methylprednisolone, and / or dexamethasone), rapamycin, mycophenolate mofetil, tacrolimus, rituximab, and / or ikulizumab hydroxychloroquine. 165. The pharmaceutical composition of any one of claims 108 to 111 or the AAV particles of any one of claims 1 to 95, for the treatment of GBA1-related conditions; optionally, wherein the GBA1-related condition is Parkinson's disease (PD), Parkinson's dementia (PDD), Gaucher disease (GD) (e.g., GD type 1, 2, or 3), Lewy body dementia (LBD), Lewy body dementia (LBD), multiple system atrophy (MSA), Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), isolated autonomic failure, brain iron deposition neurodegeneration type 1 (NBIA 1), or Hallewarden-Schpatrick syndrome. 166. The pharmaceutical composition or AAV particles of claim 165, wherein the GBA1-related condition is Gaucher disease (GD) (e.g., GD type 1, 2, or 3). 167. The pharmaceutical composition or AAV particles of claim 166, wherein the GD is GD type 1. 168. The pharmaceutical composition or AAV particles of claim 166, wherein the GD is GD type 2. 169. The pharmaceutical composition or AAV particles of claim 166, wherein the GD is GD type 3. 170. The pharmaceutical composition or AAV particles of claim 165, wherein the GBA1-related condition is Parkinson's disease (PD). Claims 20 / 21 pages 21 CN 121127597 A171. The pharmaceutical composition or AAV particles of claim 165, wherein the GBA1-related condition is LBD. 172. The pharmaceutical composition or AAV particles of claim 165, wherein the GBA1-related condition is DLB. 173. Use of a pharmaceutical composition of any one of claims 108 to 111 or an AAV particle of any one of claims 1 to 95 in the preparation of a medicament for treating GBA1-related conditions; optionally, wherein the GBA1-related condition is Parkinson's disease (PD), Parkinson's dementia (PDD), Gaucher disease (GD) (e.g., GD type 1, 2, or 3), dementia with Lewy bodies (DLB), Lewy body dementia (LBD), multiple system atrophy (MSA), Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), isolated autonomic failure, brain iron deposition neurodegeneration type 1 (NBIA 1), or Hallewarden-Schpattz syndrome. 174. The use of claim 173, wherein the GBA1-related condition is PD. 175. The use of claim 173, wherein the GBA1-related condition is Gaucher disease (GD) (e.g., GD type 1, 2, or 3). 176. The use of claim 175, wherein the GD is GD type 1. 177. The use of claim 175, wherein the GD is GD type 2. 178. The use of claim 175, wherein the GD is GD type 3. 179. The use of claim 173, wherein the GBA1-related condition is LBD. 180. The use of claim 173, wherein the GBA1-related condition is DLB. Claims 21 / 21 Page 22 CN 121127597 A Composition and method for treating conditions related to glucosylsphingosine monophosphate (GMP) deficiency

[0001] Related Applications

[0002] This application claims the benefit and priority of U.S. Provisional Application Serial No. 63 / 463,836, filed May 3, 2023; U.S. Provisional Application Serial No. 63 / 593,792, filed October 27, 2023; and U.S. Provisional Application Serial No. 63 / 564,462, filed March 12, 2024, the contents of each of which are incorporated herein by reference in their entirety.

[0003] Serial List

[0004] This application is filed with an electronic serial list. The serial list file, named 14640_0081-00304_SL.xml, was created on April 10, 2024, and is 5,682,716 bytes in size. Information from the electronic sequence listing is incorporated herein by reference in its entirety. Technical Field

[0005] This document describes compositions and methods relating to adeno-associated virus (AAV) viral particles used for…The composition delivers a polynucleotide, such as a polynucleotide encoding glucosylsphingosine monophosphate β1 (GBA1) protein (“GBA1 protein”) and a peptide (“GBA1 peptide”), for the treatment of GBA1-related conditions, including Parkinson's disease (PD) and other GBA1-related conditions, including Gaucher disease, Parkinson's dementia (PDD), dementia with Lewy bodies (DLB), and Lewy body dementia (LBD). In some embodiments, the compositions described herein may be used to treat subjects in need, such as human subjects diagnosed with a GBA1-related condition or other conditions resulting from a deficiency in the quantity and / or function of the GBA1 protein.

[0006] Prior Art

[0007] Lysosomal acid glucosylceramidase, commonly known as glucosylcerebrosidase or GC enzyme, is a D-glucosyl-N-acylsphingosine glucohydrolase, a lysosomal membrane protein that plays an important role in glucose and lipid metabolism. This enzyme is encoded by the glucosylceramidase β (GBA1) gene (Ensembl gene ID ENSG00000177628). This enzyme, along with Saposin A and Saposin C, catalyzes the hydrolysis of glucose-ceramide into ceramide and glucose. See Vaccaro, Anna Maria et al., Journal of Biological Chemistry 272.27 (1997): 16862-16867.

[0008] Mutations in GBA1 are known to cause disease in human subjects. Homozygous or compound heterozygous GBA1 mutations result in Gaucher disease (“GD”). See Sardi, S. Pablo, Jesse M. Cedarbaum, and Patrik Brundin. Movement Disorders 33.5 (2018): 684-696. Gaucher disease is one of the most common lysosomal storage diseases, with a standardized birth incidence estimated at 0.4 to 5.8 per 100,000 subjects in the general population. Heterozygous GBA1 mutations may lead to PD. In fact, the mutation rate of GBA1 in all PD patients is 7%-10%, making GBA1 mutations a significant risk factor.The most important genetic risk factor for PD. PD-GBA patients have reduced levels of the lysosomal enzyme β-glucocerebrosidase (GC enzyme), leading to increased accumulation of glycosphingolipid glucosylceramide (GluCer), which in turn is associated with increased α-synuclein aggregation and accompanying neurological symptoms. GD and PD are associated with other lysosomal storage diseases or Lewy body diseases, such as Lewy body dementia (LBD). See Sidransky, E. and Lopez, G. Lancet Neurol. Product Manual 1 / 390 pages 23 CN 121127597 A Nov 2012; 11(11): 986-998.

[0009] To date, treatments available for GBA1-related conditions such as PD are limited, and delivery to the adult central nervous system (CNS) remains a major challenge in the development of new and effective therapies. Therefore, there remains a long-standing need to develop pharmaceutical compositions and methods that can be delivered to the CNS for the treatment of PD and other GBA1-related conditions and to improve the deficiency of GBA1 protein in subjects (e.g., human subjects) suffering from GBA1-related conditions.

[0010] Adeno-associated virus (AAV) has been presented as a widely studied and utilized viral particle for the delivery of therapeutically effective peptides to mammalian cells. See, for example, Tratschin et al., Mol. Cell Biol., 5(11):3251-3260 (1985) and Grimm et al., Hum. Gene Ther., 10(15):2445-2450 (1999). This disclosure provides improved pharmaceutical compositions and methods. In some embodiments, this disclosure provides treatment methods using AAV capsid variants capable of delivering GBA1 to target cells or tissues (e.g., CNS cells or tissues). Summary of the Invention

[0011] This disclosure addresses these challenges by providing AAV-based compositions and methods containing AAV capsid variants for the treatment of GC enzyme deficiency in subjects. This document discloses compositions and methods relating to AAV-based gene delivery of GC enzymes to improve loss of function and improve intracellular lipid migration. These compositions and methods are applicable to improving lysosomal glycolipid metabolism in subjects (e.g., subjects with at least one mutation in the GBA1 gene), and to slowing, halting, or reversing neurodegenerative and other symptoms of PD and other GBA1-related conditions (e.g., Lewy body dementia (DLB), Gaucher disease (GD)). Unless otherwise stated, GBA protein, GBA1 protein, and GC enzyme protein are synonymous and used interchangeably to refer to the protein encoded by the GBA1 gene. Unless otherwise stated, the nucleotide sequence encoding the GBA1 protein (i.e., the GBA1 coding sequence) refers to...The nucleotide sequence encoding the amino acids of the GBA1 protein, and may also be referred to as the GBA1 protein coding sequence.

[0012] In some embodiments, this disclosure provides an AAV particle comprising a nucleotide sequence encoding a GBA1 protein (e.g., wild-type GBA1 protein) and an AAV capsid (e.g., an AAV capsid variant). In some embodiments, this disclosure provides an AAV particle comprising a nucleotide sequence encoding a wild-type GBA1 protein and an AAV capsid variant. In some embodiments, compared to the wild-type GBA1 coding sequence (e.g., a nucleotide sequence comprising SEQ ID NO: 1776 or 1777), the GBA1 coding nucleotide sequence comprises altered GC content and / or a reduced number of CpG motifs (e.g., lack of all CpG motifs), and wherein the AAV capsid variant is an AAV9 capsid variant.

[0013] In some embodiments, the AAV capsid variant is an AAV9 capsid variant that contains a peptide insert in the ring IV region. In some embodiments, the AAV capsid variant contains the amino acid sequence of SPH in the ring IV region. In some embodiments, the AAV capsid variant contains an amino acid sequence of SPH in ring IV, wherein the amino acid sequence (SPH) is present immediately after position 455 according to SEQ ID NO: 138.

[0014] In some embodiments, this disclosure provides an adeno-associated virus (AAV) particle containing an AAV capsid variant having an amino acid sequence of the formula [N1]-[N2]-[N3], wherein: optionally, [N1] contains X1, X2, and X3, wherein at least one of X1, X2, or X3 is G; wherein [N2] contains the amino acid sequence of SPH; and wherein [N3] contains X4, X5, and X6, wherein at least one of X4, X5, or X6 is a basic amino acid; wherein the AAV particle further contains a viral genome containing a β-glucocerebrosidase 1 (GBA1) coding sequence. In some embodiments, the amino acid sequence [N1]-[N2]-[N3] is located in the hypervariable loop IV of the AAV capsid variant. In some embodiments, the AAV capsid variant is the AAV9 capsid variant. In some embodiments, [N1] comprises X1, X2, and X3, wherein at least one of X1, X2, or X3 is G. In some embodiments, [N2]-[N3] comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941).

[0015] In some embodiments, this disclosure provides an AAV particle comprising: a genome containing the GBA1 coding sequence (see Virus Specification 2 / 390 pages 24 CN 121127597 A) and an AAV9 capsid variant containing the amino acid sequence of SPHSKA (SEQ ID NO: 941). In some embodimentsIn this embodiment, the amino acid sequence of SPHSKA (SEQ ID NO: 941) is located in the hypervariable ring IV of the AAV9 capsid variant. In some embodiments, the amino acid sequence of SPHSKA (SEQ ID NO: 941) is present immediately after the amino acid position corresponding to position 455 of SEQ ID NO: 4 or SEQ ID NO: 36.

[0016] In some embodiments, the AAV9 capsid variant comprises one, two, or all of the following: N at the amino acid position corresponding to position 452 of SEQ ID NO: 4, E at the amino acid position corresponding to position 451 of SEQ ID NO: 4, and / or V at the amino acid position corresponding to position 453 of SEQ ID NO: 4. In some embodiments, the AAV9 capsid variant comprises the amino acid sequence KTENVSGSPHSKAQNQQT (SEQ ID NO: 3272).

[0017] In some embodiments, the AAV9 capsid variant comprises a VP1 protein containing an amino acid sequence having at least 90% identity with SEQ ID NO: 4; a VP2 protein containing an amino acid sequence having at least 90% identity with positions 138-742 of SEQ ID NO: 4; and / or a VP3 protein containing an amino acid sequence having at least 90% identity with positions 203-742 of SEQ ID NO: 4. In some embodiments, the AAV9 capsid variant comprises a VP1 protein containing an amino acid sequence having at least 95% identity with SEQ ID NO: 4; a VP2 protein containing an amino acid sequence having at least 95% identity with positions 138-742 of SEQ ID NO: 4; and / or a VP3 protein containing an amino acid sequence having at least 95% identity with positions 203-742 of SEQ ID NO: 4. In some embodiments, the AAV9 capsid variant comprises a VP1 protein containing an amino acid sequence having at least 99% identity with SEQ ID NO: 4; a VP2 protein containing an amino acid sequence having at least 99% identity with positions 138-742 of SEQ ID NO: 4; and / or a VP3 protein containing an amino acid sequence having at least 99% identity with positions 203-742 of SEQ ID NO: 4. In some embodiments, the AAV9 capsid variant comprises a VP1 protein containing the amino acid sequence of SEQ ID NO: 4; a VP2 protein containing the amino acid sequence at positions 138-742 of SEQ ID NO: 4; and / or a VP3 protein containing the amino acid sequence at positions 203-742 of SEQ ID NO: 4.

[0018] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately following the amino acid position corresponding to position 455 of SEQ ID NO: 4; an E at the amino acid position corresponding to position 451 of SEQ ID NO: 4 and a V at the amino acid position corresponding to position 453 of SEQ ID NO: 4; and no other modifications relative to wild-type AAV9.

[0019] In some embodiments, the AAV9 capsid variant comprises one, two, or all of the following: an E at the amino acid position corresponding to position 451 of SEQ ID NO: 36, an R at the amino acid position corresponding to position 452 of SEQ ID NO: 36, and / or a V at the amino acid position corresponding to position 453 of SEQ ID NO: 36. In some embodiments, the AAV9 capsid variant comprises the amino acid sequence KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589).

[0020] In some embodiments, the AAV9 capsid variant comprises a VP1 protein containing an amino acid sequence having at least 90% identity with SEQ ID NO: 36; a VP2 protein containing an amino acid sequence having at least 90% identity with positions 138-742 of SEQ ID NO: 36; and / or a VP3 protein containing an amino acid sequence having at least 90% identity with positions 203-742 of SEQ ID NO: 36. In some embodiments, the AAV9 capsid variant comprises a VP1 protein containing an amino acid sequence having at least 95% identity with SEQ ID NO: 36; a VP2 protein containing an amino acid sequence having at least 95% identity with positions 138-742 of SEQ ID NO: 36; and / or a VP3 protein containing an amino acid sequence having at least 95% identity with positions 203-742 of SEQ ID NO: 36. In some embodiments, the AAV9 capsid variant comprises a VP1 protein containing an amino acid sequence having at least 99% identity with SEQ ID NO: 36; a VP2 protein containing an amino acid sequence having at least 99% identity with positions 138-742 of SEQ ID NO: 36; and / or a VP3 protein containing an amino acid sequence having at least 99% identity with positions 203-742 of SEQ ID NO: 36. In some embodiments, the AAV9 capsid variant comprises a VP1 protein containing the amino acid sequence of SEQ ID NO: 36; a VP2 protein containing the amino acid sequence of SEQ ID NO: 36; and / or a VP3 protein containing the amino acid sequence of SEQ ID NO: 36. (See specification 3 / 390 pages 25 CN 121127597 A)The amino acid sequence at positions 138-742; and / or the VP3 protein, which comprises the amino acid sequence at positions 203-742 of SEQ ID NO: 36.

[0021] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately after the amino acid position corresponding to position 455 of SEQ ID NO: 36; E at the amino acid position corresponding to position 451 of SEQ ID NO: 36, R at the amino acid position corresponding to position 452 of SEQ ID NO: 36, and V at the amino acid position corresponding to position 453 of SEQ ID NO: 36; and no other modifications relative to wild-type AAV9.

[0022] In some embodiments, the AAV capsid variant (e.g., the AAV9 capsid variant) comprises [N1]-[N2]-[N3] immediately following the position corresponding to amino acid position 452 of SEQ ID NO: 982, wherein the AAV capsid variant comprises an amino acid sequence that is at least 90% identical to, for example, positions 203-742 of SEQ ID NO: 982, such as at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical. In some embodiments, [N1] comprises GHD. In some embodiments, [N1] comprises amino acid G at position 453 of SEQ ID NO: 138 or SEQ ID NO: 982, amino acid H at position 454 of SEQ ID NO: 138 or SEQ ID NO: 982, and amino acid D at position 455 of SEQ ID NO: 138 or SEQ ID NO: 982. In some embodiments, [N3] comprises KSG.

[0023] In some embodiments, the AAV capsid variant comprises a VP1 protein containing an amino acid sequence having at least 90% identity with SEQ ID NO: 982; a VP2 protein containing an amino acid sequence having at least 90% identity with positions 138-742 of SEQ ID NO: 982; and / or a VP3 protein containing an amino acid sequence having at least 90% identity with positions 203-742 of SEQ ID NO: 982. In some embodiments, the AAV capsid variant comprises a VP1 protein having an amino acid sequence having at least 95% identity with SEQ ID NO: 982; and a VP2 protein having the same position as SEQ ID NO: 982.The AAV capsid variant comprises an amino acid sequence having at least 95% identity with SEQ ID NO: 982, specifically positions 138-742; and / or a VP3 protein containing an amino acid sequence having at least 95% identity with positions 203-742 of SEQ ID NO: 982. In some embodiments, the AAV capsid variant comprises a VP1 protein containing an amino acid sequence having at least 99% identity with SEQ ID NO: 982; a VP2 protein containing an amino acid sequence having at least 99% identity with positions 138-742 of SEQ ID NO: 982; and / or a VP3 protein containing an amino acid sequence having at least 99% identity with positions 203-742 of SEQ ID NO: 982. In some embodiments, the AAV capsid variant comprises a VP1 protein containing the amino acid sequence of SEQ ID NO: 982; a VP2 protein containing the amino acid sequence having positions 138-742 of SEQ ID NO: 982; and / or a VP3 protein containing the amino acid sequence having positions 203-742 of SEQ ID NO: 982.

[0024] In some embodiments, the viral genome of the AAV particle encodes the wild-type GBA1 protein. In some embodiments, the viral genome does not encode a hemagglutinin (HA) tag. In some embodiments, the viral genome encodes a human GBA1 protein, a canine GBA1 protein, or a horse GBA1 protein. In some embodiments, the viral genome encodes a human GBA1 protein. In some embodiments, the human GBA1 protein comprises the amino acid sequence of SEQ ID NO: 1775.

[0025] In some embodiments, the GBA1 coding sequence of the viral genome comprises a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 2002. In some embodiments, the GBA1 coding sequence comprises at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) of the same nucleotide sequence as SEQ ID NO: 2002. In some embodiments, the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002.

[0026] In some embodiments, the GBA1 coding sequence encodes a signal sequence comprising at least 90% of the same amino acid sequence as SEQ ID NO: 2005. In some embodiments, the signal sequence comprises the amino acid sequence of SEQ ID NO: 2005.

[0027] In some embodiments, the GBA1 coding sequence comprises the same amino acid sequence as SEQ ID NO: 2002.The nucleotide sequence of SEQ ID NO: 2001 is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical). In some embodiments, the GBA1 coding sequence comprises SEQ ID NO: 2001.

[0028] In some embodiments, the viral genome further comprises a miRNA (miR) binding site that regulates the expression of the encoded GBA1 protein in cells or tissues of the DRG, liver, heart, hematopoietic lineage, or combinations thereof.

[0029] In some embodiments, the viral genome further comprises a promoter effectively linked to the GBA1 coding sequence. In some embodiments, the promoter is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to the nucleotide sequence of SEQ ID NO: 1834. In some embodiments, the promoter comprises the nucleotide sequence of SEQ ID NO: 1834.

[0030] In some embodiments, the viral genome further comprises an enhancer. In some embodiments, the enhancer comprises a CMV Immediate Early (CMVie) enhancer. In some embodiments, the CMVie enhancer is at least 90% identical to the nucleotide sequence of SEQ ID NO: 1831 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical). In some embodiments, the CMVie enhancer comprises the nucleotide sequence of SEQ ID NO: 1831.

[0031] In some embodiments, the viral genome further comprises introns. In some embodiments, the intron is at least 90% identical to the nucleotide sequence of SEQ ID NO: 1842 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical). In some embodiments, the intron comprises the nucleotide sequence of SEQ ID NO: 1842.

[0032] In some embodiments, the viral genome further comprises a polyadenylated (polyadenylated) region. In some embodiments, this polyadenylated region is at least 90% identical to the nucleotide sequence of SEQ ID NO: 1846 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%).(or 100% identical). In some embodiments, the polyadenylated region comprises the nucleotide sequence of SEQ ID NO: 1846.

[0033] In some embodiments, the viral genome further comprises inverted terminal repeats (ITRs). In some embodiments, the ITR is at least 90% identical to the nucleotide sequence of SEQ ID NO: 1829 or SEQ ID NO: 1830 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical). In some embodiments, the ITR comprises the nucleotide sequence of SEQ ID NO: 1829 or SEQ ID NO: 1830. In some embodiments, the viral genome comprises a 5' ITR and a 3' ITR, wherein the 5' ITR comprises the nucleotide sequence of SEQ ID NO: 1829 and the 3' ITR comprises the nucleotide sequence of SEQ ID NO: 1830.

[0034] In some embodiments, the viral genome further includes one or more miR183 binding sites. In some embodiments, the viral genome includes four miR183 binding sites. In some embodiments, each of the four miR183 binding sites contains at least 70% identity with the nucleotide sequence of SEQ ID NO: 1847. In some embodiments, each of the four miR183 binding sites contains the nucleotide sequence of SEQ ID NO: 1847.

[0035] In some embodiments, the viral genome further includes a series of miR183 binding sites containing sequences that are at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the nucleotide sequence of SEQ ID NO: 1849. In some embodiments, the miR183 binding site series contains the nucleotide sequence of SEQ ID NO: 1849.

[0036] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR; (ii) a promoter; (iii) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; and (iv) a 3' ITR.

[0037] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR; (ii) a promoter; (iii) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the SEQ ID NO: 2002.The nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; and (iv) 3' ITR.

[0038] In some embodiments, the viral genome comprises: (i) a 5' ITR; (ii) a CMVie enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a chicken β-actin (CBA) promoter comprising the nucleotide sequence SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (iv) an intron comprising the nucleotide sequence SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) The GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; and (vi) 3' ITR.

[0039] In some embodiments, the viral genome comprises: (i) a 5' ITR; (ii) a CMVie enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a CBA promoter comprising the nucleotide sequence SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (iv) an intron comprising the nucleotide sequence SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; and (v) The GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001, and (vi) 3' ITR.

[0040] In some embodiments, the viral genome comprises: (i) a 5' ITR; (ii) a CMVie enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1831; (iii) a CBA promoter comprising the nucleotide sequence SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) an intron comprising the nucleotide sequence SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1842 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and (v) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence SEQ ID NO: 2002 or a nucleotide sequence identical to SEQ ID NO: 2002. 2002 is at least 90% identical (e.g., at least 93% identical) of the nucleotide sequence; (vi) a polyadenylated (polyadenylated) region containing the nucleotide sequence of SEQ ID NO: 1846 or at least 95% identical to the nucleotide sequence of SEQ ID NO: 1846; and (vii) 3' ITR.

[0041] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR; (ii) a CMVie enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a CBA promoter comprising the nucleotide sequence SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (iv) an intron comprising the nucleotide sequence SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) The GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleoside sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (vi) a polyadenylate region comprising SEQ ID NO:The nucleotide sequence of 1846 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1846; and (vii) 3' ITR.

[0042] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR containing the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) a CMVie enhancer containing the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a CBA promoter containing the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; and (iv) an intron containing the nucleotide sequence of SEQ ID NO: 1834. (v) A nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 2002; (vi) A polyadenylate region that contains a nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 1846; and (vii) A 3' ITR that contains a nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1830.

[0043] In some embodiments, the viral genome comprises: (i) a 5' ITR containing the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; and (ii) a CMVie enhancer containing the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831.(i) a nucleotide sequence; (iii) a CBA promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) a GBA1 coding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (vi) a polyadenylated region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 1834. 1846 is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) of a nucleotide sequence; and (vii) 3' ITR, which comprises the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) of SEQ ID NO: 1830.

[0044] In some embodiments, the viral genome comprises the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 97% identical) of SEQ ID NO: 2006. In some embodiments, the viral genome consists of the nucleotide sequence of SEQ ID NO: 2006.

[0045] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR; (ii) a promoter; (iii) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (iv) at least one miR183 binding site; and (v) a 3' ITR.

[0046] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR; (ii) a promoter; (iii) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (iv) at least one miR183 binding site; and (v) a 3' ITR.A nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (iv) at least one miR183 binding site; and (v) 3' ITR.

[0047] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) 5' ITR; (ii) a promoter; (iii) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (iv) at least one miR183 binding site sequence comprising at least one miR183 binding site and at least one spacer sequence; and (v) 3' ITR.

[0048] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR; (ii) a promoter; (iii) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (iv) at least one miR183 binding site sequence comprising at least one miR183 binding site and at least one spacer sequence; and (v) a 3' ITR.

[0049] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR; (ii) a CMVie enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a CBA promoter comprising the nucleotide sequence SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (iv) an intron comprising the nucleotide sequence SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) The GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (vi) at least one miR183 binding site; and (vii) a 3' ITR.

[0050] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR; (ii) a CMVie enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a CBA promoter comprising the nucleotide sequence SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (iv) an intron comprising the nucleotide sequence SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1842; and (v) a GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence SEQ ID NO: 2001 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1834. (vi) at least 90% identical (e.g., at least 94% identical) nucleotide sequence; (vii) at least one miR183 binding site; and (vii) 3' ITR.

[0051] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR; (ii) a CMVie enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a CBA promoter comprising the nucleotide sequence SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (iv) an intron comprising the nucleotide sequence SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) The GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (vi) a miR183 binding site series comprising the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical to SEQ ID NO: 1849; and (vii) a 3' ITR.

[0052] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR; (ii) a CMVie enhancer comprising a nucleotide sequence of SEQ ID NO: 1831 or at least 95% identical to SEQ ID NO: 1831 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iii) a CBA promoter comprising a nucleotide sequence of SEQ ID NO: 1834 or at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and (iv) an intron comprising a nucleotide sequence of SEQ ID NO: 1842 or at least 95% identical to SEQ ID NO: 1842 (e.g., at least 95%, at least 96%, at least 97%). (v) A nucleotide sequence that is at least 98% or at least 99% identical to; (vi) A GBA1 coding sequence that comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical to (e.g., at least 94% identical to) SEQ ID NO: 2001; (vi) A miR183 binding site series that comprises the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical to (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to) SEQ ID NO: 1849; and (vii) A 3' ITR.

[0053] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR containing the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) a CMVie enhancer containing the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a CBA promoter containing the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; and (iv) an intron containing the nucleotide sequence of SEQ ID NO: 1834.(v) A nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 2002; (vi) A GBA1 coding sequence comprising a nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (vi) At least one miR183 binding site comprising a nucleotide sequence of SEQ ID NO: 1847; (vii) A polyadenylate region comprising a nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1846; and (viii) A 3' ITR comprising a nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1846; 1830 has at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) nucleotide sequences.

[0054] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR containing the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) a CMVie enhancer containing the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a CBA promoter containing the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; and (iv) an intron containing the nucleotide sequence of SEQ ID NO: 1834. (v) A nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (vi) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises a nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (vi) At least one miR183 binding site comprising SEQ ID NO: 1842.(vii) A polyadenylated region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1846; and (viii) a 3' ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1830. Specification 9 / 390 pages 31 CN 121127597 A

[0055] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR containing the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) a CMVie enhancer containing the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a CBA promoter containing the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: (iv) A nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; (v) An intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (vi) A series of miR183 binding sites comprising the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; 1849 is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) of a nucleotide sequence; (vii) a polyadenylate region comprising the nucleotide sequence of SEQ ID NO: 1846 or identical to SEQ ID NO:1846 is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) of a nucleotide sequence; and (viii) 3' ITR, which contains the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) of SEQ ID NO: 1830.

[0056] In some embodiments, the viral genome comprises, in 5' to 3' order: (i) a 5' ITR containing the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) a CMVie enhancer containing the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) a CBA promoter containing the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834; and (iv) an intron containing the nucleotide sequence of SEQ ID NO: 1834. (v) A GBA1 coding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 2001; (vi) a miR183 binding site series comprising the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 1849; (vii) a polyadenylated region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) to SEQ ID NO: 1842; 1846 is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) of the nucleotide sequence; and (viii) 3' ITR, which contains the nucleotide sequence of SEQ ID NO: 1830 or is identical to SEQ ID NO: 1846.NO: 1830 has at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) nucleotide sequences.

[0057] In some embodiments, (i) the 5' ITR contains the nucleotide sequence of SEQ ID NO: 1829; (ii) the CMVie enhancer contains the nucleotide sequence of SEQ ID NO: 1831; (iii) the CBA promoter contains the nucleotide sequence of SEQ ID NO: 1834; (iv) the intron contains the nucleotide sequence of SEQ ID NO: 1842; (v) the GBA1 coding sequence contains the nucleotide sequence of SEQ ID NO: 2001; (vi) the miR183 binding site series contains the nucleotide sequence of SEQ ID NO: 1849; (vii) the polyadenylate region contains the nucleotide sequence of SEQ ID NO: 1846; and (viii) the 3' ITR contains the nucleotide sequence of SEQ ID NO: 1830.

[0058] In some embodiments, the viral genome comprises the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 97% identical) to SEQ ID NO: 2007. In some embodiments, the viral genome comprises the nucleotide sequence of SEQ ID NO: 2007. In some embodiments, the viral genome is composed of the nucleotide sequence of SEQ ID NO: 2007.

[0059] In some embodiments, this disclosure provides an AAV particle comprising: a viral genome comprising the nucleotide sequence of SEQ ID NO: 2001; an AAV capsid variant comprising a VP1 protein comprising the amino acid sequence of SEQ ID NO: 4; a VP2 protein comprising the amino acid sequence of SEQ ID NO: 4 at positions 138-742; and / or a VP3 protein comprising the amino acid sequence of SEQ ID NO: 4 at positions 203-742.

[0060] In some embodiments, this disclosure provides an AAV particle comprising: a viral genome comprising the nucleotide sequence of SEQ ID NO: 2001; an AAV capsid variant comprising a VP1 protein comprising the amino acid sequence of SEQ ID NO: 36; a VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 36; and / or a VP3 protein comprising the amino acid sequence of SEQ ID NO: 2001.The amino acid sequence at position 203-742 of SEQ ID NO: 36.

[0061] In some embodiments, this disclosure provides an AAV particle comprising: a viral genome containing the nucleotide sequence of SEQ ID NO: 2002; an AAV capsid variant containing a VP1 protein containing the amino acid sequence of SEQ ID NO: 4; a VP2 protein containing the amino acid sequence at position 138-742 of SEQ ID NO: 4; and / or a VP3 protein containing the amino acid sequence at position 203-742 of SEQ ID NO: 4.

[0062] In some embodiments, this disclosure provides an AAV particle comprising: a viral genome containing the nucleotide sequence of SEQ ID NO: 2002; an AAV capsid variant containing a VP1 protein containing the amino acid sequence of SEQ ID NO: 36; a VP2 protein containing the amino acid sequence at positions 138-742 of SEQ ID NO: 36; and / or a VP3 protein containing the amino acid sequence at positions 203-742 of SEQ ID NO: 36.

[0063] In some embodiments, this disclosure provides a cell comprising the AAV particle described herein. In some embodiments, the cell is a mammalian cell (e.g., HEK293 cell), an insect cell (e.g., Sf9 cell), or a bacterial cell.

[0064] In some embodiments, this disclosure provides a method for preparing the AAV particles described herein, the method comprising: providing a cell containing a viral genome, the viral genome containing a GBA1 coding sequence and a nucleic acid encoding the AAV capsid variant; and culturing the cell under conditions suitable for encapsulating the viral genome in the AAV capsid variant; thereby preparing the AAV particles.

[0065] In some embodiments, this disclosure provides a method for preparing AAV particles, wherein the viral genome of the AAV particles comprises (a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical to it (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); or (b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical to it (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and the AAV capsid variant of the AAV particles comprises: (i) a VP1 protein, which comprises SEQ ID NO: 2006.(i) an amino acid sequence of SEQ ID NO: 4 or an amino acid sequence having at least 90% identity with SEQ ID NO: 4 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity); (ii) a VP2 protein containing the amino acid sequence of SEQ ID NO: 4 at positions 138-742 or an amino acid sequence having at least 90% identity with SEQ ID NO: 4 at positions 138-742 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity); or (iii) a VP3 protein containing the amino acid sequence of SEQ ID NO: 4 at positions 203-742 or an amino acid sequence having at least 90% identity with SEQ ID NO: 4. Position 4, 203-742, has an amino acid sequence with at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity).

[0066] In some embodiments, this disclosure provides a method for preparing AAV particles, wherein the viral genome of the AAV particles comprises (a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical to it (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); or (b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical to it (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and the AAV capsid variant of the AAV particles comprises the amino acid sequence of SEQ ID NO: 4, the amino acid sequence at positions 138-742 of SEQ ID NO: 4, and / or the amino acid sequence at positions 203-742 of SEQ ID NO: 4.

[0067] In some embodiments, this disclosure provides a method for preparing AAV particles, wherein the viral genome of the AAV particles comprises: (a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical thereto (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); or (b) SEQ ID NO:The nucleotide sequence of SEQ ID NO: 36 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 36; and the AAV capsid variant of the AAV particle comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 36; and (ii) a VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 36 or an amino acid sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 36. The amino acid sequence at positions 138-742 of SEQ ID NO: 36 has at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity); and / or (iii) the VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 36 or having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) with the amino acid sequence at positions 203-742 of SEQ ID NO: 36.

[0068] In some embodiments, this disclosure provides a method for preparing AAV particles, wherein the viral genome of the AAV particles comprises: (a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical thereto (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical thereto); or (b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical thereto (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical thereto); and the AAV capsid variant of the AAV particles comprises the amino acid sequence of SEQ ID NO: 36, the amino acid sequence at positions 138-742 of SEQ ID NO: 36, and / or the amino acid sequence at positions 203-742 of SEQ ID NO: 36.

[0069] In some embodiments, this disclosure provides a method for preparing AAV particles, wherein the viral genome of the AAV particles comprises: (a) SEQ IDThe nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) therein; or (b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) therein; and the AAV capsid variant of the AAV particle comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) therein; (i) an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) with SEQ ID NO: 982; (ii) a VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) with SEQ ID NO: 982; and / or (iii) a VP3 protein comprising the amino acid sequence at positions 203 ... Positions 203-742 of 982 have an amino acid sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical).

[0070] In some embodiments, this disclosure provides a method for preparing AAV particles, wherein the viral genome of the AAV particles comprises: (a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); or (b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleus that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical).The AAV particle contains an amino acid sequence; and the AAV capsid variant of the AAV particle contains the amino acid sequence of SEQ ID NO: 982, the amino acid sequence at positions 138-742 of SEQ ID NO: 982, and / or the amino acid sequence at positions 203-742 of SEQ ID NO: 982.

[0071] In some embodiments, the method of preparing AAV particles further includes introducing a first nucleic acid molecule containing the viral genome into the cell prior to step (i). In some embodiments, the cell contains a second nucleic acid molecule encoding an AAV capsid variant. In some embodiments, the method of preparing AAV particles further includes introducing the second nucleic acid into the cell prior to step (i). In some embodiments, the cell contains mammalian cells (e.g., HEK293 cells), insect cells (e.g., Sf9 cells), or bacterial cells.

[0072] In some embodiments, this disclosure provides a pharmaceutical composition comprising the AAV particles described herein and pharmaceutically acceptable excipients.

[0073] In some embodiments, this disclosure provides a method of delivering AAV particles encoding the GBA1 protein to a subject, comprising administering to the subject an effective amount of the pharmaceutical composition or AAV particles described herein. In some embodiments, the subject has, has been diagnosed with, or is at risk of developing a GBA1-related condition. In some embodiments, the GBA1-related condition is a GBA1-related neurodegenerative or neuromuscular disease. In some embodiments, the GBA1-related condition is Parkinson's disease (PD), Parkinson's dementia (PDD), Gaucher disease (GD) (e.g., GD type 1, 2, or 3), Lewy body dementia (DLB), Lewy body dementia (LBD), multiple system atrophy (MSA), Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), isolated autonomic failure, brain iron deposition neurodegeneration type 1 (NBIA 1), or Hallewarden-Schpattz syndrome. In some embodiments, the subject has, has been diagnosed with, or is at risk of developing PD.

[0074] In some embodiments, this disclosure provides a method of treating a subject who has or has been diagnosed with a GBA1-related condition, comprising administering to the subject an effective amount of the pharmaceutical composition or AAV particles described herein. In some embodiments, the GBA1-related condition is a GBA1-related neurodegenerative or neuromuscular disorder. In some embodiments, the GBA1-related condition is PD, PDD, GD (e.g., GD type 1, 2, or 3), DLB, LBD, MSA, AD, ALS, isolated autonomic failure, NBIA 1, or Hallewarden-Schpattz syndrome. In some embodiments, the GBA1-related neurodegenerative or neuromuscular disorder is...Neuromuscular degeneration or neurodegenerative disease is PD. In some embodiments, GBA1-related neuromuscular degeneration or neurodegenerative disease is GD (e.g., GD type 1, 2, or 3). In some embodiments, GBA1-related neuromuscular degeneration or neurodegenerative disease is GD type 1. In some embodiments, GBA1-related neuromuscular degeneration or neurodegenerative disease is GD type 2. In some embodiments, GBA1-related neuromuscular degeneration or neurodegenerative disease is GD type 3. In some embodiments, GBA1-related neuromuscular degeneration or neurodegenerative disease is DLB. In some embodiments, GBA1-related neuromuscular degeneration or neurodegenerative disease is LBD.

[0075] In some embodiments, this disclosure provides a method of treating a subject with PD or diagnosed with PD, comprising administering to the subject an effective amount of the pharmaceutical composition or AAV particles described herein.

[0076] In some embodiments, this disclosure provides a method of treating a subject with GD (e.g., GD type 1, 2, or 3) or diagnosed with GD (e.g., GD type 1, 2, or 3), comprising administering to the subject an effective amount of the pharmaceutical composition or AAV particles described herein. In some embodiments, the GD is GD type 1. In some embodiments, the GD is GD type 2. In some embodiments, the GD is GD type 3.

[0077] In some embodiments, this disclosure provides a method of treating a subject with LBD, comprising administering to the subject an effective amount of the pharmaceutical composition or AAV particles described herein. In some embodiments, this disclosure provides a method of treating a subject with DLB, comprising administering to the subject an effective amount of the pharmaceutical composition or AAV particles described herein.

[0078] In some embodiments, this disclosure provides a method of treating a subject wherein the subject has one or more mutations in the GBA1 gene. In some embodiments, the subject has lower GC enzyme activity compared to the GC enzyme activity of a subject without GBA1-related conditions. In some embodiments, the level of GC enzyme activity is measured by a 4-MUG assay or a SensoLyte blue glucocerebrosidase assay.

[0079] In some embodiments, the treatment halts the progression of the condition in the subject. In some embodiments, the treatment improves the condition. In some embodiments, the treatment alters one or more biomarkers of the condition. In some embodiments, the one or more biomarkers include GC enzyme activity, the content of glucocerebrosidase and other glycolipids (e.g., in immune cells such as macrophages), and the content of synuclein aggregates (e.g., Lewy bodies).The amount, content of neurofilament light chains, or combinations thereof. In some embodiments, the treatment improves at least one symptom of the condition. In some embodiments, the at least one symptom includes developmental delay, progressive encephalopathy, progressive dementia, ataxia, myoclonus, oculomotor dysfunction, bulbar palsy, general weakness, limb tremor, depression, visual hallucinations, cognitive decline, or combinations thereof.

[0080] In some embodiments, the subject is a human.

[0081] In some embodiments, the AAV particles are delivered to cells, tissues, or regions of the CNS, such as regions of the brain or spinal cord, such as the parenchyma, cortex, substantia nigra, caudate nucleus, cerebellum, striatum, corpus callosum, cerebellum, brainstem, caudate nucleus-putamen, thalamus, superior colliculus, spinal cord, or combinations thereof, wherein the AAV particles or a pharmaceutical composition containing the AAV particles are delivered via intravenous administration.

[0082] In some embodiments, this disclosure provides a method of treating a subject, further comprising assessing, for example, measuring the subject's GBA1 expression level in cells, tissues, or body fluids, such as GBA1 gene expression level, GBA1 mRNA expression level, and / or GBA1 protein expression level. In some embodiments, the GBA1 protein expression level is measured by ELISA, Western blot, or immunohistochemistry. In some embodiments, the assessment of GBA1 expression level is performed before and after administration of AAV. In some embodiments, the GBA1 expression level before administration is compared with the GBA1 expression level after administration.

[0083] In some embodiments, the method of delivery or treatment includes assessing the GBA1 expression level in cells or tissues of the central nervous system (e.g., parenchyma). In some embodiments, the GBA1 protein expression level of the subject increases after administration relative to the subject's GBA1 protein expression level before administration.

[0084] In some embodiments, the method further includes assessing, for example, measuring the level of GC enzyme activity in the subject, as per the specification 14 / 390 page 36 CN 121127597 A.

[0085] In some embodiments, administration of the AAV particles or pharmaceutical composition described herein causes an increase in: (i) GC enzyme activity in the subject's cells, tissues (e.g., cells or tissues of the CNS, such as the cortex, striatum, thalamus, cerebellum, and / or brainstem) and / or body fluids (e.g., CSF and / or serum), optionally wherein the GC enzyme activity in the subject increases by at least 2-fold, at least 3-fold, at least 4-fold, or at least 5-fold after administration relative to the GC enzyme activity in the subject prior to administration; (ii) GC enzyme activity in the subject's CNS tissues (e.g., cortex, striatum, thalamus, cerebellum, brainstem, and / or spinal cord).The content of viral genome (VG) per cell in the subject, optionally, wherein the content of VG per cell in the subject's CNS tissue is increased by more than 50 VG per cell relative to the content of VG per cell in the subject's peripheral tissue; and / or (iii) the expression of GBA1 mRNA in the subject's cells or tissues (e.g., cells or tissues of the CNS, such as the cortex, thalamus, and / or brainstem), optionally, wherein the expression of GBA1 mRNA in the subject is increased by at least 100-1300 times after administration compared to the expression of GBA1 mRNA in the subject before administration.

[0086] In some embodiments, this disclosure provides a method of treating a subject, which further includes administering to the subject an additional agent suitable for treating or preventing GBA1-related conditions. In some implementations, the additional agent comprises enzyme replacement therapy (ERT) (e.g., imiglucerase, velaglucerase α, or taliglucerase α); substrate reduction therapy (SRT) (e.g., eliglustat or miglustat), levodopa, carbidopa, safinamide, dopamine agonists (e.g., quetiapine, clozapine, pramipexole, rotigotine, or ropinirole), or anticholinergics (e.g., benztropine). Or trihexyphenidyl), cholinesterase inhibitors (e.g., rivastigmine, donepezil, or galantamine), N-methyl-d-aspartate (NMDA) receptor antagonists (e.g., memantine), or combinations thereof. In some embodiments, the method further includes administering a blood transfusion to the subject. In some embodiments, the method further includes administering an immunosuppressant to the subject. In some embodiments, the immunosuppressant comprises a corticosteroid (e.g., prednisone, prednisolone, methylprednisolone, and / or dexamethasone), rapamycin, mycophenolate mofetil, tacrolimus, rituximab.

[0087] In some embodiments, this disclosure provides the pharmaceutical compositions or AAV particles described herein for the treatment of GBA1-related conditions. In some embodiments, the GBA1-related condition is PD, PDD, GD (e.g., GD type 1, 2, or 3), DLB, LBD, MSA, AD, ALS, isolated autonomic failure, NBIA 1, or Hallewarden-Spattz syndrome. In some embodiments, the GBA1-related condition is PD. In some embodiments, the GBA1-related condition is GD (e.g., GD type 1, 2, or 3). In some embodiments, GD is GD type 1. In some embodiments, GD is GD type 2. In some embodiments, GD is GD type 3. In some embodiments, the GBA1-related condition is DLB. In some embodiments, the GBA1-related condition is LBD.

[0088] In some embodiments, this disclosure provides the use of the pharmaceutical composition or AAV particles described herein in the preparation of a medicament for treating GBA1-related conditions. In some embodiments, the GBA1-related condition is PD, PDD, GD (e.g., GD type 1, 2, or 3), DLB, LBD, MSA, AD, ALS, isolated autonomic failure, NBIA 1, or Hallewarden-Schpattz syndrome. In some embodiments, the GBA1-related condition is PD. In some embodiments, the GBA1-related condition is GD (e.g., GD type 1, 2, or 3). In some embodiments, GD is GD type 1. In some embodiments, GD is GD type 2. In some embodiments, GD is GD type 3. In some embodiments, the GBA1-related condition is DLB. In some embodiments, the GBA1-related condition is LBD.

[0089] The listed embodiments

[0090] 1. An adeno-associated virus (AAV) particle comprising an AAV capsid variant (e.g., an AAV9 capsid variant) and a viral genome comprising a β-glucocerebrosidase 1 (GBA1) coding sequence (e.g., encoding human GBA1 protein), wherein the AAV capsid variant comprises 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 an amino acid sequence of SPH; and (iii)[N3] comprises X4, X5, and X6, wherein at least one of X4, X5, or X6 is a basic amino acid, such as K or R.

[0091] 2. The AAV particle of embodiment 1, wherein X4, X5, or both of [N3] are K.

[0092] 3. The AAV particle of embodiment 1 or 2, wherein X4, X5, or X6 of [N3] are R.

[0093] 4. An AAV particle as described in any one of embodiments 1 to 3, wherein: (a) X4 of [N3] is: K, S, A, V, T, G, F, W, V, N, or R; (b) X5 of [N3] is: S, K, T, F, I, L, Y, H, M, or R; and / or (c) X6 of [N3] is: G, A, R, M, I, N, T, Y, D, P, V, L, E, W, N, Q, K, or S; Optionally, the AAV capsid variant comprises an amino acid modification of any of the aforementioned amino acids in (a)-(c), for example, a conservative substitution.

[0094] 5. AAV particles as described in any one of embodiments 1 to 4, wherein [N3] includes 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.

[0095] 6. An AAV particle as described in any one of embodiments 1 to 5, wherein [N3] is or includes 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.

[0096] 7. AAV particles as described in any one of embodiments 1 to 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), SPHAS (SEQ ID NO: 4708), SPHKS (SEQ ID NO: 4709), SPHAS (SEQ ID NO: 4709), SPHK ...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).

[0097] 8. AAV particles as described in any one of embodiments 1 to 7, wherein [N2]-[N3] are or comprise: (i) 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: 959), SPHKKN (SEQ ID NO: 950), SPHKKN (SEQ ID NO: 954), SPHKVRM (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: 959), SPHKKN (SEQ ID NO: 950), SPHKKN ...0), SPHKKN (SEQ ID NO: 950), SPHKKN (SEQ 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:(ii) any part of the amino acid sequence in (i), such as any 2, 3, 4 or 5 amino acids therein, such as an amino acid sequence of consecutive amino acids; (iii) an amino acid sequence containing one, two or three but not more than four modified amino acids relative to any of the amino acid sequences in (i); or (iv) an amino acid sequence containing one, two or three but not more than four different amino acids relative to any of the amino acid sequences in (i).

[0098] 9. An AAV particle as described in any one of embodiments 1 to 8, wherein the AAV capsid variant comprises an amino acid other than G (e.g., V, R, D, E, M, T, I, S, A, N, L, K, H, P, W, or C) at position 453 according to any one of SEQ ID NO: 36-59, 138, 981, or 982, an amino acid other than S (e.g., V, L, N, D, H, R, P, G, T, I, A, E, Y, M, or Q) at position 454, and / or an amino acid other than G (e.g., C, L, D, E, Y, H, V, A, N, P, or S) at position 455.

[0099] 10. An AAV particle of any one of embodiments 1 to 8, wherein the AAV capsid variant comprises amino acid G at position 453, amino acid S at position 454, and amino acid G at position 455, as specified in SEQ ID NO: 138 or 981.

[0100] 11. An AAV particle of any one of embodiments 1 to 9, wherein the AAV capsid variant comprises amino acid G at position 453, amino acid H at position 454, and amino acid D at position 455, as specified in SEQ ID NO: 138 or 982.

[0101] 12. An AAV particle of any one of embodiments 1 to 11, wherein [N1] comprises X1, X2, and X3, wherein at least one of X1, X2, or X3 is G.

[0102] 13. An AAV particle as described in any one of embodiments 1 to 12, wherein: (a) X1 of [N1] is: G, V, R, D, E, M, T, I, S, A, N, L, K, H, P, W, or C; (b) X2 of [N1] is: S, V, L, N, D, H, R, P, G, T, I, A, E, Y, M, or Q; and / or (c) X3 of [N1] is: G, C, L, D, E, Y, H, V, A, N, P, or S; Optionally, the AAV capsid variant comprises an amino acid modification of any of the aforementioned amino acids in (a)-(c), for example, a conservative substitution. Specification 17 / 390 pages 39 CN 121127597 A

[0103] 14.AAV particles as described in any of embodiments 1 to 13, wherein [N1] includes 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.

[0104] 15. An AAV particle as described in any one of embodiments 1 to 14, wherein [N1] is or includes 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.

[0105] 16. The AAV particles of any one of embodiments 1 to 15, wherein [N1]-[N2] comprise: (i) 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), ... 4765), TGSPH (SEQ ID NO: 4766), SVSPH (SEQ ID NO: 4767), IGSPH (SEQ ID NO: 4768), DGSPH (SEQ ID NO:(ii) any portion of the amino acid sequence in (i), such as any 2, 3, or 4 amino acids, such as a sequence of consecutive amino acids; (iii) any part of the amino acid sequence in (i), such as any 2, 3, or 4 amino acids therein; (i) A sequence containing one, two, or three, but no more than four modified amino acid sequences relative to any of the amino acid sequences in (i); or (iv) A sequence containing one, two, or three, but no more than four different amino acids relative to any of the amino acid sequences in (i).

[0106] 17. AAV particles as described in any one of embodiments 1 to 16, wherein [N1]-[N2] are or comprise: (i) 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); (ii)(i) any part of the amino acid sequence in (i), such as any 2, 3, 4 or 5 amino acids therein, such as an amino acid sequence of consecutive amino acids; (iii) an amino acid sequence containing one, two or three but no more than four modified amino acids relative to any of the amino acid sequences in (i); or (iv) an amino acid sequence containing one, two or three but no more than four different amino acids relative to any of the amino acid sequences in (i).

[0107] 18. AAV particles as described in any one of embodiments 1 to 17, wherein [N1]-[N2]-[N3] comprises: (i) 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: 4850), RGSPHSK (SEQ ID NO: 4851), RGSPHSK (SEQ ID NO: 4849), SGSPHAS (SEQ ID NO: 4840), RGSPHAS ...42), SGSPHKS (SEQ ID NO: 4843), SGSPHKS (SEQ ID NO: 4844), SGSPHAS (SEQ ID NO: 4845), RGSPHAS (SEQ ID NO: 4851), RGSPHSK (SEQ ID NO: 4849), SGSPHAS (SEQ ID NO: 4840), RGSPHAS (SEQ ID NO: 4845), RGSPHAS (SEQ ID NO: 4845), RGSPHAS ( 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), SSGSPHTK (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: (ii) any part of the amino acid sequence in (i), such as any 2, 3, 4, 5 or 6 amino acids therein, for example...Ming Shu 19 / 390 page 41 CN 121127597 A such amino acid sequence of consecutive amino acids; (iii) an amino acid sequence containing one, two or three but not more than four modified amino acids relative to any of the amino acid sequences in (i); or (iv) an amino acid sequence containing one, two or three but not more than four different amino acids relative to any of the amino acid sequences in (i).

[0108] 19. AAV particles as described in any one of embodiments 1 to 18, wherein [N1]-[N2]-[N3] are or comprise: (i) 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), GSGSPHSKA (SEQ ID NO: 4917), GSGSPHKS (SEQ ID NO: 4918), GSGSPHSKA (SEQ ID NO: 4919), GSGSPHKS (SEQ ID NO: 4919 ... NO: 4916), CSHSPHKSG (SEQ ID NO: 4917), GQSSPHRSG (SEQ ID NO: 4918), GRGSPHASR (SEQ ID NO: 4919), GRGSPHSKA (SEQ ID NO: 4920), GGSSPHKFG (SEQ ID NO: 4921), GGSSPHKIG (SEQ ID NO: 4922), GGSSPHKLG (SEQ ID NO: 4923), GGSSPHKTS (SEQ ID NO: 4924), GSGSPHKTT (SEQ ID NO: 4925), GGSSPHKTY (SEQ ID NO: 4926), GSGSPHKYG (SEQ ID NO: 4927), GSGSPHSKD (SEQ ID NO: 4928), GGSSPHSKP (SEQ ID NO: 4929), GGSSPHTRG (SEQ ID NO: 4930), GGSPHSVRG (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 (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) GSSCSPHRKA (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: (SEQ ID NO: 4961).(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: 说明书 20 / 390 页 42 CN 121127597 A 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: It should be noted that there seems to be some incomplete or incorrect information in the original text, especially the part "说明书 20 / 390 页 42 CN 121 A 4991" which is not in a standard format. You may want to double-check and correct the original text for a more accurate translation.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 (ii) any portion of the amino acid sequence in (i), such as any 2, 3, 4, 5, 6, 7, or 8 amino acids therein, such as a sequence of consecutive amino acids; (iii) Relative to any of the amino acid sequences in (i), there is an amino acid sequence comprising one, two, or three, but not more than four modified amino acid sequences; or (iv) relative to any of the amino acid sequences in (i), there is an amino acid sequence comprising one, two, or three, but not more than four different amino acids.

[0109] 20. An AAV particle as described in any of embodiments 1 to 19, wherein [N3] comprises SK, KA, KS, or SG.

[0110] 21.21. An AAV particle as described in any one of embodiments 1 to 20, wherein [N3] is or contains SKA, KSG, or KYG.

[0111] 22. An AAV particle as described in any one of embodiments 1 to 21, wherein [N2]-[N3] contains SPHSK (SEQ ID NO: 4701), SPHKS (SEQ ID NO: 4704), or SPHKY (SEQ ID NO: 4715).

[0112] 23. An AAV particle as described in any one of embodiments 1 to 22, wherein [N2]-[N3] is or contains SPHSKA (SEQ ID NO: 941).

[0113] 24. An AAV particle as described in any one of embodiments 1 to 22, wherein [N2]-[N3] is or contains SPHKSG (SEQ ID NO: 946).

[0114] 25. An AAV particle as described in any one of embodiments 1 to 22, wherein [N2]-[N3] is or contains SPHKYG (SEQ ID NO: 966).

[0115] 26. An AAV particle as described in any one of embodiments 1 to 25, wherein [N1] contains GS, SG, GH, or HD.

[0116] 27. An AAV particle as described in any one of embodiments 1 to 26, wherein [N1] is or contains GSG.

[0117] 28. An AAV particle as described in any one of embodiments 1 to 26, wherein [N1] is or contains GHD.

[0118] 29. An AAV particle as described in any one of embodiments 1 to 23 or 26 to 27, wherein [N1]-[N2]-[N3] contains SGSPHSK (SEQ ID NO: 4839).

[0119] 30. An AAV particle as described in any one of embodiments 1 to 22, 24, 26 or 28, wherein [N1]-[N2]-[N3] comprises HDSPHKS (SEQ ID NO: 4840).

[0120] 31. An AAV particle as described in any one of embodiments 1 to 22 or 25 to 27, wherein [N1]-[N2]-[N3] comprises SGSPHKYG (SEQ ID NO: 5027). Specification 21 / 390 pages 43 CN 121127597 A

[0121] 32. An AAV particle as described in any one of embodiments 1 to 8, 10, 12 to 23, 26 to 27 or 29, wherein [N1]-[N2]-[N3] is or comprises GGSPHSKA (SEQ ID NO: 4697).

[0122] 33. An AAV particle as described in any one of embodiments 1 to 9, 11 to 22, 24, 26, 28 or 30, wherein [N1]-[N2]-[N3] is or contains GHDSPHKSG (SEQ ID NO:4698).

[0123] 34. AAV particles as described in any one of embodiments 1 to 8, 10, 12 to 22, 25 to 27 or 31, wherein [N1]-[N2]-[N3] are or contain GGSPHKYG (SEQ ID NO: 4927).

[0124] 35. AAV particles as described in any one of embodiments 1 to 34, wherein [N1]-[N2]-[N3] replace positions 453-455 according to SEQ ID NO: 138.

[0125] 36. An AAV particle as described in any one of embodiments 1 to 35, wherein the AAV capsid variant 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).

[0126] 37. An AAV particle as described in any one of embodiments 1 to 36, wherein the AAV capsid variant comprises an amino acid other than Q (e.g., W, K, R, G, L, V, S, P, H, K, I, M, A, E, or F) at position 462 numbered according to SEQ ID NO: 981, 982, 36, 37, 39, 40, 42-46, 48, 49, 50, 52, 53, 56, or 57, 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 (e.g., Y, C, K, T, H, R, D, V, S, P, G, W, E, F, A, I, M, Q, or L) at position 463, an amino acid other than Q (e.g., G, K, H, R, T, L, D, A, P, I, F, V, M, W, Y, S, E, N, or Y) at position 464, and / or an amino acid other than Q (e.g., H, K, R, T, L, D, A, P, I, F, V, M, W, Y, S, E, N, or Y) at position 465, and / or an amino acid other than Q (e.g., H, K, R, T, L, D, A, P, I, F, V, M, W, Y, S, E, N, or Y) at position 465. L, R, W, K, A, P, E, M, I, S, G, N, Y, C, V, T, D, or V).

[0127] 38. An AAV particle as described in any one of embodiments 1 to 37, wherein the AAV capsid variant comprises: (a) amino acid Q at position 456, amino acid N at position 457, amino acid Q at position 458, and / or amino acid Q at position 459 according to SEQ ID NO: 138; or (b) according to SEQ ID NO:Amino acid Q at position 462, amino acid N at position 463, amino acid Q at position 464, and / or amino acid Q at position 465, numbered 981, 982, 36, 37, 39, 40, 42-46, 48, 49, 50, 52, 53, 56 or 57.

[0128] 39. An AAV particle as described in any one of embodiments 1 to 38, wherein the AAV capsid variant further comprises [N4], wherein [N4] comprises X7 X8 X9 X10, and wherein: (a) X7 is: Q, W, K, R, G, L, V, S, P, H, K, I, M, A, E or F; (b) X8 is: N, Y, C, K, T, H, R, D, V, S, P, G, W, E, F, A, I, M, Q or L; (c) 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) X10 is: Q, H, L, R, W, K, A, P, E, M, I, S, G, N, Y, C, V, T, D or V; Optionally, the AAV capsid variant comprises an amino acid modification of any of the aforementioned amino acids in (a)-(d), for example, a conservative substitution.

[0129] 40. The AAV particle of embodiment 39, wherein: (a) X7 of [N4] is Q or R; (b) X8 of [N4] is N or R; (c) X9 of [N4] is Q or R; and (d) X10 of [N4] is Q, L, or R.

[0130] 41. AAV particles as described in embodiment 39 or 40, wherein [N4] is or comprises: Specification 22 / 390 pages 44 CN 121127597 A (i) 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: 5040), 5041), QRKL (SEQ ID NO: 5042), LRRQ (SEQ ID NO: 5043), QRLR (SEQ ID NO: 5044), QRRL (SEQID 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 DESCRIPTION 23 / 390 PAGES 45 CN 121127597 A (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 IDNO: 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: (ii) any portion of the amino acid sequence in (i), such as any two or three amino acids therein, such as a sequence of consecutive amino acids; (iii) a sequence containing one, two, or three, but no more than four modified amino acid sequences relative to any one of the amino acid sequences in (i); or (iv) a sequence containing one, two, or three, but no more than four modified amino acid sequences. Relative to any of the amino acid sequences in (i), an amino acid sequence comprising one, two, or three but no more than four different amino acids.

[0131] 42. An AAV particle as described in any of embodiments 39 to 41, wherein [N1]-[N2]-[N3]-[N4] is or comprises: (i) SEQ ID NO: 1800-1808, 1832, 1833, 1835-1841, 1843-1845, 1848, 1851,The amino acid sequence of any one of 1852, 1854-2000, 2003, 2004, 2008-2241, 6421-6441, 6443-6456; Specification 24 / 390 pages 46 CN 121127597 A (ii) comprising any part of the amino acid sequence in (i), such as any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 amino acids, such as a sequence of consecutive amino acids; (iii) relative to any one of the amino acid sequences in (i), comprising one, two or three but not more than four modified amino acid sequences; or (iv) relative to any one of the amino acid sequences in (i), comprising one, two or three but not more than four different amino acids.

[0132] 43. An AAV particle as described in any one of embodiments 39 to 42, wherein [N1]-[N2]-[N3]-[N4] is or contains GSGSPHSKAQNQQ (SEQ ID NO: 1801).

[0133] 44. An AAV particle as described in any one of embodiments 39 to 42, wherein [N1]-[N2]-[N3]-[N4] is or contains GHDSPHKSGQNQQ (SEQ ID NO: 1800).

[0134] 45. An AAV particle as described in any one of embodiments 39 to 42, wherein [N1]-[N2]-[N3]-[N4] is or contains GSGSPHKYGQNQQT (SEQ ID NO: 910).

[0135] 46. An AAV particle as described in any one of embodiments 1 to 45, wherein the AAV capsid variant comprises an amino acid other than T (e.g., S, Y, M, A, C, I, R, L, D, F, V, Q, N, H, E, or G) at position 450, an amino acid other than I (e.g., M, P, E, N, D, S, A, T, G, Q, F, V, L, C, H, R, W, or L) at position 451, and / or an amino acid other than N (e.g., M, E, G, Y, W, T, I, Q, F, V, A, L, I, P, K, R, H, S, D, or S) at position 452.

[0136] 47. An AAV particle as described in any one of embodiments 1 to 46, wherein the AAV capsid variant comprises amino acid T at position 450, amino acid I at position 451, and / or amino acid N at position 452, according to any one of SEQ ID NO: 138, 981, or 982.

[0137] 48. An AAV particle as described in any one of embodiments 1 to 47, wherein the AAV capsid variant further comprises [NO], wherein [NO] comprises XAXB and XC, wherein: (a) XA is: T, S, Y, M, A, C, I, R, L, D, F, V, Q, N, H, E or G; (b) XB is: I, M, P, E, N, D, S, A, T, G, Q, F, V, L, C, H, R, W or L; and (c) 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 of any of the aforementioned amino acids in (a)-(c), such as a conservative substitution. 49. The AAV particle of embodiment 48, wherein [NO] is or includes 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.

[0139] 50. AAV particles as described in embodiment 48 or 49, wherein [N0]-[N1]-[N2]-[N3]-[N4] are or comprise: (i) an amino acid sequence of any one of SEQ ID NO: 2242-2886; (ii) any portion of the amino acid sequence in (i), such as any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, ...Page 47 of document 25 / 390 CN 121127597 A 12, 13, 14 or 15 amino acids, for example, an amino acid sequence of consecutive amino acids; (iii) a sequence of one, two or three but not more than four modified amino acids relative to any of the amino acid sequences in (i); or (iv) a sequence of one, two or three but not more than four different amino acids relative to any of the amino acid sequences in (i).

[0140] 51. An AAV particle as described in any of embodiments 48 to 50, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or contains TINGSGSPHSKAQNQQ (SEQ ID NO: 2242).

[0141] 52. An AAV particle as described in any one of embodiments 48 to 50, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or contains TINGHDSPHKSGQNQQ (SEQ ID NO: 2243).

[0142] 53. An AAV particle as described in any one of embodiments 48 to 52, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or contains TINGSGSPHKYGQNQQT (SEQ ID NO: 5246).

[0143] 54. An AAV particle as described in any one of embodiments 1 to 53, wherein [N1]-[N2]-[N3] are present in ring IV.

[0144] 55. An AAV particle as described in any one of embodiments 48 to 54, wherein [N0] and [N4] are present in ring IV.

[0145] 56. An AAV particle as described in any one of embodiments 48 to 55, wherein [NO] is present immediately after position 449 according to SEQ ID NO: 138.

[0146] 57. An AAV particle as described in any one of embodiments 48 to 56, wherein [NO] is present immediately after position 449 according to any one of SEQ ID NO: 36-59, 981, or 982.

[0147] 58. An AAV particle as described in any one of embodiments 48 to 57, wherein [NO] replaces positions 450, 451, and 452 according to SEQ ID NO: 138 (e.g., T450, I451, and N452).

[0148] 59. An AAV particle as described in any one of embodiments 48 to 58, wherein [NO] replaces positions 450-452 (e.g., T450, I451, and N452) according to any one of the numbers in SEQ ID NO: 36-59, 981, or 982.

[0149] 60. An AAV particle as described in any one of embodiments 48 to 59, wherein [NO] corresponds to SEQ ID NO: 36-59,Positions 450-452 of any one of 138, 981, or 982.

[0150] 61. An AAV particle as described in any one of embodiments 48 to 60, wherein [NO] is present immediately after position 449, and wherein [NO] replaces positions 450-452 (e.g., T450, I451, and N452), which are numbered according to SEQ ID NO: 138.

[0151] 62. An AAV particle as described in any one of embodiments 48 to 61, wherein [NO] is present immediately after position 449, and wherein [NO] replaces positions 450-452 (e.g., T450, I451, and N452), which are numbered according to any one of SEQ ID NO: 36-59, 981, or 982.

[0152] 63. An AAV particle as described in any one of embodiments 1 to 62, wherein [N1] is present immediately after position 452 of the amino acid sequence number according to SEQ ID NO: 138.

[0153] 64. An AAV particle as described in any one of embodiments 1 to 63, wherein [N1] is present immediately after position 452 of the sequence number according to SEQ ID NO: 981 or 982.

[0154] 65. An AAV particle as described in any one of embodiments 1 to 61, wherein [N1] replaces positions 453-455 of the sequence number according to SEQ ID NO: 138 (e.g., G453, S454, and G455).

[0155] 66. An AAV particle as described in any one of embodiments 1 to 64, wherein [N1] replaces position 453 of the sequence number according to SEQ ID NO: 138 (e.g., G453).

[0156] 67. An AAV particle as described in any one of embodiments 1 to 65, wherein [N1] replaces positions 453-455 (e.g., G453, S454, and G455) numbered according to SEQ ID NO: 981. Specification 26 / 390 pages 48 CN 121127597 A

[0157] 68. An AAV particle as described in any one of embodiments 1 to 65 or 67, wherein [N1] replaces positions 453-455 numbered according to SEQ ID NO: 982.

[0158] 69. An AAV particle as described in any one of embodiments 1 to 65, 67, or 68, wherein [N1] is present immediately after position 452, and wherein [N1] replaces positions 453-455 (e.g., G453, S454, and G455) numbered according to SEQ ID NO: 138.

[0159] 70. An AAV particle as described in any one of embodiments 1 to 64 or 66, wherein [N1] is present immediately after position 452, and wherein [N1] replaces position 453 (e.g., G453), these positions according to SEQ IDNO: 138.

[0160] 71. An AAV particle as described in any of embodiments 1 to 64, 66 or 70, wherein [N1] is present immediately after position 452, and wherein [N1] replaces positions 453-455, which are numbered according to SEQ ID NO: 4, 36, 981 or 982.

[0161] 72. An AAV particle as described in any of embodiments 1 to 71, wherein [N1] corresponds to positions 453-455 numbered according to any of SEQ ID NO: 4, 36-59, 981 or 982.

[0162] 73. An AAV particle as described in any of embodiments 1 to 72, wherein the AAV capsid variant comprises an amino acid other than S at position 454 numbered according to SEQ ID NO: 138, 981 or 982 and / or an amino acid other than G at position 455.

[0163] 74. An AAV particle of any one of embodiments 1 to 73, wherein the AAV capsid variant comprises amino acid H at position 454 and amino acid D at position 455, as specified in SEQ ID NO: 138 or 982.

[0164] 75. An AAV particle of any one of embodiments 1 to 74, wherein the AAV capsid variant comprises a substitution (e.g., S454H) at position 454 and / or a substitution (e.g., G455D) at position 455, as specified in SEQ ID NO: 138.

[0165] 76. An AAV particle of any one of embodiments 1 to 75, wherein the AAV capsid variant comprises amino acid H at position 454 and amino acid D at position 455, and further comprises the amino acid sequence SPHSKA (SEQ ID NO: 941) immediately following position 455, these positions being specified in SEQ ID NO: 138.

[0166] 77. An AAV particle of any one of embodiments 1 to 76, wherein the AAV capsid variant comprises amino acid H at position 454 and amino acid D at position 455 according to SEQ ID NO: 982.

[0167] 78. An AAV particle of any one of embodiments 1 to 77, wherein the AAV capsid variant comprises amino acid H at position 454 and amino acid D at position 455, and further comprises the amino acid sequence SPHSKA (SEQ ID NO: 941) immediately following position 455, these positions being numbered according to SEQ ID NO: 982.

[0168] 79. An AAV particle of any one of embodiments 1 to 72, wherein the AAV capsid variant comprises amino acid S at position 454 and amino acid G at position 455 according to SEQ ID NO: 138.

[0169] 80.AAV particle of any one of embodiments 1 to 72 or 79, wherein the AAV capsid variant comprises amino acid S at position 454 and amino acid G at position 455, and further comprises the amino acid sequence SPHSKA (SEQ ID NO: 941) immediately following position 455, these positions being numbered according to SEQ ID NO: 138.

[0170] 81. AAV particle of any one of embodiments 1 to 72, 79 or 80, wherein the AAV capsid variant comprises amino acid S at position 454 and amino acid G at position 455, numbered according to SEQ ID NO: 981.

[0171] 82. AAV particle of any one of embodiments 1 to 72 or 79 to 81, wherein the AAV capsid variant comprises amino acid S at position 454 and amino acid G at position 455, and further comprises the amino acid sequence SPHSKA (SEQ ID NO: 941) immediately following position 455, these positions being numbered according to SEQ ID NO: 981.

[0172] 83. An AAV particle as described in any one of embodiments 1 to 82, wherein [N2] is present immediately after position 455 according to the number SEQ ID NO: 138.

[0173] 84. An AAV particle as described in any one of embodiments 1 to 83, wherein [N2] corresponds to positions 456-458 of SEQ ID NO: 981 or CN 121127597 A 982 on page 27 / 390 of the specification (e.g., S456, P457, H458).

[0174] 85. An AAV particle as described in any one of embodiments 1 to 83, wherein [N2] corresponds to positions 456-458 of SEQ ID NO: 4 or any one of 36-59 (e.g., S456, P457, H458).

[0175] 86. An AAV particle as described in any one of embodiments 1 to 85, wherein [N2]-[N3] are present immediately after position 455 according to SEQ ID NO: 138.

[0176] 87. An AAV particle as described in any one of embodiments 1 to 86, wherein [N2] is present immediately after position 455 according to SEQ ID NO: 4, 36, 981 or 982.

[0177] 88. An AAV particle as described in any one of embodiments 1 to 87, wherein [N2]-[N3] are present immediately after position 455 according to SEQ ID NO: 4, 36, 981 or 982.

[0178] 89. An AAV particle as described in any one of embodiments 1 to 88, wherein [N2]-[N3] correspond to positions 456-461 of SEQ ID NO: 981.(e.g., S456, P457, H458, S459, K460, A461).

[0179] 90. An AAV particle as described in any one of embodiments 1 to 88, wherein [N2]-[N3] correspond to positions 456-461 of SEQ ID NO: 982 (e.g., S456, P457, H458, K459, S460, G461).

[0180] 91. An AAV particle as described in any one of embodiments 1 to 90, wherein [N2] is present immediately after [N1].

[0181] 92. An AAV particle as described in any one of embodiments 1 to 64, 66, 70 or 71, wherein [N3] is present immediately after [N2] and replaces positions 454 and 455 (e.g., S454 and G455), which are numbered according to SEQ ID NO: 138.

[0182] 93. An AAV particle as described in any one of embodiments 1 to 64, 66, 70, 71 or 92, wherein [N3] is present immediately after [N1]-[N2] and replaces positions 454 and 455 (e.g., S454 and G455), which are numbered according to SEQ ID NO: 138.

[0183] 94. An AAV particle as described in any one of embodiments 39 to 93, wherein [N4] is present immediately after position 455 numbered according to SEQ ID NO: 138.

[0184] 95. An AAV particle as described in any one of embodiments 39 to 94, wherein [N4] replaces positions 456-459 numbered according to SEQ ID NO: 138 (e.g., Q456, N457, Q458 and Q459).

[0185] 96. An AAV particle as described in any one of embodiments 39 to 95, wherein [N4] corresponds to positions 462-465 of SEQ ID NO: 4, 36, 981 or 982 (e.g., Q462, N463, Q464, Q465).

[0186] 97. An AAV particle as described in any one of embodiments 39 to 96, wherein [N2]-[N3]-[N4] replace positions 456-459 (e.g., Q456, N457, Q458 and Q459) numbered according to SEQ ID NO: 138.

[0187] 98. An AAV particle as described in any of embodiments 39 to 97, wherein [N2]-[N3]-[N4] is present immediately after position 455, and wherein [N2]-[N3]-[N4] replaces positions 456-459 (e.g., Q456, N457, Q458, and Q459), these positions being numbered according to SEQ ID NO: 138.

[0188] 99. An AAV particle as described in any of embodiments 39 to 98, wherein [N2]-[N3]-[N4] corresponds to SEQ ID NO:Positions 456-465 of SEQ ID NO: 981 (e.g., S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465).

[0189] 100. AAV particles as described in any of embodiments 39 to 98, wherein [N2]-[N3]-[N4] correspond to positions 456-465 of SEQ ID NO: 982 (e.g., S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465).

[0190] 101. AAV particles as described in any of embodiments 39 to 98, wherein [N2]-[N3]-[N4] correspond to positions 456-465 of SEQ ID NO: 4 or any of 36-59.

[0191] 102. AAV particles as described in any of embodiments 39 to 101, wherein [N1]-[N2]-[N3]-[N4] replace positions 453-459 of the root specification page 28 / 390, CN 121127597 A according to SEQ ID NO: 138 (e.g., G453, S454, G455, Q456, N457, Q458 and Q459).

[0192] 103. An AAV particle as described in any of embodiments 39 to 102, wherein [N1]-[N2]-[N3]-[N4] is present immediately after position 452, and wherein [N1]-[N2]-[N3]-[N4] replaces positions 453-459 (e.g., G453, S454, G455, Q456, N457, Q458 and Q459), which are numbered according to SEQ ID NO: 138.

[0193] 104. AAV particles as described in any one of embodiments 39 to 99, 102 or 103, wherein [N1]-[N2]-[N3]-[N4] correspond to positions 453-465 of SEQ ID NO: 981 (e.g., G453, S454, G455, S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465).

[0194] 105. AAV particles as described in any one of embodiments 39 to 98, 100, 102 or 103, wherein [N1]-[N2]-[N3]-[N4] correspond to positions 453-465 of SEQ ID NO: 982 (e.g., G453, H454, D455, S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465).

[0195] 106.AAV particles as described in any of embodiments 39 to 98, 102 or 103, wherein [N1]-[N2]-[N3]-[N4] correspond to positions 453-465 of SEQ ID NO: 4 or any of 36-59.

[0196] 107. AAV particles as described in any of embodiments 1 to 99 or 102 to 104, wherein [N1]-[N2]-[N3] correspond to positions 453-461 of SEQ ID NO: 981 (e.g., G453, S454, G455, S456, P457, H458, S459, K460, A461).

[0197] 108. AAV particles as described in any one of embodiments 1 to 98, 100, 102, 103 or 105, wherein [N1]-[N2]-[N3] correspond to positions 453-461 of SEQ ID NO: 982 (e.g., G453, H454, D455, S456, P457, H458, K459, S460, G461).

[0198] 109. AAV particles as described in any one of embodiments 39 to 98, 102, 103 or 106, wherein [N1]-[N2]-[N3] correspond to positions 453-461 of any one of SEQ ID NO: 36-59.

[0199] 110. AAV particles as described in any of embodiments 48 to 109, wherein [N0]-[N1]-[N2]-[N3]-[N4] replace positions 450-459 (e.g., T450, I451, N452, G453, S454, G455, Q456, N457, Q458 and Q459) as numbered according to SEQ ID NO: 138.

[0200] 111. An AAV particle as described in any of embodiments 48 to 110, wherein [N0]-[N1]-[N2]-[N3]-[N4] is present immediately after position 449, and wherein [N0]-[N1]-[N2]-[N3]-[N4] replaces positions 450-459 (e.g., T450, I451, N452, G453, S454, G455, Q456, N457, Q458 and Q459), these positions being numbered according to SEQ ID NO: 138.

[0201] 112. AAV particles as described in any one of embodiments 48 to 99, 102 to 104 or 106, wherein [N0]-[N1]-[N2]-[N3]-[N4] correspond to positions 450-465 of SEQ ID NO: 981 (e.g., T450, I451, N452, G453, S454, G455, S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465).

[0202] 113. AAV particles as described in any one of embodiments 48 to 98, 100, 102, 103, 105 or 108, wherein [N0]-[N1]-[N2]-[N3]-[N4] correspond to positions 450-465 of SEQ ID NO: 982 (e.g., T450, I451, N452, G453, H454, D455, S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465).

[0203] 114. AAV particles as described in any one of embodiments 48 to 98, 102, 103, 106 or 109, wherein [N0]-[N1]-[N2]-[N3]-[N4] correspond to positions 450-465 of any one of SEQ ID NO: 36-59.

[0204] 115. AAV particles as described in any one of embodiments 39 to 114, wherein [N4] replaces positions 462-465 (e.g., Q462, N463, Q464 and Q465) numbered according to SEQ ID NO: 4, 36, 981 or 982.

[0205] 116. An AAV particle as described in any of embodiments 39 to 115, wherein [N2]-[N3]-[N4] replaces positions 462-465 (e.g., Q462, N463, Q464, and Q465) numbered according to SEQ ID NO: 4, 36, 981, or 982. Specification 29 / 390 pages 51 CN 121127597 A

[0206] 117. An AAV particle as described in any of embodiments 39 to 116, wherein [N2]-[N3]-[N4] is present immediately after position 455, and wherein [N2]-[N3]-[N4] replaces positions 462-465 (e.g., Q462, N463, Q464, and Q465), which are numbered according to SEQ ID NO: 4, 36, 981, or 982.

[0207] 118. An AAV particle as described in any one of embodiments 1 to 117, wherein [N3] is present immediately following [N2].

[0208] 119. An AAV particle as described in any one of embodiments 1 to 118, wherein the AAV capsid variant comprises [N2]-[N3] from the N-terminus to the C-terminus.

[0209] 120. An AAV particle as described in any one of embodiments 1 to 119, wherein the AAV capsid variant comprises [N1]-[N2]-[N3] from the N-terminus to the C-terminus.

[0210] 121. An AAV particle as described in any one of embodiments 48 to 120, wherein the AAV capsid variant comprises [NO]-[N1]-[N2]-[N3] from the N-terminus to the C-terminus.

[0211] 122.123. An AAV particle as described in any of embodiments 39 to 121, wherein the AAV capsid variant comprises [N1]-[N2]-[N3]-[N4] from the N-terminus to the C-terminus.

[0212] 124. An AAV particle as described in any of embodiments 48 to 122, wherein the AAV capsid variant comprises [NO]-[N1]-[N2]-[N3]-[N4] from the N-terminus to the C-terminus.

[0213] 125. An AAV particle as described in any of the preceding embodiments, wherein the AAV capsid variant contains an amino acid other than T at position 460 according to SEQ ID NO: 138 (e.g., N, I, C, H, R, L, D, Y, A, M, Q, I, E, K, P, G, or S).

[0214] 125. An AAV particle as described in any of the foregoing embodiments, wherein the AAV capsid variant comprises an amino acid N, I, C, H, R, L, D, Y, A, M, Q, I, E, K, P, G, or S at position 460 according to SEQ ID NO: 138.

[0215] 126. An AAV particle as described in any of the foregoing embodiments, wherein the AAV capsid variant comprises an amino acid other than T (e.g., N, I, C, H, R, L, D, Y, A, M, Q, I, E, K, P, G, or S) at position 466 according to any of SEQ ID NO: 36-59, 981, or 982.

[0216] 127. An AAV particle as described in any of the foregoing embodiments, wherein the AAV capsid variant comprises amino acid N, I, C, H, R, L, D, Y, A, M, Q, I, E, K, P, G, or S at position 466 according to any of the numbers in SEQ ID NO: 36-59, 981, or 982.

[0217] 128. An AAV particle as described in any of the foregoing embodiments, wherein the AAV capsid variant comprises amino acid other than K (e.g., E, N, or T) at position 449 according to any of the numbers in SEQ ID NO: 36-59, 138, 981, or 982.

[0218] 129. An AAV particle as described in any of the foregoing embodiments, wherein the AAV capsid variant comprises amino acid E, N, or T at position 449 according to any of the numbers in SEQ ID NO: 36-59, 138, 981, or 982.

[0219] 130. An adeno-associated virus (AAV) particle comprising an AAV capsid variant (e.g., an AAV9 capsid variant) and a viral genome comprising a β-glucocerebrosidase 1 (GBA1) coding sequence (e.g., encoding a human GBA1 protein), wherein the AAV capsid variant comprises [A][B] (SEQ ID NO: 4694), wherein: (i)[A] contains the amino acid sequence GSGSPH (SEQ ID NO: 4695); and (ii) [B] contains X1 X2 X3 X4 X5 X6 X7, wherein: (a) X1 is: S, C, F or V; (b) X2 is: K, L, R, I, E, Y, V or S; (c) X3 is: A, R, L, G, I, Y, S, F or W; (d) X4 is: W, Q, R, G, L, V, S or F; Specification 30 / 390 pages 52 CN 121127597 A (e) X5 is: N, Y, R, C, K or L; (f) X6 is: Q, G, K, R, T, L or Y; and (g) X7 is: Q, L, R or V; Optionally, the AAV capsid variant contains an amino acid modification, such as a conservative substitution, of any of the aforementioned amino acids in (a)-(g).

[0220] 131. AAV particles as described in embodiment 130, wherein

[0221] (a) X1 is S; (b) X2 is K or L; (c) X3 is A, R or L; (d) X4 is Q or R; (e) X5 is N or R; (f) X6 is Q or R; and (g) X7 is Q, L or R.

[0222] 132. AAV particles as described in embodiment 130 or 131, wherein [B] comprises: (i) SLLWNQQ (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: 5259), CLQNQQ (SEQ ID NO: 5250), SLRRNQQ (SEQ ID NO: 5251), SLRRNQQ (SEQ ID NO: 5252), SLRRNQQ (SEQ ID NO: 5253), SLRRNQQ (SEQ ID NO: 5254), SLRRNQQ (SEQ ID NO: 5255), SLRRNQQ (SEQ ID NO: 5256), SLRRNQQ (SEQ ID NO: 5257), SLRRNQQ (SEQ ID NO: 5258), SKAFRLQ (SEQ ID NO: 5259), CLQNQQ (SEQ ID NO: 5259), SLRRNQQ (SEQ ID NO: 5250), SLRRNQQ (SEQ ID NO: 52 ... 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 (ii) any part of the amino acid sequence in (i), such as any 2, 3, 4, 5 or 6 amino acids, such as an amino acid sequence of consecutive amino acids; (iii) an amino acid sequence containing one, two or three but not more than four modified amino acids relative to any of the amino acid sequences in (i); or (iv) an amino acid sequence containing one, two or three but not more than four different amino acids relative to any of the amino acid sequences in (i).

[0223] 133. AAV particles as described in any one of embodiments 130 to 132, wherein [A][B] comprises: (i) 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: 5285), GSGSPHSRGRNQQ (SEQ ID NO: 2066), GSGSPHSEIVNQQ (SEQ ID NO: 5286), GSGSPHSLLWNQQ (SEQ ID NO: 5286), GSGSPHSLRRN ...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); (ii) any portion of the amino acid sequence in (i), such as any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11Or 12 amino acids, such as an amino acid sequence of consecutive amino acids; (iii) relative to any of the amino acid sequences in (i), comprising one, two, or three but not more than four modified amino acid sequences; or (iv) relative to any of the amino acid sequences in (i), comprising one, two, or three but not more than four different amino acid sequences.

[0224] 134. An AAV particle as described in any of embodiments 130 to 133, wherein the AAV capsid variant further comprises one, two, or all of the amino acid other than T (e.g., S, Y, or G) at position 450, the amino acid other than I (e.g., M or L) at position 451, and / or the amino acid other than N (e.g., S) at position 452 according to SEQ ID NO: 138.

[0225] 135. An AAV particle as described in any of embodiments 130 to 134, wherein the AAV capsid variant further comprises S at position 450 and M at position 451 according to SEQ ID NO: 138.

[0226] 136. An AAV particle as described in any one of embodiments 130 to 134, wherein the AAV capsid variant further comprises Y at position 450, L at position 451, and S at position 452 according to SEQ ID NO: 138.

[0227] 137. An AAV particle as described in any one of embodiments 130 to 134, wherein the AAV capsid variant further comprises G at position 450, L at position 451, and S at position 452 according to SEQ ID NO: 138.

[0228] 138. An AAV particle as described in any one of embodiments 130 to 137, wherein [A][B] is present in ring IV.

[0229] 139. An AAV particle as described in any one of embodiments 130 to 138, wherein [A] is present immediately after position 452 according to SEQ ID NO: 138.

[0230] 140. An AAV particle as described in any one of embodiments 130 to 139, wherein [A] replaces positions 453-455 (e.g., G453, S454, G455) numbered according to SEQ ID NO: 138.

[0231] 141. An AAV particle as described in any one of embodiments 130 to 140, wherein [A] is present immediately after position 452, and wherein [A] replaces positions 453-455 (e.g., G453, S454, G455) numbered according to SEQ ID NO: 138.

[0232] 142. An AAV particle as described in any one of embodiments 130 to 141, wherein [B] is present immediately after [A].

[0233] 143.AAV particles as described in any of embodiments 130 to 142, wherein [B] replaces positions 456-459 (e.g., Q456, N457, Q458, Q459) according to SEQ ID NO: 138. Specification 32 / 390 pages 54 CN 121127597 A

[0234] 144. AAV particles as described in any of embodiments 130 to 143, wherein [A][B] replaces positions 453-459 (e.g., G453, S454, G455, Q456, N457, Q458, Q459) according to SEQ ID NO: 138.

[0235] 145. An AAV particle as described in any of embodiments 130 to 144, wherein [A][B] is present immediately after position 452, and wherein [A][B] replaces positions 453-459 (e.g., G453, S454, G455, Q456, N457, Q458, Q459), which are numbered according to SEQ ID NO: 138.

[0236] 146. An AAV particle as described in any of embodiments 130 to 145, wherein the AAV capsid variant comprises [A][B] from the N-terminus to the C-terminus.

[0237] 147. An adeno-associated virus (AAV) particle comprising an AAV capsid variant and a viral genome comprising a β-glucocerebrosidase 1 (GBA1) coding sequence (e.g., encoding human GBA1 protein), wherein the AAV capsid variant comprises [A][B] (SEQ ID NO: 4699), wherein: (i) [A] comprises X1 X2 X3 X4 X5 X6, wherein (a) X1 is T, M, A, C, I, R, L, D, F, V, Q, N or H; (b) X2 is I, P, E, N, D, S, A, T, M or Q; (c) X3 is N, E, G, Y, W, M, T, I, K, Q, F, S, V, A or L; (d) X4 is G, D, R or E; (e) X5 is H, Q, N or D; (f) X6 is D or R; Optionally, the AAV capsid variant comprises an amino acid modification of any of the aforementioned amino acids in (a)-(f), for example, a conservative substitution; and (ii) [B] comprises SPHKSG (SEQ ID NO: 946).

[0238] 148. The AAV particle of embodiment 147, wherein

[0239] (a) X1 is: T, M, A, or I; (b) X2 is: E, I, or D; (c) X3 is: N, Q, Y, I, M, or V; (d) X4 is G; (e) X5 is H; and (f) X6 is D.

[0240] 149. The AAV particle of embodiment 147 or 148, wherein [A] comprises: (i)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 SPECIFICATION Page 33 / 390 55 CN 121127597 A 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 IDNO: 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: 5342) 5349), QQQGHD (SEQ ID NO: 5350), TPQDHD (SEQ ID NO: 5351), HDWGHD (SEQ (ii) any part of the amino acid sequence in (i), such as any 2, 3, 4 or 5 amino acids therein, such as an amino acid sequence of consecutive amino acids; (iii) an amino acid sequence containing one, two or three but not more than four modified amino acids relative to any of the amino acid sequences in (i); or (iv) an amino acid sequence containing one, two or three but not more than four different amino acids relative to any of the amino acid sequences in (i).

[0241] 150. An AAV particle as described in any one of embodiments 147 to 149, wherein [A][B] comprises: (i) TINGHDSPHKS (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), EGGHDSPHKS (SEQ ID NO: 5365 ...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), NNMNGHDSPHKS (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), Instructions 34 / 390 Page 56 CN 121127597 A IEGGHDSPHKS (SEQ ID NO: 5411) (ii) comprising any part of the amino acid sequence in (i), such as any 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids therein, such as an amino acid sequence of consecutive amino acids; (iii) comprising one, two or three but not more than four modified amino acid sequences relative to any of the amino acid sequences in (i); or (iv) comprising one, two or three but not more than four different amino acid sequences relative to any of the amino acid sequences in (i).

[0242] 151. An AAV particle as described in any one of embodiments 147 to 150, wherein the AAV capsid variant further comprises one, two, three, four, or all of the following amino acids: at position 456 other than Q (e.g., R or L), at position 457 other than N (e.g., H, K, or R), at position 458 other than Q (e.g., R or T), at position 459 other than Q (H), and / or at position 460 other than T (N or S).

[0243] 152. An AAV particle as described in any one of embodiments 147 to 151, wherein the AAV capsid variant further comprises R at position 456 as described in SEQ ID NO: 138.

[0244] 153. An AAV particle as described in any one of embodiments 147 to 151, wherein the AAV capsid variant further comprises one, two, three, four, or all of the following amino acids as described in SEQ ID NO: 138.L at position 456 of SEQ ID NO: 138.

[0245] 154. An AAV particle as described in any one of embodiments 147 to 153, wherein the AAV capsid variant further comprises H at position 457 and R at position 458 according to SEQ ID NO: 138.

[0246] 155. An AAV particle as described in any one of embodiments 147 to 153, wherein the AAV capsid variant further comprises K at position 457 and N at position 460 according to SEQ ID NO: 138.

[0247] 156. An AAV particle as described in any one of embodiments 147 to 153, wherein the AAV capsid variant further comprises T at position 458, H at position 459 and S at position 460 according to SEQ ID NO: 138.

[0248] 157. An AAV particle as described in any one of embodiments 147 to 151, wherein the AAV capsid variant further comprises R at position 456, R at position 457, and R at position 458 according to SEQ ID NO: 138.

[0249] 158. An AAV particle as described in any one of embodiments 147 to 157, wherein [A][B] is present in ring IV.

[0250] 159. An AAV particle as described in any one of embodiments 147 to 158, wherein [A] is present immediately after position 449 according to SEQ ID NO: 138.

[0251] 160. An AAV particle as described in any one of embodiments 147 to 159, wherein [A] replaces positions 450-453 according to SEQ ID NO: 138 (e.g., T450, I451, N452, G453).

[0252] 161. An AAV particle as described in any of embodiments 147 to 160, wherein [A] is present immediately after position 449, and wherein [A] replaces positions 450-453 (e.g., T450, I451, N452, G453), which are numbered according to SEQ ID NO: 138.

[0253] 162. An AAV particle as described in any of embodiments 147 to 161, wherein [A][B] replaces positions 450-455 (e.g., T450, I451, N452, G453, S454, G455) numbered according to SEQ ID NO: 138.

[0254] 163. An AAV particle as described in any of embodiments 147 to 162, wherein [A][B] is present immediately after position 449, and wherein [A][B] replaces positions 450-455 (e.g., T450, I451, N452, G453, S454, G455), which are numbered according to SEQ ID NO: 138.

[0255] 164.AAV particles of any one of embodiments 147 to 163, wherein [B] is present immediately after [A] and replaces positions 454 and 455 (e.g., S454 and G455), which are numbered according to SEQ ID NO: 138. Specification 35 / 390 pages 57 CN 121127597 A

[0256] 165. AAV particles of any one of embodiments 147 to 164, wherein [B] is present immediately after position 455 numbered according to SEQ ID NO: 4, 36, 981 or 982.

[0257] 166. AAV particles of any one of embodiments 147 to 165, wherein [B] is present immediately after [A].

[0258] 167. AAV particles of any one of embodiments 147 to 166, wherein the AAV capsid variant comprises [A][B] from the N-terminus to the C-terminus.

[0259] 168. An adeno-associated virus (AAV) particle comprising an AAV capsid variant (e.g., an AAV9 capsid variant) and a viral genome comprising a β-glucocerebrosidase 1 (GBA1) coding sequence (e.g., encoding a human GBA1 protein), wherein the AAV capsid variant comprises an amino acid sequence having the following formula: [N1]-[N2]-[N3] (SEQ ID NO: 6407), wherein: (i) [N1] comprises X1, X2, and X3, wherein X2 is S and X3 is G; (ii) [N2] comprises the amino acid sequence SPH; and (iii) [N3] comprises X4, X5, and X6, wherein X5 is K.

[0260] 169. An AAV particle as described in 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, such as a conservative substitution, of any of the aforementioned amino acids in (i) or (ii).

[0261] 170. An AAV particle as described in embodiment 168 or 169, wherein X4 is S and / or X5 is A.

[0262] 171. An AAV particle as described in any one of embodiments 168 to 170, wherein [N3] comprises SK, TK, NK, AK, KA, KV, KT, KS, KG, KR, KL, or KN.

[0263] 172. An AAV particle as described in any one of embodiments 168 to 171, wherein [N3] is or includes SKA, SKV, SKT, SKS, SKG, SKR, TKA, NKA, SKL, SKN, or AKA.

[0264] 173. An AAV particle as described in any one of embodiments 168 to 172, wherein [N3] is or includes SKA.

[0265] 174.AAV particles as described in any of embodiments 168 to 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).

[0266] 175. An AAV particle as described in any one of embodiments 168 to 174, wherein [N2]-[N3] is or comprises: (i) SPHSKA (SEQ ID NO: 941), SPHSKV (SEQ ID NO: 4737), SPHSKT (SEQ ID NO: 4731), SPHSKS (SEQ ID NO: 962), SPHSKG (SEQ ID NO: 4732), SPHSKR (SEQ ID NO: 978), SPHTKA (SEQ ID NO: 4739), SPHNKA (SEQ ID NO: 4734), SPHSKL (SEQ ID NO: 960), SPHSKN (SEQ ID NO: 4735), or SPHAKA (SEQ ID NO: 4736); (ii) comprising any portion of the amino acid sequence in (i), such as any 2, 3, 4, or 5 amino acids therein, such as a sequence of consecutive amino acids; (iii) Relative to any of the amino acid sequences in (i), there is a modified amino acid sequence comprising one, two, or three but no more than four modified amino acid sequences; or (iv) relative to any of the amino acid sequences in (i), there is a modified amino acid sequence comprising one, two, or three but no more than four different amino acids.

[0267] 176. An AAV particle as described in any of embodiments 168 to 175, wherein [N2]-[N3] is or comprises SPHSKA (SEQ ID NO: 941). Specification 36 / 390 pages 58 CN 121127597 A

[0268] 177. An AAV particle as described in any of embodiments 168 to 176, wherein the AAV capsid variant comprises an amino acid other than G (e.g., M, T, I, E, S, A, N, V, L, K, H, P, R, W, or D) at position 453 according to SEQ ID NO: 138 or 981.

[0269] 178. An AAV particle as described in any one of embodiments 168 to 177, wherein the AAV capsid variant comprises amino acid G at position 453 according to SEQ ID NO: 138 or 981.

[0270] 179. An AAV particle as described in any one of embodiments 168 to 178, wherein X1 of [N1] is 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 of any of the aforementioned amino acids, for example, a conserved substitution.

[0271] 180. An AAV particle as described in any of embodiments 168 to 179, wherein [N1] comprises SG, GS, MS, TS, IS, ES, SS, AS, NS, VS, LS, KS, HS, PS, RS, WS, or DS.

[0272] 181. An AAV particle as described in any of embodiments 168 to 180, wherein [N1] is or comprises: GSG, MSG, TSG, ISG, ESG, SSG, ASG, NSG, VSG, LSG, KSG, HSG, PSG, RSG, WSG, or DSG.

[0273] 182. An AAV particle as described in any of embodiments 168 to 181, wherein [N1] is or comprises GSG.

[0274] 183. AAV particles as described in any one of embodiments 168 to 182, wherein [N1]-[N2] comprises SGSPH (SEQ ID NO: 4752).

[0275] 184. AAV particles as described in any one of embodiments 168 to 183, wherein [N1]-[N2] are or comprise: (i) GSGSPH (SEQ ID NO: 4695), MSGSPH (SEQ ID NO: 4798), TSGSPH (SEQ ID NO: 4800), ISGSPH (SEQ ID NO: 4801), ESGSPH (SEQ ID NO: 4803), SSGSPH (SEQ ID NO: 4804), ASGSPH (SEQ ID NO: 4806), NSGSPH (SEQ ID NO: 4807), VSGSPH (SEQ ID NO: 4786), LSGSPH (SEQ ID NO: 4808), KSGSPH (SEQ ID NO: 4810), HSGSPH (SEQ ID NO: 4811), PSGSPH (SEQ ID NO: 4813), RSGSPH (SEQ ID NO: 4815), WSGSPH (ii) any part of the amino acid sequence in (i), such as any 2, 3, 4 or 5 amino acids therein, such as an amino acid sequence of consecutive amino acids; (iii) a sequence of one, two or three but not more than four modified amino acids relative to any one of the amino acid sequences in (i); or (iv) a sequence of one, two or three but not more than four modified amino acids relative to any one of the amino acid sequences in (i).Amino acid sequences of different amino acids.

[0276] 185. AAV particles as described in any one of embodiments 168 to 184, wherein [N1]-[N2]-[N3] are or comprise: (i) GSGSPHSKA (SEQ ID NO: 4697), GSGSPHSKV (SEQ ID NO: 4956), MSGSPHSKA (SEQ ID NO: 4957), TSGSPHSKA (SEQ ID NO: 4959), ISGSPHSKA (SEQ ID NO: 4960), GSGSPHSKT (SEQ ID NO: 4962), ESGSPHSKA (SEQ ID NO: 4963), SSGSPHSKA (SEQ ID NO: 4964), GSGSPHSKS (SEQ ID NO: 4953), ASGSPHSKA (SEQ ID NO: 4966), NSGSPHSKA (SEQ ID NO: 4967), VSGSPHSKA (SEQ ID NO: 4967), VSGSPHSKA (SEQ ID NO: 4968), NSGSPHSKA (SEQ ID NO: 4967), VSGSPHSKA (SEQ ID NO: 4968), NSGSPHSKA (SEQ ID NO: 4969), VSGSPHSKA (SEQ ID NO: 4969 ... 4913), LSGSPHSKA (SEQ ID NO: 4968), KSGSPHSKA (SEQ ID NO: 4970), GSGSPHSKG (SEQ ID NO: 4972), GSGSPHSKR (SEQ ID NO: 4945), HSGSPHSKA (SEQ ID NO: 4973), PSGSPHSKA (SEQ ID NO: 4975), RGSSPHSKA (SEQ ID NO: 4977), GGSSPHTKA (SEQ ID NO: 4978), WSGSPHSKA (SEQ ID NO: 4980), DSGSPHSKA (SEQ ID NO: 4981), GSGSPHNKA (SEQ ID NO: 4983), GSGSPHSKL (SEQ ID NO: 4943), GSGSPHSKN (SEQ ID NO: 4994) or GSGSPHAKA (SEQ ID NO: 4995); Instructions 37 / 390 Page 59 CN 121127597 A (ii) Contains any portion of the amino acid sequence in (i), such as any 2, 3, 4, 5, 6, 7, 8, or 9 amino acids therein, such as an amino acid sequence of consecutive amino acids; (iii) Contains one, two, or three, but not more than four modified amino acid sequences relative to any of the amino acid sequences in (i); or (iv) Contains one, two, or three, but not more than four modified amino acid sequences relative to any of the amino acid sequences in (i).Amino acid sequences of different amino acids.

[0277] 186. AAV particles as described in any one of embodiments 168 to 185, wherein [N1]-[N2]-[N3] are or contain GGSPHSKA (SEQ ID NO: 4697).

[0278] 187. An AAV capsid variant of any one of embodiments 168 to 186, comprising an amino acid other than Q (e.g., R, P, H, L, K, I, G, S, M, or E) at position 456 according to SEQ ID NO: 138, an amino acid other than N (e.g., D, V, S, P, T, G, Y, W, E, R, H, K, F, A, I, L, or M) at position 457, an amino acid other than N (e.g., D, V, S, P, T, G, Y, W, E, R, H, K, F, A, I, L, or M) at position 458, an amino acid other than Q (e.g., R, L, A, P, H, T, I, F, K, V, M, G, W, Y, S, E, N, or D) at position 459, an amino acid other than Q (e.g., H, K, A, L, P, E, M, I, S, N, R, Y, C, V, T, W, D, G) at position 460, and / or an amino acid other than T (e.g., ... I, N, S, H, R, L, D, Y, A, or Q).

[0279] 188. An AAV particle as described in any one of embodiments 168 to 187, wherein the AAV capsid variant comprises according to SEQ ID NO: Amino acids other than Q (e.g., R, P, H, L, K, I, G, S, M, or E) at position 462 of number 981, amino acids other than N (e.g., D, V, S, P, T, G, Y, W, E, R, H, K, F, A, I, L, or M) at position 464, amino acids other than Q (e.g., R, L, A, P, H, T, I, F, K, V, M, G, W, Y, S, E, N, or D) at position 465, amino acids other than Q (e.g., H, K, A, L, P, E, M, I, S, N, R, Y, C, V, T, W, D, G) at position 466, and / or amino acids other than T (e.g., I, N, S, H, R, L, D, Y, A, or Q) at position 466.

[0280] 189. An AAV particle as described in any one of embodiments 168 to 188, wherein the AAV capsid variant comprises amino acid Q at position 456, amino acid N at position 457, amino acid Q at position 458, amino acid Q at position 459, and / or amino acid T at position 460, as specified in SEQ ID NO: 138.

[0281] 190. An AAV particle as described in any one of embodiments 168 to 189, wherein the AAV capsid variant comprises amino acid Q at position 462, amino acid N at position 463, amino acid Q at position 464, amino acid Q at position 465, and / or amino acid T at position 466, as specified in SEQ ID NO: 981.

[0282] 191. An AAV particle as described in any one of embodiments 168 to 190, wherein the AAV capsid variant further comprises [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, the AAV capsid variant includes an amino acid modification of any of the aforementioned amino acids from (a) to (e), such as a conservative substitution.

[0283] 192. The AAV particle of embodiment 191, wherein [N4] is or comprises: (i) QNQQT (SEQ ID NO: 5412), QNRHT (SEQ ID NO: 5413), RDQQT (SEQ ID NO: 5414), PNLQT (SEQ ID NO: 5415), HVRQT (SEQ ID NO: 5416), PNQHT (SEQ ID NO: 5417), QSQQT (SEQ ID NO: 5418), QNQQI (SEQ ID NO: 5419), QPAKT (SEQ ID NO: 5420), QTQQN (SEQ ID NO: 5421), QNLAT (SEQ ID NO: 5422), QNQLT (SEQ ID NO: 5423), QGQQT (SEQ ID NO: 5424), QNQQT (SEQ ID NO: 5425), QNQQT (SEQ ID NO: 5426), QNQQT (SEQ ID NO: 5427), QNQQT (SEQ ID NO: 5428), QNQQT (SEQ ID NO: 5429), QNQQT (SEQ ID NO: 5420), QNQQT (SEQ ID NO: 5421), QNQQT (SEQ ID NO: 5422), QNQLT (SEQ ID NO: 5423), QGQQT (SEQ ID NO: 5424), QNQQT (SEQ ID NO: 5425), QNQQT (SEQ ID NO: 5426), QNQQT (SEQ ID NO: 5427), QNQQT (SEQ ID NO: 5428), QNQQT (SEQ ID NO: 5429), QNQQT (SEQ ID NO: 5421), QNQQT (SEQ ID NO: 5422), QNQQT (SEQ ID NO: 5423), QNQQT (SEQ ID NO: 5424 5424), LNRQS (SEQ ID NO: 5425), HNQQT (SEQ ID NO: 5426), QNPPT (SEQ ID NO: 5427), QNLQT (SEQ ID NO: 5428), QYQQT (SEQ ID NO: 5429), QDQET (SEQ ID NO: 5430), QNHQT (SEQ ID NO: 5431), QDQQT (SEQ ID NO: 5432), HWQQT (SEQ ID NO: 5433), PNQQT (SEQ ID NO: 5434), QNQLI (SEQ ID NO: 5435), PEQQT (SEQ IDNO: 5436)、QRTMT (SEQ ID NO: 5437)、QNQQH (SEQ ID NO: 5438)、LHQHT (SEQ ID NO: 5439)、QHRIT (SEQ ID NO: 5440)、QYIHT (SEQ ID NO: 5441)、QKFET (SEQ ID NO: 5442)、QFPST (SEQ ID NO: 5443)、HNQQR (SEQ ID NO: 5443) ID NO: 5444)、QAIKT (SEQ ID NO: 5445)、QNRQT (SEQ ID NO: 5446)、QYQHT (SEQ ID NO: 5447)、QNPQS (SEQ ID NO: 5448)、QHQLT (SEQ ID NO: 5449)、QSPPT (SEQ ID NO: 5450)、QAKLT (SEQ ID NO: 5451)、KSQQT (SEQ ID NO: 5452)、QDRPT (SEQ ID NO: 5453)、QSQQL (SEQ ID NO: 5454)、QAFHT (SEQ ID NO: 5455)、QKQQD (SEQ ID NO: 5456)、QNAQT (SEQ ID NO: 5457)、HNQLT (SEQ ID NO: 5458)、QNQQY (SEQ ID NO: 5459)、QKLNT (SEQ ID NO: 5460)、QNVQT (SEQ ID NO: 5461)、QAQQT (SEQ ID NO: 5462)、QNLQA (SEQ ID NO: 5463)、QTPPT (SEQ ID NO: 5464)、QYQHA (SEQ ID NO: 5465)、QGQQA (SEQ ID NO: 5466)、QPPAT (SEQ ID NO: 5466) 5467)、QERPT (SEQ ID NO: 5468), QDLQT (SEQ ID NO: 5469), QAMHT (SEQ ID NO: 5470), LNNQQT (SEQ ID NO: 5471), QHPST (SEQ ID NO: 5472), PGLQT (SEQ ID NO: 5473), QGIRT (SEQ ID NO: 5474), QAPAT (SEQ ID NO: 5475), QSQQI (SEQ ID NO: 5476), QIPPT (SEQ ID NO: 5477), QTQLT(SEQ ID NO: 5478)、QAPST (SEQ ID NO: 5479)、QNTYA (SEQ ID NO: 5480)、QNQHI (SEQ ID NO: 5481)、QNHLT (SEQ ID NO: 5482)、QIGMT (SEQ ID NO: 5483)、LNKQT (SEQ ID NO: 5484)、QLQQT (SEQ ID NO: 5485)、QRMST (SEQ ID NO: 5486)、QGILT (SEQ ID NO: 5487)、QDRQT (SEQ ID NO: 5488)、RDWQT (SEQ ID NO: 5489)、QNTHD (SEQ ID NO: 5490)、PNLQI (SEQ ID NO: 5491)、QERST (SEQ ID NO: 5492)、QNYQT (SEQ ID NO: 5493)、QRTCT (SEQ ID NO: 5494)、QIGHT (SEQ ID NO: 5495)、QGAIT (SEQ ID NO: 5496)、QVPPT (SEQ ID NO: 5497)、QVQQI (SEQ ID NO: 5498)、LMRQT (SEQ ID NO: 5499)、QYSVT (SEQ ID NO: 5500)、QAITT (SEQ ID NO: 5501)、QKTLT (SEQ ID NO: 5502)、QNQWT (SEQ ID NO: 5503)、QLHHT (SEQ ID NO: 5504)、QNIII (SEQ ID NO: 5505)、QGHHT (SEQ ID NO: 5506)、QSKVT (SEQ ID NO: 5507)、QLPST (SEQ ID NO: 5508)、IGKQT (SEQ ID NO: 5509)、QAIHT (SEQ ID NO: 5510)、QHGLT (SEQ ID NO: 5511)、QFMCT (SEQ ID NO: 5512)、QHLQT (SEQ ID NO: 5513)、QNHQN (SEQ ID NO: 5514)、QPART (SEQ ID NO: 5515)、QSLQT (SEQ ID NO: 5516)、QSQLT (SEQ ID NO: 5517)、QDRQS (SEQ ID NO: 5518)、QMPST (SEQ ID NO:5519), QGSLT (SEQ ID NO: 5520), QVPAT (SEQ ID NO: 5521), QDKQT (SEQ ID NO: 5522), HYQQT (SEQ ID NO: 5523), QVPST (SEQ ID NO: 5524), RGEQT (SEQ ID NO: 5525), PGQQT (SEQ ID NO: 5526), QSLQI (SEQ ID NO: 5527), LEQQT (SEQ ID NO: 5528), QNQST (SEQ ID NO: 5529), QKVIT (SEQ ID NO: 5530), QNNDQ (SEQ ID NO: 5531), QSVHT (SEQ ID NO: 5532), QPLGT (SEQ ID NO: 5533), HNQET (SEQ ID NO: 5534), QNLQI (SEQ ID NO: 5535), QIQQT (SEQ ID NO: 5536), QVRNT (SEQ ID NO: Description 39 / 390 pages 61 CN 121127597 A 5537), PSNQT (SEQ ID NO: 5538), QVGHT (SEQ ID NO: 5539), QRDIT (SEQ ID NO: 5540), QMPNT (SEQ ID NO: 5541), RGLQT (SEQ ID NO: 5542), QKQQT (SEQ ID NO: 5543), PSLQT (SEQ ID NO: 5544), QRDQT (SEQ ID NO: 5545), QAKGT (SEQ ID NO: 5546), QSAHT (SEQ ID NO: 5547), QSTMT (SEQ ID NO: 5548), QREMT (SEQ ID NO: 5549), QYRAT (SEQ ID NO: 5550), QWQQT (SEQ ID NO: 5551), QRMNT (SEQ ID NO: 5552), GDSQT (SEQ ID NO: 5553), QKIST (SEQ ID NO: 5554), PSMQT (SEQ ID NO: 5555), SPRQT (SEQ ID NO: 5556), MEQQT (SEQ ID NO: 5557), QYQNT (SEQ ID NO: 5558), QHQQT (SEQ ID NO:(ii) any part of the amino acid sequence in (i), such as any 2, 3 or 4 amino acids, such as a sequence of consecutive amino acids; (iii) a sequence of one, two or three but not more than four modified amino acids relative to any of the amino acid sequences in (i); or (iv) an amino acid sequence of one, two or three but not more than four different amino acids relative to any of the amino acid sequences in (i).

[0284] 193. AAV particles as described in embodiments 191 or 192, wherein [N1]-[N2]-[N3]-[N4] are or comprise: (i) an amino acid sequence of any one of SEQ ID NO: 200 or 2887-3076; (ii) an amino acid sequence comprising any part of the amino acid sequence in (i), such as any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 amino acids thereon, such as a sequence of consecutive amino acids; (iii) an amino acid sequence comprising one, two or three but not more than four modified amino acid sequences relative to any one of the amino acid sequences in (i); or (iv) an amino acid sequence comprising one, two or three but not more than four different amino acids relative to any one of the amino acid sequences in (i).

[0285] 194. An AAV particle as described in any one of embodiments 191 to 193, wherein [N1]-[N2]-[N3]-[N4] is or contains GSGSPHSKAQNQQT (SEQ ID NO: 200).

[0286] 195. An AAV particle as described in any one of embodiments 191 to 193, wherein [N1]-[N2]-[N3]-[N4] is or contains VSGSPHSKAQNQQT (SEQ ID NO: 903).

[0287] 196. An AAV particle as described in any one of embodiments 168 to 195, wherein the AAV capsid variant comprises an amino acid other than K (e.g., T, E, or N) at position 449, an amino acid other than T (e.g., S, E, A, N, V, Q, or G) at position 450, an amino acid other than I (e.g., F, E, V, L, D, S, C, T, A, N, H, R, G, or W) at position 451, and / or an amino acid other than N (e.g., I, P, K, R, H, S, M, Q, D, T, L, A, Y, V, F, E, W, or G) at position 452.

[0288] 197. An AAV particle as described in any one of embodiments 168 to 196, wherein the AAV capsid variant comprises amino acid K at position 449, amino acid T at position 450, amino acid I at position 451, and / or amino acid N at position 452, as specified in SEQ ID NO: 138 or 981.

[0289] 198. An AAV particle as described in any one of embodiments 168 to 197, wherein the AAV capsid variant further comprises [NO], wherein [NO] comprises XA, XB, XC, and XD, wherein: (a) XA is K, T, E, or N; (b) XB is T, S, E, A, N, V, Q, or G; (c) XC is I, F, E, V, L, D, S, C, T, A, N, H, R, G, or W; and (d) XD is N, I, P, K, R, H, S, M, Q, D, T, L, A, Y, V, F, E, W, or G; Optionally, the AAV capsid variant comprises an amino acid modification of any of the aforementioned amino acids in (a)-(d), for example, a conservative substitution.

[0290] 199. AAV particles as described in embodiment 198, wherein [NO] is or comprises: (i) KTII (SEQ ID NO: 5565), KTFP (SEQ ID NO: 5566), KTEK (SEQ ID NO: 5567), KTVN (SEQ ID NO: 5568), KTFN (SEQ ID NO: 5569), KTIN (SEQ ID NO: 5570), TTIN (SEQ ID NO: 5571), KSIN (SEQ ID NO: 5572), KTER (SEQ ID NO: 5573), KELH (SEQ ID NO: 5574), KAIN (SEQ ID NO: 5575), KTDN (SEQ ID NO: 5576), KTFH (SEQ ID NO: 5577), KTSN (SEQ ID NO: 5578), ETIN (SEQ ID NO: 5579), NTIN (SEQ ID NO: 5579), KTN (SEQ ID NO: 5570), KTN (SEQ ID NO: 5570), KTN (SEQ ID NO: 5571), KTN (SEQ ID NO: 5572), KTN (SEQ ID NO: 5573), KELH (SEQ ID NO: 5574), KAIN (SEQ ID NO: 5575), KTDN (SEQ ID NO: 5576), KTFH (SEQ ID NO: 5577), KTSN (SEQ ID NO: 5578), ETIN (SEQ ID NO: 5579), NTIN (SEQ ID NO: 5570), KTN ... NO: 5580), KTEN (SEQ ID NO: 5581), KTSS (SEQ ID NO: 5582), KTCN (SEQ ID NO: 5583), KTEH (SEQ ID NO: 5584), KAEM (SEQ ID NO: 5585), KATN (SEQ ID NO: 5586), KAIK (SEQ ID NO: 5587), KTDK (SEQ IDNO: 5588)、KTFK (SEQ ID NO: 5589)、KSDQ (SEQ ID NO: 5590)、KTEI (SEQ ID NO: 5591)、KTID (SEQ ID NO: 5592)、KNTN (SEQ ID NO: 5593)、KTET (SEQ ID NO: 5594)、KTEL (SEQ ID NO: 5595)、KNIN (SEQ ID NO: 5596)、 KTEA (SEQ ID NO: 5597)、KTAN (SEQ ID NO: 5598)、NTIY (SEQ ID NO: 5599)、KTFS (SEQ ID NO: 5600)、KTES (SEQ ID NO: 5601)、KTTN (SEQ ID NO: 5602)、KTED (SEQ ID NO: 5603)、KTNN (SEQ ID NO: 5604)、KEVH (SEQ ID NO: 5605)、KTIS (SEQ ID NO: 5606)、KTVR (SEQ ID NO: 5607)、KTDR (SEQ ID NO: 5608)、ETIK (SEQ ID NO: 5609)、 KNHI (SEQ ID NO: 5610)、KESD (SEQ ID NO: 5611)、KTIK (SEQ ID NO: 5612)、KTDL (SEQ ID NO: 5613)、KTVP (SEQ ID NO: 5614)、KTVI (SEQ ID NO: 5615)、KAEH (SEQ ID NO: 5616)、KNCL (SEQ ID NO: 5617)、KTVK (SEQ ID NO: 5618)、KNAD (SEQ ID NO: 5619)、KTIT (SEQ ID NO: 5620)、KNCV (SEQ ID NO: 5621)、KNAL (SEQ ID NO: 5622)、 KVIN (SEQ ID NO: 5623)、KTEF (SEQ ID NO: 5624)、KTRE (SEQ ID NO: 5625)、KQGE (SEQ ID NO: 5626)、KSEK (SEQ ID NO: 5627)、KNVN (SEQ ID NO: 5628)、KGGE (SEQ ID NO: 5629)、KEFV (SEQ ID NO: 5630)、KSDK (SEQ ID NO:5631), KTEQ (SEQ ID NO: 5632), KEVQ (SEQ ID NO: 5633), KTEY (SEQ ID NO: 5634), KNCW (SEQ ID NO: 5635), KTDV (SEQ ID NO: 5636), KSDI (SEQ ID NO: 5637), KNSI (SEQ ID NO: 5638), KNSL (SEQ ID NO: 5639), KEVV (SEQ ID NO: 5640), KTEP (SEQ ID NO: 5641), KSEL (SEQ ID NO: 5642), KTWQ (SEQ ID NO: 5643), KTEV (SEQ ID NO: 5644), KAVN (SEQ ID NO: 5645), KGVL (SEQ ID NO: 5646), KTEG (SEQ ID NO: 5647), KTRD (SEQ ID NO: 5648), KTGN (SEQ ID NO: 5649), KNAI (SEQ ID NO: 5650), KAEN (SEQ ID NO: 5651), KAET (SEQ ID NO: 5652), KTVH (SEQ ID NO: 5653), KETA (SEQ ID NO: 5654), KNNL (SEQ ID NO: 5655), EAIN (SEQ ID NO: 5656), KSLN (SEQ ID NO: 5657), KTIP (SEQ ID NO: 5658), or KTIH (SEQ ID NO: 5659); (ii) any portion of the amino acid sequence in (i), such as any two or three amino acids therein, such as a sequence of consecutive amino acids; (iii) relative to any one of the amino acid sequences in (i), containing one, two, or three but not more than four modified amino acid sequences; or (iv) Relative to any of the amino acid sequences in (i), an amino acid sequence comprising one, two, or three, but not exceeding four, different amino acids.

[0291] 200. The AAV particle as described in Embodiments 198 or 199, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or comprises: (i) an amino acid sequence of any one of SEQ ID NO: 3239-3526 or 3591-3605; (ii)(i) any portion of the amino acid sequence, such as any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 amino acids thereon, such as a sequence of consecutive amino acids; (iii) a sequence of one, two, or three, but not more than four modified amino acid sequences relative to any of the amino acid sequences in (i); or (iv) a sequence of one, two, or three, but not more than four different amino acids relative to any of the amino acid sequences in (i).

[0292] 201. AAV particles as described in any of embodiments 198 to 200, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or contains KTINGSGSPHSKAQNQQT (SEQ ID NO: 5660).

[0293] 202. AAV particles as described in any one of embodiments 198 to 200, wherein [N0]-[N1]-[N2]-[N3]-[N4] are or contain KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589).

[0294] 203. An adeno-associated virus (AAV) particle comprising an AAV capsid variant (e.g., an AAV9 capsid variant) and a viral genome comprising a β-glucocerebrosidase 1 (GBA1) coding sequence (e.g., encoding human GBA1 protein), wherein the AAV capsid variant comprises an amino acid sequence having the following formula: [N1]-[N2]-[N3] (SEQ ID NO: 6408), wherein: (i) [N1] comprises X1, X2, and X3, wherein X2 is an amino acid other than S and X3 is an amino acid other than G; (ii) [N2] comprises the amino acid sequence SPH; and (iii) [N3] comprises X4, X5, and X6, wherein X4 is K.

[0295] 204. The AAV particle 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, the AAV capsid variant comprises an amino acid modification of any of the aforementioned amino acids in (i) or (ii), for example, a conservative substitution.

[0296] 205. The AAV particle of embodiment 203 or 204, wherein X5 is S and / or X6 is G.

[0297] 206. AAV particles as described in any one of embodiments 203 to 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.

[0298] 207.AAV particles as described in any of embodiments 203 to 206, wherein [N3] is or includes KSG, KIG, KTG, KRG, KSA, KSL, KSE, KSV, KSR, KSW, KSN, KHG, KYG, KSQ, KIV, KSK, KLW, KMG, or KMA.

[0299] 208. AAV particles as described in any of embodiments 203 to 207, wherein [N3] is or includes KSG.

[0300] 209. AAV particles as described in any one of embodiments 203 to 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).

[0301] 210. AAV particles as described in any one of embodiments 203 to 209, wherein [N2]-[N3] are or comprise: Specification 42 / 390 pages 64 CN 121127597 A (i) SPHKSG (SEQ ID NO: 946), SPHKIG (SEQ ID NO: 958), SPHKTG (SEQ ID NO: 4738), SPHKRG (SEQ ID NO: 974), NPHKSG (SEQ ID NO: 5662), SPHKSA (SEQ ID NO: 977), SPHKSL (SEQ ID NO: 4740), SPHKSE (SEQ ID NO: 4741), SPHKSV (SEQ ID NO: 4742), SPHKSR (SEQ ID NO: 951), SPHKSW (SEQ ID NO: 4743), SPHKSN (SEQ ID NO: 4744), SPHKHG (SEQ ID NO: 951), SPHKSW (SEQ ID NO: 4743), SPHKSN (SEQ ID NO: 4744), SPHKHG (SEQ ID NO: 951), SPHKS ...S 4745), SPHKYG (SEQ ID NO: 966), SPHKSQ (SEQ ID NO: 4746), SPHKIV (SEQ ID NO: 5663), SPHKSK (SEQ ID NO: 4747), SPHKLW (SEQ ID NO: 4748), SPHKMG (SEQ ID NO: 4750) or SPHKMA (SEQ IDNO: 4751); (ii) comprising any part of the amino acid sequence in (i), such as any 2, 3, 4 or 5 amino acids thereof, such as an amino acid sequence of consecutive amino acids; (iii) comprising one, two or three but not more than four modified amino acid sequences relative to any of the amino acid sequences in (i); or (iv) comprising one, two or three but not more than four different amino acid sequences relative to any of the amino acid sequences in (i).

[0302] 211. AAV particles as described in any one of embodiments 203 to 210, wherein [N2]-[N3] is or comprises SPHKSG (SEQ ID NO: 946).

[0303] 212. An AAV particle as described in any one of embodiments 203 to 211, wherein the AAV capsid variant comprises an amino acid other than G (e.g., A, K, W, R, L, I, M, N, T, E, Q, Y, H, F, or V) at position 453 according to SEQ ID NO: 138 or 981.

[0304] 213. An AAV particle as described in any one of embodiments 203 to 212, wherein the AAV capsid variant comprises amino acid G at position 453 according to SEQ ID NO: 138 or 981.

[0305] 214. An AAV particle as described in any one of embodiments 203 to 214, wherein: (i) X1 of [N1] is G, A, K, W, R, L, I, M, N, T, E, Q, Y, H, F, or V; (ii) X2 of [N1] is H, Y, R, Q, N, P, or D; (iii) X3 of [N1] is D, E, G, V, or N; Optionally, the AAV capsid variant comprises an amino acid modification of any of the aforementioned amino acids in (i), (ii), or (iii), for example, a conservative substitution.

[0306] 215. An AAV particle as described in any one of embodiments 203 to 214, wherein X2 of [N1] is H and X3 of [N1] is D.

[0307] 216. An AAV particle as described in any one of embodiments 203 to 215, wherein X1 of [N1] is G, X2 of [N1] is H, and X3 of [N1] is D.

[0308] 217. An AAV particle as described in any one of embodiments 203 to 216, wherein [N1] comprises GH, HD, GY, GR, GQ, AH, GN, KH, GP, WH, RH, LH, IH, MH, GD, NH, TH, EH, QH, YH, HH, FH, VH, YD, HE, RG, QD, RD, ND, PD, QV, DD, HN, or NG.

[0309] 218. An AAV capsid variant as described in any one of embodiments 203 to 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.

[0310] 219. An AAV particle as described in any one of embodiments 203 to 218, wherein [N1] is or contains GHD.

[0311] 220. An AAV particle as described in any one of embodiments 203 to 219, wherein [N1]-[N2] contains HDSPH (SEQ ID NO: 4703). Instruction manual 43 / 390 pages 65 CN 121127597 A

[0312] 221. AAV particles as described in any one of embodiments 203 to 220, wherein [N1]-[N2] are or comprise: (i) GHDSPH (SEQ ID NO: 4784), GYDSPH (SEQ ID NO: 4829), GHESPH (SEQ ID NO: 4793), GRGSPH (SEQ ID NO: 4788), GHDNPH (SEQ ID NO: 5664), GQDSPH (SEQ ID NO: 4785), GRDSPH (SEQ ID NO: 4831), AHDSPH (SEQ ID NO: 5665), GNDSPH (SEQ ID NO: 4832), KHDSPH (SEQ ID NO: 5666), GPDSPH (SEQ ID NO: 4833), WHDSPH (SEQ ID NO: 5667), RHDSPH (SEQ ID NO: 5668), LHDSPH (SEQ ID NO: 5669), GQVSPH (SEQ ID NO: 4835), IHDSPH (SEQ ID NO: 5670), MHDSPH (SEQ ID NO: 5671), GDDSPH (SEQ ID NO: 4792), GHNSPH (SEQ ID NO: 4836), NHDSPH (SEQ ID NO: 5672), THDSPH (SEQ ID NO: 5673), GNGSPH (SEQ ID NO: 4805), EHDSPH (SEQ ID NO: 5674), QHDSPH (SEQ ID NO: 5675), YHDSPH (SEQ ID NO: 5676), HHDSPH (SEQ ID NO: 5677), FHDSPH (SEQ ID NO: 5678) or VHDSPH(SEQ ID NO: 5679); (ii) any part of the amino acid sequence in (i), such as any 2, 3, 4 or 5 amino acids therein, such as an amino acid sequence of consecutive amino acids; (iii) an amino acid sequence containing one, two or three but not more than four modified amino acids relative to any of the amino acid sequences in (i); or (iv) an amino acid sequence containing one, two or three but not more than four different amino acids relative to any of the amino acid sequences in (i).

[0313] 222. The AAV particles of any one of embodiments 203 to 221, wherein [N1]-[N2]-[N3] are or include: (i) GHDSPHKSG (SEQ ID NO: 4698), 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), GHDNPHKSG (SEQ ID NO: 5680), GQDSPHKSG (SEQ ID NO: 4908), GHDSPHKSA (SEQ ID NO: 4940), GHDSPHKSL (SEQ ID NO: 5001), GHDSPHKSE (SEQ ID NO: 5003), GRDSPHKSG (SEQ ID NO: 5004), AHDSPHKSG (SEQ ID NO: 5004), AHDSPHKSG (SEQ ID NO: 5005), GHDSPHKSG (SEQ ID NO: 5006), GHDSPHKSG (SEQ ID NO: 5007), GHDSPHKSG (SEQ ID NO: 4998), GHDSPHKSG (SEQ ID NO: 49 ... 5681), GNDSPHKSV (SEQ ID NO: 5005), AHDSPHKIG (SEQ ID NO: 5682), GHESPHKSA (SEQ ID NO: 4939), GQDSPHKIG (SEQ ID NO: 5006), GHDSPHKSV (SEQ ID NO: 5007), GHDSPHKSR (SEQ ID NO: 4942), KHDSPHKSG (SEQ ID NO: 5683), GPDSPHKIG (SEQ ID NO: 5008), GPDSPHKSG (SEQ ID NO: 5009), GHDSPHKSW (SEQ ID NO: 5010), WHDSPHKSG (SEQ ID NO: 5684), RHDSPHKSG (SEQ ID NO: 5685), GHDSPHKSN (SEQ ID NO: 5011), GHDSPHKRG (SEQ ID NO:4937), GHDSPHKHG (SEQ ID NO: 5013), LHDSPHKSG (SEQ ID NO: 5686), GQVSPHKSG (SEQ ID NO: 5014), IHDSPHKSG (SEQ ID NO: 5687), MHDSPHKSG (SEQ ID NO: 5688), GDDSPHKSV (SEQ ID NO: 5015), GHNSPHKSG (SEQ ID NO: 5016), NHDSPHKSG (SEQ ID NO: 5689), THDSPHKSG (SEQ ID NO: 5690), GGNSPHKRG (SEQ ID NO: 5017), EHDSPHKSG (SEQ ID NO: 5691), GHDSPHKYG (SEQ ID NO: 5018), GHDSPKSQ (SEQ ID NO: 5019), QHDSPHKSG (SEQ ID NO: 5692), RHDSPHKIV (SEQ ID NO: (ii) any portion of the amino acid sequence in (i), such as any 2, 3, 4, 5, 6, 7, 8, or 9 amino acids, such as an amino acid sequence of consecutive amino acids; (iii) Relative to any of the amino acid sequences in (i), there is a modified amino acid sequence comprising one, two, or three but no more than four modified amino acids; or (iv) relative to any of the amino acid sequences in (i), there is a modified amino acid sequence comprising one, two, or three but no more than four different amino acids.

[0314] 223. An AAV particle as described in any of embodiments 203 to 222, wherein [N1]-[N2]-[N3] is or comprises GHDSPHKSG (SEQ ID NO: 4698).

[0315] 224. An AAV particle as described in any one of embodiments 203 to 223, wherein the AAV capsid variant comprises an amino acid other than Q (e.g., R, P, H, K, L, V, A, E, or I) at position 456 according to SEQ ID NO: 138, an amino acid other than N (e.g., I, K, S, H, R, T, D, Y, L, W, F, A, Q, or M) at position 457, an amino acid other than Q (e.g., R, V, K, P, Y, H, L, I, E, or M) at position 458, an amino acid other than Q (e.g., H, L, E, P, W, D, I, V, S, K, R, C, M, or N) at position 459, and / or an amino acid other than T (e.g., A, E, K, S, I, P, G, or N) at position 460.

[0316] 225. An AAV particle as described in any one of embodiments 203 to 224, wherein the AAV capsid variant comprises an amino acid other than Q (e.g., R, P, H, K, L, V, A, E, or I) at position 462 according to SEQ ID NO: 982, an amino acid other than N (e.g., I, K, S, H, R, T, D, Y, L, W, F, A, Q, or M) at position 463, an amino acid other than Q (e.g., R, V, K, P, Y, H, L, I, E, or M) at position 464, an amino acid other than Q (e.g., H, L, E, P, W, D, I, V, S, K, R, C, M, or N) at position 465, and / or an amino acid other than T (e.g., A, E, K, S, I, P, G, or N) at position 466.

[0317] 226. An AAV particle as described in any one of embodiments 203 to 225, wherein the AAV capsid variant comprises amino acid Q at position 456, amino acid N at position 457, amino acid Q at position 458, amino acid Q at position 459, and / or amino acid T at position 460, according to SEQ ID NO: 138.

[0318] 227. An AAV particle as described in any one of embodiments 203 to 226, wherein the AAV capsid variant comprises amino acid Q at position 462, amino acid N at position 463, amino acid Q at position 464, amino acid Q at position 465, and / or amino acid T at position 466, according to SEQ ID NO: 138.

[0319] 228. An AAV particle as described in any one of embodiments 203 to 227, wherein the AAV capsid variant further comprises [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, the AAV capsid variant contains an amino acid modification of any of the aforementioned amino acids in (a)-(e), such as a conservative substitution.

[0320] 229. The AAV particle of embodiment 228, wherein [N4] is or comprises: (i) QNQQT (SEQ ID NO: 5412), QIRQT (SEQ ID NO: 5698), QNQHA (SEQ ID NO: 5699), QKQQT (SEQ ID NO: 5543), QSVQT (SEQ ID NO: 5700), RSQQT (SEQ ID NO: 5701), QNKLE (SEQ ID NO: 5702), QNQQK (SEQ ID NO: 5703), QHQQA (SEQ ID NO: 5704), QIQHT (SEQ ID NO: 5705), PRQQT (SEQ ID NO: 5706), HTQQT (SEQ ID NO: 5707), QRQHT (SEQ ID NO: 5708), QSQQT (SEQ ID NO: 5418), QNQQS (SEQ ID NO: 5418), ...IQHT (SEQ ID NO: 5412), Q NO: 5709), RNQET (SEQ ID NO: 5710), QTQLT (SEQ ID NO: 5478), KNQQT (SEQ ID NO: Instructions 45 / 390 Page 67 CN 121127597 A 5711), QDQQT (SEQ ID NO: 5432), HNQQT (SEQ ID NO: 5426), QNQLT (SEQ ID NO: 5423), QTQQT (SEQ ID NO: 5712), QTQQI (SEQ ID NO: 5713), QSKQA (SEQ ID NO: 5714), QNQPP (SEQ ID NO: 5715), QSPQT (SEQ ID NO: 5716), QNYQT (SEQ ID NO: 5493), QNHQT (SEQ ID NO: 5431), QNRQT (SEQ ID NO: 5446), QNQQG (SEQ ID NO: 5717), QNHLT (SEQ ID NO: 5482), QYQHT (SEQID NO: 5447)、QNQWT (SEQ ID NO: 5503)、QNQHT (SEQ ID NO: 5718)、QTRQT (SEQ ID NO: 5719)、QNLHT (SEQ ID NO: 5720)、LNQQT (SEQ ID NO: 5471)、QNQET (SEQ ID NO: 5721)、QHLQT (SEQ ID NO: 5513)、LNQPT (SEQ ID NO: 5722)、QNQDT (SEQ ID NO: 5723)、RNQQT (SEQ ID NO: 5724)、QNLLT (SEQ ID NO: 5725)、QLVIT (SEQ ID NO: 5726)、RTQET (SEQ ID NO: 5727)、QTHQT (SEQ ID NO: 5728)、QNQPA (SEQ ID NO: 5729)、QDQHT (SEQ ID NO: 5730)、QSQHT (SEQ ID NO: 5731)、RNQQI (SEQ ID NO: 5732)、VRQQT (SEQ ID NO: 5733)、QNQHS (SEQ ID NO: 5734)、AWQQT (SEQ ID NO: 5735)、QSVPT (SEQ ID NO: 5736)、QNIQP (SEQ ID NO: 5737)、QNHLN (SEQ ID NO: 5738)、LDQQT (SEQ ID NO: 5739)、PDQQS (SEQ ID NO: 5740)、ESQQT (SEQ ID NO: 5741)、QNKQT (SEQ ID NO: 5742)、QRQLT (SEQ ID NO: 5743)、QIIVT (SEQ ID NO: 5744)、QKQST (SEQ ID NO: 5745)、QSHQT (SEQ ID NO: 5746)、QFVVT (SEQ ID NO: 5747)、QNLQT (SEQ ID NO: 5428)、QNQQI (SEQ ID NO: 5419)、QSQPT (SEQ ID NO: 5748)、QNEQT (SEQ ID NO: 5749)、QSLQT (SEQ ID NO: 5516)、RNRQT (SEQ ID NO: 5750)、QSKQT (SEQ ID NO: 5751)、QNPLT (SEQ ID NO:5752), RDQKT (SEQ ID NO: 5753), HNQQN (SEQ ID NO: 5754), QWKRT (SEQ ID NO: 5755), QSQQI (SEQ ID NO: 5476), QAQQT (SEQ ID NO: 5462), QNHQI (SEQ ID NO: 5756), QNQQA (SEQ ID NO: 5757), QNQLN (SEQ ID NO: 5758), QTQPT (SEQ ID NO: 5759), INQQT (SEQ ID NO: 5560), QKQLT (SEQ ID NO: 5760), RNQLA (SEQ ID NO: 5761), RNQQS (SEQ ID NO: 5762), ISIQT (SEQ ID NO: 5763), QNQQN (SEQ ID NO: 5764), QSQQS (SEQ ID NO: (ii) any part of the amino acid sequence in (i), such as any 2, 3 or 4 amino acids therein, such as a sequence of consecutive amino acids; (iii) a sequence of one, two or three but not more than four modified amino acids relative to any of the amino acid sequences in (i); or (iv) a sequence of one, two or three but not more than four different amino acids relative to any of the amino acid sequences in (i).

[0321] 230. AAV particles as described in embodiments 228 or 229, wherein [N1]-[N2]-[N3]-[N4] are or comprise: (i) an amino acid sequence of any one of SEQ ID NO: 201 or 3160-3237; (ii) any portion of the amino acid sequence in (i), such as any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 amino acids therein, such as a sequence of consecutive amino acids; (iii)Relative to any of the amino acid sequences in (i), comprising one, two, or three but not more than four modified amino acid sequences; or (iv) relative to any of the amino acid sequences in (i), comprising one, two, or three but not more than four different amino acid sequences.

[0322] 231. AAV particles as described in any of embodiments 228 to 230, wherein [N1]-[N2]-[N3]-[N4] is or comprises GHDSPHKSGQNQQT (SEQ ID NO: 201).

[0323] 232. An AAV particle as described in any one of embodiments 203 to 231, wherein the AAV capsid variant comprises an amino acid other than K (e.g., T) at position 449, an amino acid other than T (e.g., A, S, I, V, N, E, Y, C, G, W, or Q) at position 450, an amino acid other than I (e.g., E, V, S, T, N, D, C, G, Q, L, P, A) at position 451, and / or an amino acid other than N (e.g., S, Y, I, K, F, T, D, E, G, V, L, A, M, Q, H, P, or R) at position 452.

[0324] 233. An AAV particle as described in any one of embodiments 203 to 232, wherein the AAV capsid variant comprises amino acid K at position 449, amino acid T at position 450, amino acid I at position 451, and / or amino acid N at position 452, as specified in SEQ ID NO: 138 or 982.

[0325] 234. An AAV particle as described in any one of embodiments 203 to 233, wherein the AAV capsid variant further comprises [NO], wherein [NO] comprises XA, XB, XC and XD, wherein: (a) XA is K or T; (b) XB is T, A, S, I, V, N, E, Y, C, G, W or Q; (c) XC is I, E, V, S, T, N, D, C, G, Q, L, P, A; and (d) XD is N, S, Y, I, K, F, T, D, E, G, V, L, A, M, Q, H, P or R; Optionally, the AAV capsid variant comprises an amino acid modification of any of the aforementioned amino acids in (a)-(d), for example, a conservative substitution.

[0326] 235. AAV particles as described in embodiment 234, wherein [NO] is or comprises: (i) KAIN (SEQ ID NO: 5575), KTIN (SEQ ID NO: 5570), KTES (SEQ ID NO: 5601), TTIN (SEQ ID NO: 5571), KSIN (SEQ ID NO: 5601), KTIN (SEQ ID NO: 5571), KSIN (SEQ ID NO: 5601), KTIN (SEQ ID NO: 5571), KTIN (SEQ ID NO: 5601), KTIN (SEQ ID NO: 5572), KTIN (SEQ ID NO: 5601 ...5572)、KTVN (SEQ ID NO: 5568)、 KSIY (SEQ ID NO: 5775)、KTSN (SEQ ID NO: 5578)、KTTN (SEQ ID NO: 5602)、KIIN (SEQ ID NO: 5776)、KTIS (SEQ ID NO: 5606)、KAII (SEQ ID NO: 5777)、KTIK (SEQ ID NO: 5612)、KTEF (SEQ ID NO: 5624)、KTIT (SEQ ID NO: 5620)、KTNN (SEQ ID NO: 5604)、KTID (SEQ ID NO: 5592)、KAIS (SEQ ID NO: 5778)、KTVD (SEQ ID NO: 5779)、 KTIE (SEQ ID NO: 5780)、KTEG (SEQ ID NO: 5647)、KVIN (SEQ ID NO: 5623)、KAVN (SEQ ID NO: 5645)、KTIY (SEQ ID NO: 5781)、KTDN (SEQ ID NO: 5576)、KTCN (SEQ ID NO: 5583)、KNVV (SEQ ID NO: 5782)、KTEL (SEQ ID NO: 5595)、KTDA (SEQ ID NO: 5783)、KTEV (SEQ ID NO: 5644)、KSEL (SEQ ID NO: 5642)、KTEM (SEQ ID NO: 5784)、 KTEQ (SEQ ID NO: 5632)、KTII (SEQ ID NO: 5565)、KIVN (SEQ ID NO: 5785)、KTEK (SEQ ID NO: 5567)、KTEN (SEQ ID NO: 5581)、KIGN (SEQ ID NO: 5786)、KEVM (SEQ ID NO: 5787)、KYQV (SEQ ID NO: 5788)、KTEA (SEQ ID NO: 5597)、KATN (SEQ ID NO: 5586)、KTEH (SEQ ID NO: 5584)、KTVE (SEQ ID NO: 5789)、KAID (SEQ ID NO: 5790)、 KTIM (SEQ ID NO: 5791)、KEVG (SEQ ID NO: 5792)、KSEM (SEQ ID NO:5793), KAQQ (SEQ ID NO: 5794), KCGE (SEQ ID NO: 5795), KASN (SEQ ID NO: 5796), KTET (SEQ ID NO: 5594), KTIG (SEQ ID NO: 5797), KTDP (SEQ ID NO: 5798), KELV (SEQ ID NO: 47 / 390 page 69 CN 121127597 A 5799), KELM (SEQ ID NO: 5800), KNEI (SEQ ID NO: 5801), KTPN (SEQ ID NO: 5802), KITN (SEQ ID NO: 5803), KTDI (SEQ ID NO: 5804), KTDQ (SEQ ID NO: 5805), KGIN (SEQ ID NO: 5806), KSEI (SEQ ID NO: 5807), KSEK (SEQ ID NO: 5627), KWSA (SEQ ID NO: 5808), KELA (SEQ ID NO: 5809), KQTQ (SEQ ID NO: 5810), KGAD (SEQ ID NO: 5811), KVGE (SEQ ID NO: 5812), KANE (SEQ ID NO: 5813), KTDT (SEQ ID NO: 5814), KTCI (SEQ ID NO: 5815), KELR (SEQ ID NO: 5816), KCQI (SEQ ID NO: 5817), KGVM (SEQ ID NO: 5818), KACD (SEQ ID NO: 5819), KNEL (SEQ ID NO: 5820), KAAE (SEQ ID NO: 5821), KGQN (SEQ ID NO: 5822), KNEF (SEQ ID NO: 5823), KTSI (SEQ ID NO: 5824), KAEH (SEQ ID NO: 5616), KCDQ (SEQ ID NO: 5825), KEIL (SEQ ID NO: 5826), KTER (SEQ ID NO: 5573), KNAI (SEQ ID NO: 5650), KTDK (SEQ ID NO: 5588), KTPD (SEQ ID NO: 5827), KTIH (SEQ ID NO: 5659) or KTEI (SEQ IDNO: 5591); (ii) any part of the amino acid sequence in (i), such as any two or three amino acids thereof, such as an amino acid sequence of consecutive amino acids; (iii) an amino acid sequence containing one, two or three but not more than four modified amino acids relative to any of the amino acid sequences in (i); or (iv) an amino acid sequence containing one, two or three but not more than four different amino acids relative to any of the amino acid sequences in (i).

[0327] 236. The AAV particle of embodiment 234 or 235, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or comprises: (i) an amino acid sequence of any one of SEQ ID NO: 3606-3836; (ii) an amino acid sequence comprising any part of the amino acid sequence of (i), such as any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17 amino acids thereon, such as a sequence of consecutive amino acids; (iii) an amino acid sequence comprising one, two or three but not more than four modified amino acid sequences relative to any one of the amino acid sequences of (i); or (iv) an amino acid sequence comprising one, two or three but not more than four different amino acids relative to any one of the amino acid sequences of (i).

[0328] 237. An AAV particle as described in any one of embodiments 234 to 236, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or contains KTINGHDSPHKSGQNQQT (SEQ ID NO: 5828).

[0329] 238. An AAV particle as described in any one of embodiments 234 to 236, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or contains KAEIGHDSPHKSGQNQQT (SEQ ID NO: 1754).

[0330] 239. An AAV particle as described in any one of embodiments 234 to 236, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or contains KTEKMSGSPHSKAQNQQT (SEQ ID NO: 3241).

[0331] 240. An AAV particle as described in any one of embodiments 168 to 239, wherein [N1]-[N2]-[N3] are present, for example, in ring IV numbered according to SEQ ID NO: 4, 36, 138, 981 or 982.

[0332] 241. An AAV particle as described in any one of embodiments 198 to 202 or 234 to 240, wherein [NO] and [N4] are present, for example, in ring IV numbered according to SEQ ID NO: 4, 36, 138, 981 or 982.

[0333] 242.AAV particles of any one of embodiments 198 to 202 or 234 to 241, wherein [NO] is present immediately after position 448 of the amino acid sequence number according to SEQ ID NO: 4, 36, 138, 981 or 982.

[0334] 243. AAV particles of any one of embodiments 198 to 202 or 234 to 242, wherein [NO] replaces positions 449-452 (e.g., K449, T450, I451 and N452) of SEQ ID NO: 4, 36, 138, 981 or 982 according to page 48 / 390 of the specification 70 CN 121127597 A.

[0335] 244. An AAV particle as described in any one of embodiments 198 to 202 or 234 to 243, wherein [NO] is present immediately after position 448, and wherein [NO] replaces positions 449-452 (e.g., K449, T450, I451, and N452), which are numbered according to SEQ ID NO: 4, 36, 138, 981, or 982.

[0336] 245. An AAV particle as described in any one of embodiments 198 to 202 or 234 to 244, wherein [NO] corresponds to positions 449-452 (e.g., K449, T450, I451, and N452) of any one of SEQ ID NO: 4, 36, 138, 981, or 982.

[0337] 246. An AAV particle as described in any one of embodiments 168 to 245, wherein [N1] is present immediately after position 452, numbered according to SEQ ID NO: 4, 36, 138, 981, or 982.

[0338] 247. An AAV particle as described in any one of embodiments 168 to 246, wherein [N1] replaces positions 453-455 (e.g., G453, S454, and G455) numbered according to SEQ ID NO: 4, 36, 138, 981, or 982.

[0339] 248. An AAV particle as described in any one of embodiments 168 to 246, wherein [N1] replaces position 453 (e.g., G453) numbered according to SEQ ID NO: 4, 36, 138, 981, or 982.

[0340] 249. An AAV particle as described in any one of embodiments 168 to 177, 179 to 181, 183 to 185, 187 to 193, 195 to 200, 202 or 240 to 246, wherein: (i) X1 of [N1] replaces position 453 (e.g., G453); (ii) X2 of [N1] corresponds to position 454 (e.g., S454); and (iii) X3 of [N1] corresponds to position 455 (e.g., G455), wherein (i), (ii) and (iii) are in accordance with SEQ IDNO: 138 or SEQ ID NO: 981.

[0341] 250. An AAV particle as described in any one of embodiments 168 to 176, 178 to 201 or 240 to 246, wherein: (i) X1 of [N1] corresponds to position 453 (e.g., G453); (ii) X2 of [N1] corresponds to position 454 (e.g., S454); and (iii) 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.

[0342] 251. An AAV particle as described in any one of embodiments 203 to 248, wherein: (i) X1 of [N1] corresponds to position 453 (e.g., G453); (ii) X2 of [N1] replaces position 454 (e.g., S454); and (iii) 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.

[0343] 252. An AAV particle as described in any one of embodiments 203 to 248 or 251, wherein [N1] corresponds to positions 453-455 of SEQ ID NO 982 (e.g., G453, H454, D455).

[0344] 253. An AAV particle as described in any one of embodiments 168 to 176, 178 to 201, 240 to 247, or 250, wherein [N1] corresponds to positions 453-455 of SEQ ID NO: 138 or 981 (e.g., G453, S454, G455).

[0345] 254. An AAV particle as described in any one of embodiments 168 to 253, wherein [N2] is present immediately after position 455 numbered according to SEQ ID NO: 4, 36, 138, 981, or 982.

[0346] 255. An AAV particle as described in any one of embodiments 168 to 254, wherein [N2] corresponds to positions 456-458 of SEQ ID NO: 981 or 982 (e.g., S456, P457, and H458).

[0347] 256. An AAV particle as described in any one of embodiments 168 to 254, wherein [N2] corresponds to position 456-458 (e.g., S456, P457, and H458) of any one of SEQ ID NO: 4 or 36-59.

[0348] 257. An AAV particle as described in any one of embodiments 168 to 256, wherein [N2] is present immediately following [N1]. Specification 49 / 390 pages 71 CN 121127597 A

[0349] 258. An AAV particle as described in any one of embodiments 168 to 202, 240 to 247, or 249 to 257, wherein [N3] corresponds to position 459-460 of SEQ ID NO: 981 (e.g., S459, K460, A461).

[0350] 259. An AAV particle as described in any one of embodiments 168 to 202, 240 to 247, or 249 to 257, wherein [N3] corresponds to position 459-460 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 (e.g., S459, K460, A461).

[0351] 260. An AAV particle as described in any one of embodiments 168 to 259, wherein [N2]-[N3] are present immediately after position 455 according to any one of the numbers SEQ ID NO: 4, 36, 138, 981 or 982.

[0352] 261. An AAV particle as described in any one of embodiments 168 to 259, wherein [N2]-[N3] are present immediately after position 455 according to the number SEQ ID NO: 981.

[0353] 262. An AAV particle as described in any one of embodiments 168 to 261, wherein [N2]-[N3] correspond to positions 456-461 of SEQ ID NO: 981 (e.g., S456, P457, H458, S459, K460, A461).

[0354] 263. AAV particles as described in any one of embodiments 168 to 262, wherein [N2]-[N3] correspond to positions 456-461 (e.g., S456, P457, H458, S459, K460, A461) of any one 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.

[0355] 264. AAV particles as described in any one of embodiments 203 to 257 or 259 to 261, wherein [N3] corresponds to positions 459-460 (e.g., K459, S460, G461) of SEQ ID NO: 982.

[0356] 265. An AAV particle as described in any one of embodiments 203 to 257 or 259 to 261, wherein [N3] corresponds to positions 459-460 of SEQ ID NO: 37 (e.g., K459, S460, G461).

[0357] 266. An AAV particle as described in any one of embodiments 203 to 265, wherein [N2]-[N3] immediately follows the position according to SEQ ID NO:The position 455 of number 982 exists.

[0358] 267. AAV particles of any one of embodiments 203 to 246, 248, 252, 255, 257, 260, 263 to 266, wherein [N3] replaces positions 454 and 455 (e.g., S454 and G455) numbered according to SEQ ID NO: 138.

[0359] 268. AAV particles of any one of embodiments 203 to 246, 248, 252, 255, 257, 260, 263 to 267, wherein [N3] exists immediately after [N2] and replaces positions 454 and 455 (e.g., S454 and G455), which are numbered according to SEQ ID NO: 138.

[0360] 269. An AAV particle as described in any one of embodiments 203 to 246, 248, 252, 255, 257, 260, 263 to 268, wherein [N3] is present immediately following [N1]-[N2] and replaces positions 454 and 455 (e.g., S454 and G455), these positions being numbered according to SEQ ID NO: 138.

[0361] 270. An AAV particle as described in any one of embodiments 203 to 257, 260, 264 to 269, wherein [N2]-[N3] corresponds to positions 456-461 of SEQ ID NO: 982 (e.g., S456, P457, H458, K459, S460, G461).

[0362] 271. An AAV particle as described in any one of embodiments 203 to 257, 260, 264 to 270, wherein [N2]-[N3] correspond to positions 456-461 of SEQ ID NO: 37 (e.g., S456, P457, H458, K459, S460, G461).

[0363] 272. An AAV particle as described in any one of embodiments 191 to 202 or 228 to 271, wherein [N4] is present immediately after position 455 according to the numbering in SEQ ID NO: 138.

[0364] 273. AAV particles as described in any one of embodiments 191 to 202 or 228 to 272, wherein [N4] replaces positions 456-460 (e.g., Q456, N457, Q458, Q459, and T460) according to SEQ ID NO: 138.

[0365] 274. AAV particles as described in any one of embodiments 191 to 202 or 228 to 273, wherein [N4] corresponds to positions 462-466 ​​(e.g., Q462, N463, Q464, Q465, and T466) of SEQ ID NO: 981 or 982.

[0366] 275.AAV particles of any one of embodiments 191 to 202 or 228 to 273, wherein [N4] corresponds to position 462-466 ​​of any one of SEQ ID NO: 4 or 36-59 on page 72 of CN 121127597.

[0367] 276. AAV particles of any one of embodiments 191 to 202 or 228 to 274, wherein [N4] corresponds to position 456-460 of SEQ ID NO: 138 (e.g., Q456, N457, Q458, Q459 and T460).

[0368] 277. An AAV particle as described in any one of embodiments 191 to 202 or 228 to 276, wherein [N2]-[N3]-[N4] replaces positions 456-460 (e.g., Q456, N457, Q458, Q459, and T460) numbered according to SEQ ID NO: 138.

[0369] 278. An AAV particle as described in any one of embodiments 191 to 202 or 228 to 277, wherein [N2]-[N3]-[N4] is present immediately after position 455, and wherein [N2]-[N3]-[N4] replaces positions 456-460 (e.g., Q456, N457, Q458, Q459, and T460) numbered according to SEQ ID NO: 138.

[0370] 279. AAV particles as described in any one of embodiments 191 to 202 or 228 to 278, wherein [N1]-[N2]-[N3]-[N4] replace positions 453-460 (e.g., G453, S454, G455, Q456, N457, Q458, Q459 and T460) according to SEQ ID NO: 138.

[0371] 280. AAV particles as described in any of embodiments 191 to 202 or 228 to 279, wherein [N1]-[N2]-[N3]-[N4] are present immediately after position 452, and wherein [N1]-[N2]-[N3]-[N4] replace positions 453-460 (e.g., G453, S454, G455, Q456, N457, Q458, Q459 and T460), which are numbered according to SEQ ID NO: 138.

[0372] 281. AAV particles as described in any one of embodiments 191 to 202, 240 to 247, 249, 250, 253 to 263, 266 or 272 to 280, wherein [N1]-[N2]-[N3]-[N4] corresponds to positions 453-466 of SEQ ID NO: 981 (e.g.,G453, S454, G455, S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465, and T466).

[0373] 282. AAV particles as described in any one of embodiments 168 to 202, 240 to 247, 249, 250, 253 to 263, 266, or 272 to 280, wherein [N1]-[N2]-[N3] correspond to positions 453-461 of SEQ ID NO: 981 (e.g., G453, S454, G455, S456, P457, H458, S459, K460, A461).

[0374] 283. AAV particles as described in any one of embodiments 228 to 257, 260, 261, 264 to 282, wherein [N1]-[N2]-[N3]-[N4] correspond to positions 453-466 of SEQ ID NO: 982 (e.g., G453, H454, D455, S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465, T466).

[0375] 284. AAV particles as described in any one of embodiments 203 to 257, 260, 261, 264 to 283, wherein [N1]-[N2]-[N3] correspond to positions 453-461 of SEQ ID NO: 982 (e.g., G453, H454, D455, S456, P457, H458, K459, S460, G461).

[0376] 285. AAV particles as described in any one of embodiments 228 to 257, 260, 261, 264 to 282, wherein [N1]-[N2]-[N3]-[N4] correspond to positions 453-466 of any one of SEQ ID NO: 4 or 36-59.

[0377] 286. AAV particles as described in any one of embodiments 198 to 202 or 234 to 286, wherein [N0]-[N1]-[N2]-[N3]-[N4] replace positions 449-460 (e.g., K449, T450, I451, N452, G453, S454, G455, Q456, N457, Q458, Q459 and T460) as specified in SEQ ID NO: 138.

[0378] 287. AAV particles as described in any one of embodiments 198 to 202 or 234 to 286, wherein [N0]-[N1]-[N2]-[N3]-[N4] are present immediately after position 448, and wherein [N0]-[N1]-[N2]-[N3]-[N4] replace positions 449-460 according to SEQ ID NO: 138.(For example, K449, T450, I451, N452, G453, S454, G455, Q456, N457, Q458, Q459 and T460).

[0379] 288. AAV particles as described in any one of embodiments 198 to 202, 240 to 247, 249, 250, 253 to 263, 266, 272 to 281, 286 or 287, wherein [N0]-[N1]-[N2]-[N3]-[N4] corresponds to positions 449-466 of SEQ ID NO: 981 (e.g., K449, T450, I451, N452, G453, S454, G455, S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465, T466). Specification page 51 / 390 73 CN 121127597 A

[0380] 289. AAV particles as described in any one of embodiments 234 to 257, 260, 261, 264 to 284, 286 or 287, wherein [N0]-[N1]-[N2]-[N3]-[N4] corresponds to positions 449-466 of SEQ ID NO: 982 (e.g. K449, T450, I451, N452, G453, H454, D455, S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465, T466).

[0381] 290. An AAV particle as described in any one of embodiments 234 to 257, 260, 261, 264 to 284, 286, or 287, wherein [N0]-[N1]-[N2]-[N3]-[N4] correspond to positions 449-466 of any one of SEQ ID NO: 4 or 36-59.

[0382] 291. An AAV particle as described in any one of embodiments 191 to 202 or 228 to 290, wherein [N4] is present immediately after position 461 numbered according to SEQ ID NO: 4, 36, 981, or 982.

[0383] 292. An AAV particle as described in any one of embodiments 191 to 202 or 228 to 291, wherein [N4] replaces positions 462-466 ​​(e.g., Q462, N463, Q464, Q465, and T466) numbered according to SEQ ID NO: 4, 36, 981, or 982.

[0384] 293. An AAV particle as described in any one of embodiments 191 to 202 or 228 to 292, wherein [N2]-[N3]-[N4] replaces positions 462-466 ​​numbered according to SEQ ID NO: 4, 36, 981, or 982.(e.g., Q462, N463, Q464, Q465, and T466).

[0385] 294. An AAV particle as described in any one of embodiments 191 to 202 or 228 to 293, wherein [N2]-[N3]-[N4] are present immediately after position 455, and wherein [N2]-[N3]-[N4] replace positions 462-466 ​​(e.g., Q462, N463, Q464, Q465, and T466), which are numbered according to SEQ ID NO: 4, 36, 981, or 982.

[0386] 295. An AAV particle as described in any one of embodiments 168 to 294, wherein the AAV capsid variant comprises [N2]-[N3] from the N-terminus to the C-terminus.

[0387] 296. An AAV particle as described in any one of embodiments 168 to 295, wherein the AAV capsid variant comprises [N1]-[N2]-[N3] from the N-terminus to the C-terminus.

[0388] 297. An AAV particle as described in any one of embodiments 168 to 296, wherein the AAV capsid variant comprises [N0]-[N1]-[N2]-[N3] from the N-terminus to the C-terminus.

[0389] 298. An AAV particle as described in any one of embodiments 168 to 297, wherein the AAV capsid variant comprises [N1]-[N2]-[N3]-[N4] from the N-terminus to the C-terminus.

[0390] 299. An AAV particle as described in any one of embodiments 168 to 298, wherein the AAV capsid variant comprises [N0]-[N1]-[N2]-[N3]-[N4] from the N-terminus to the C-terminus.

[0391] 300. An adeno-associated virus (AAV) particle comprising an AAV capsid variant (e.g., an AAV9 capsid variant) and a viral genome containing a β-glucocerebrosidase 1 (GBA1) coding sequence (e.g., encoding a human GBA1 protein), wherein the AAV capsid variant comprises 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) X1 is S, I, F, V, C, Y, W, R, P, L, Q, M, K, or G; (b) X2 is K, M, R, F, V, C, P, Y, L, W, G, N, S, T, I, or A; (c) X3 is A, Y, L, R, W, C, T, F, H, I, P, M, K, S, V, G, Q, or N; (d) 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) X5 is A, N, Y, R, K, L, I, M, Q, S, C, W, F, T, G, V, or P;Optionally, the AAV capsid variant comprises an amino acid modification of any of the aforementioned amino acids in (a)-(e), for example, a conservative substitution.

[0392] 301. An AAV particle as described in embodiment 300, wherein: Specification 52 / 390 pages 74 CN 121127597 A (a) X1 is S, L, R, V or P; (b) X2 is K, C, F, L, P, R, S or V; (c) X3 is A, C, F, I, K, L, M, P, R, T, W or Y; (d) X4 is Q, R, S, T, C, F, K, L, P or Y; and (e) X5 is N, R, S, T, K, M, Q or Y; Optionally, the AAV capsid variant comprises an amino acid modification of any of the aforementioned amino acids in (a)-(e), for example, a conservative substitution.

[0393] 302. AAV particles as described in 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.

[0394] 303. An AAV particle as in any one of embodiments 300 to 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), DESCRIPTION 53 / 390 pages 75 CN 121127597 A 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: Description Page 54 / 390 76 CN 121127597 A 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 IDNO: 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: 601)、SEQ ID NO: 601、K NOCT: 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)、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) (SEQ ID NO: 6027)、KAPR ID: 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) 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)、KLTR (SEQ ID NO: 6039)、QLCT 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: 604) (SEQ ID NO: 604) 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)、QPRQ 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).

[0395] 304. AAV particles as described in any one of embodiments 300 to 303, wherein [B] is or comprises: (i) SKAQA (SEQ ID NO: 6102), SKAQN (SEQ ID NO: 6103), SMYMN (SEQ ID NO: 6104), SKAFY (SEQ ID NO: 6105), SKLKR (SEQ ID NO: 6106), SKRHR (SEQ ID NO: 6107), SKAQK (SEQ ID NO: 6108), SKWRL (SEQ ID NO: 6109), SRCRN (SEQ ID NO: 6110), SFTCN (SEQ ID NO: 6111), SKFFI (SEQ ID NO: 6112), SKAQY (SEQ ID NO: 6113), IVWQN (SEQ ID NO: 6114), SKYFM (SEQ ID NO: 6115), SKARQ (SEQ ID NO: 6116), SKAQN (SEQ ID NO: 6113), SKAQN (SEQ ID NO: 6114), SKYFM (SEQ ID NO: 6115), SKARQ (SEQ ID NO: 6116), SKAQN (SEQ ID NO: 6112), SKAQN (SEQ ID NO: 6113), SKAQN (SEQ ID NO: 6114), SKYFM (SEQ ID NO: 6115), SKAQN (SEQ ID NO: 6116), SKAQN (SEQ ID NO: 6114), SKAQN ...5), SKAQN (SEQ ID NO: ID NO: 6116), SCHQN (SEQ ID NO: 6117), FPWQN (SEQ ID NO: 6118), SKIRR (SEQ ID NO: Instruction 55 / 390 Page 77 CN 121127597 A 6119), VYYQN (SEQ ID NO: 6120), SLYWN (SEQ ID NO: 6121), SPKKR (SEQ ID NO: 6122), SKAFS (SEQ ID NO: 6123), SRFWN (SEQ ID NO: 6124), SKAQC (SEQ ID NO: 6125), SRMRN (SEQ ID NO: 6126), SVKKN (SEQ ID NO: 6127), SWAPN (SEQ ID NO: 6128), SLWKN (SEQ ID NO: 6129), SKARW (SEQ ID NO: 6130), SFRPN (SEQ ID NO: 6131), SKKVF (SEQ ID NO: 6132), SYAFN (SEQ ID NO: 6133), SKACS (SEQ ID NO:6134)、SKRLW (SEQ ID NO: 6135)、SCSRN (SEQ ID NO: 6136)、SRCPN (SEQ ID NO: 6137)、SGACN (SEQ ID NO: 6138)、SKFRQ (SEQ ID NO: 6139)、SFPFN (SEQ ID NO: 6140)、SFFGN (SEQ ID NO: 6141)、SMCQN (SEQ ID NO: 6142)、SKLFW (SEQ ID NO: 6143)、SKTRK (SEQ ID NO: 6144)、SGKRN (SEQ ID NO: 6145)、FYRQN (SEQ ID NO: 6146)、CRVQN (SEQ ID NO: 6147)、YGIQN (SEQ ID NO: 6148)、SKAQR (SEQ ID NO: 6149)、VNCQN (SEQ ID NO: 6150)、SKPFR (SEQ ID NO: 6151)、SLAWN (SEQ ID NO: 6152)、SKRSY (SEQ ID NO: 6153)、WSYQN (SEQ ID NO: 6154)、RWLQN (SEQ ID NO: 6155)、PSCQN (SEQ ID NO: 6156)、SSWLN (SEQ ID NO: 6157)、SKRRA (SEQ ID NO: 6158)、SKAQT (SEQ ID NO: 6159)、SKGCT (SEQ ID NO: 6160)、VPWQN (SEQ ID NO: 6161)、SKRYT (SEQ ID NO: 6162)、SGCQN (SEQ ID NO: 6163)、SFTPN (SEQ ID NO: 6164)、STTCN (SEQ ID NO: 6165)、SKARM (SEQ ID NO: 6166)、PKRQN (SEQ ID NO: 6167)、SKCFL (SEQ ID NO: 6168)、WVPQN (SEQ ID NO: 6169)、SFWSN (SEQ ID NO: 6170)、SKFKN (SEQ ID NO: 6171)、RIKQN (SEQ ID NO: 6172)、SKAPR (SEQ ID NO: 6173)、SFRYN (SEQ ID NO: 6174)、SKMIC (SEQ ID NO: 6175)、LRWQN (SEQID NO: 6176)、LPTQN (SEQ ID NO: 6177)、SKWKS (SEQ ID NO: 6178)、SYMRN (SEQ ID NO: 6179)、SKAAR (SEQ ID NO: 6180)、LLCQN (SEQ ID NO: 6181)、RCCQN (SEQ ID NO: 6182)、LCVQN (SEQ ID NO: 6183)、SKLTR (SEQ ID NO: 6184)、SKLCT (SEQ ID NO: 6185)、SKIRG (SEQ ID NO: 6186)、SYLVN (SEQ ID NO: 6187)、QGCQN (SEQ ID NO: 6188)、MAFQN (SEQ ID NO: 6189)、SKACQ (SEQ ID NO: 6190)、SKWGL (SEQ ID NO: 6191)、SKILR (SEQ ID NO: 6192)、SFQIN (SEQ ID NO: 6193)、SKACI (SEQ ID NO: 6194)、SKALR (SEQ ID NO: 6195)、SKAHA (SEQ ID NO: 6196)、SLCLN (SEQ ID NO: 6197)、SKAFV (SEQ ID NO: 6198)、RPWQN (SEQ ID NO: 6199)、RPRQN (SEQ ID NO: 6200)、SCPQN (SEQ ID NO: 6201)、SKAQF (SEQ ID NO: 6202)、SVRYN (SEQ ID NO: 6203)、SVRCN (SEQ ID NO: 6204)、SKMPC (SEQ ID NO: 6205)、SKKTS (SEQ ID NO: 6206)、SLPYN (SEQ ID NO: 6207)、VYHQN (SEQ ID NO: 6208)、SKAQS (SEQ ID NO: 6209)、SYTRN (SEQ ID NO: 6210)、LVYQN (SEQ ID NO: 6211)、YRYQN (SEQ ID NO: 6212)、SWLKN (SEQ ID NO: 6213)、SKAQM (SEQ ID NO: 6214)、CPAQN (SEQ ID NO: 6215)、SMCTN (SEQ ID NO: 6216)、SPPDN (SEQ ID NO:6217), SKRNY (SEQ ID NO: 6218), RWTQN (SEQ ID NO: 6219), SKACR (SEQ ID NO: 6220), PRKQN (SEQ ID NO: 6221), SKCSV (SEQ ID NO: 6222), SKARI (SEQ ID NO: 6223), PFVQN (SEQ ID NO: 6224), SKLPK (SEQ ID NO: 6225), SKSEQ (SEQ ID NO: 6226), WVAQN (SEQ ID NO: 6227), SLYQN (SEQ ID NO: 6228), SKVRM (SEQ ID NO: 6229), CALQN (SEQ ID NO: 6230), SSCTN (SEQ ID NO: 6231), SKNSR (SEQ ID NO: 6232), SKRKR (SEQ ID NO: 6233), LCTQN (SEQ ID NO: 6234), STCLN (SEQ ID NO: 6235), SYARN (SEQ ID NO: Instruction Manual 56 / 390 Page 78 CN 121127597 A 6236), SKQRP (SEQ ID NO: 6237), SKRVV (SEQ ID NO: 6238), KSGQN (SEQ ID NO: 6239), SYYSN (SEQ ID NO: 6240), SLTCN (SEQ ID NO: 6241), SCQSN (SEQ ID NO: 6242), SKAKG (SEQ ID NO: 6243), SKPQN (SEQ ID NO: 6244), FPFQN (SEQ ID NO: 6245), SKCTS (SEQ ID NO: 6246), SVFEN (SEQ ID NO: 6247), SKAKK (SEQ ID NO: 6248) or GRYQN (SEQ ID NO: 6249); (ii) any portion of the amino acid sequence in (i), such as any 2, 3, or 4 amino acids therein, such as an amino acid sequence of consecutive amino acids; (iii) an amino acid sequence containing one, two, or three, but no more than four modified amino acids relative to any of the amino acid sequences in (i); or (iv) an amino acid sequence containing one, two, or three, but no more than four different amino acids relative to any of the amino acid sequences in (i).

[0396] 305.An AAV particle as in any of embodiments 300 to 304, wherein [A]-[B] is or comprises: (i) GSGSPHSKAQA (SEQ ID NO: 6250), GSGSPHSKAQN (SEQ ID NO: 6251), GSGSPHSMYMN (SEQ ID NO: 6252), GSGSPHSKAFY (SEQ ID NO: 6253), GSGSPHSKLKR (SEQ ID NO: 6254), GSGSPHSKRHR (SEQ ID NO: 6255), GSGSPHSKAQK (SEQ ID NO: 6256), GSGSPHSKWRL (SEQ ID NO: 6257), GSGSPHSRCRN (SEQ ID NO: 6258), GSGSPHSFTCN (SEQ ID NO: 6259), GSGSPHSKFFI (SEQ ID NO: 6260), GSGSPHSKAQY (SEQ ID NO: 6261), GSGSPHIVWQN (SEQ ID NO: 6262), GSGSPHSKYFM (SEQ ID NO: 6263), GSGSPHSKARQ (SEQ ID NO: 6264), GSGSPHSCHQN (SEQ ID NO: 6265), GSGSPHFPWQN (SEQ ID NO: 6266), GSGSPHSKIRR (SEQ ID NO: 6267), GSGSPHVYYQN (SEQ ID NO: 6268), GSGSPHSLYWN (SEQ ID NO: 6269), GSGSPHSKPKR (SEQ ID NO: 6270), GSGSPHSKAFS (SEQ ID NO: 6271), GSGSPHSRFWN (SEQ ID NO: 6272), GSGSPHSKAQC (SEQ ID NO: 6273), GSGSPHSRMRN (SEQ ID NO: 6274), GSGSPHSVKKN (SEQ ID NO: 6275), GSGSPHSWAPN (SEQ ID NO: 6276), GSGSPHSLWKN (SEQ ID NO: 6277), GSGSPHSKARW (SEQ ID NO: 6278), GSGSPHSFRPN (SEQ ID NO: 6279), GSGSPHSKKVF (SEQ ID NO: 6280), GSGSPHSYAFN (SEQ IDNO: 6281), GSGSPHSKFRQ (SEQ ID NO: 6287)、GSGSPHSFPFN (SEQ ID NO: 6288)、GSGSPHSFFGN (SEQ ID NO: 6289)、GSGSPHSMCQN (SEQ ID NO: 6290)、GSGSPHSKLFW (SEQ ID NO: 6291)、 GSGSPHSKTRK (SEQ ID NO: 6292)、GSGSPHSGKRN (SEQ ID NO: 6293)、GSGSPHFYRQN (SEQ ID NO: 6294)、GSGSPHCRVQN (SEQ ID NO: 6295)、GSGSPHYGIQN (SEQ ID NO: 6296)、GSGSPHSKAQR (SEQ ID NO: 6297)、GSGSPHVNCQN (SEQ ID NO: 6298)、GSGSPHSKPFR (SEQ ID NO: 6299)、GSGSPHSLAWN (SEQ ID NO: 6300)、GSGSPHSKRSY (SEQ ID NO: 6301)、GSGSPHWSYQN (SEQ ID NO: 6302)、GSGSPHRWLQN (SEQ ID NO: 6303). 6308). 6313)、GSGSPHSKARM (SEQ ID NO:6314), GSGSPHPKRQN (SEQ ID NO: 6315), GSGSPHSKCFL (SEQ ID NO: 6316), Page 57 / 390 of the specification, CN 121127597 A, GSGSPHWVPQN (SEQ ID NO: 6317), GSGSPHSFWSN (SEQ ID NO: 6318), GSGSPHSKFKN (SEQ ID NO: 6319), GSGSPHRIKQN (SEQ ID NO: 6320), GSGSPHSKAPR (SEQ ID NO: 6321), GSGSPHSFRYN (SEQ ID NO: 6322), GSGSPHSKMIC (SEQ ID NO: 6323), GSGSPHLRWQN (SEQ ID NO: 6324), GSGSPHLPTQN (SEQ ID NO: 6325), GSGSPHSKWKS (SEQ ID NO: 6326), GSGSPHSYMRN (SEQ ID NO: 6327), GSGSPHSKAAR (SEQ ID NO: 6328), GSGSPHLLCQN (SEQ ID NO: 6329), GSGSPHRCCQN (SEQ ID NO: 6330), GSGSPHLCVQN (SEQ ID NO: 6331), GSGSPHSKLTR (SEQ ID NO: 6332), GSGSPHSKLCT (SEQ ID NO: 6333), GSGSPHSKIRG (SEQ ID NO: 6334), GSGSPHSYLVN (SEQ ID NO: 6335), GSGSPHQGCQN (SEQ ID NO: 6336), GSGSPHMAFQN (SEQ ID NO: 6337), GSGSPHSKACQ (SEQ ID NO: 6338), GSGSPHSKWGL (SEQ ID NO: 6339), GSGSPHSKILR (SEQ ID NO: 6340), GSGSPHSFQIN (SEQ ID NO: 6341), GSGSPHSKACI (SEQ ID NO: 6342), GSGSPHSKALR (SEQ ID NO: 6343), GSGSPHSKAHA (SEQ ID NO: 6344), GSGSPHSLCLN (SEQ ID NO: 6345), GSGSPHSKAFV (SEQ ID NO:6346)、 GSGSPHRPWQN (SEQ ID NO: 6347)、GSGSPHRPRQN (SEQ ID NO: 6348)、GSGSPHSCPQN (SEQ ID NO: 6349)、GSGSPHSKAQF (SEQ ID NO: 6350)、GSGSPHSVRYN (SEQ ID NO: 6351)、 GSGSPHSVRCN (SEQ ID NO: 6352)、GSGSPHSKMPC (SEQ ID NO: 6353)、GSGSPHSKKTS (SEQ ID NO: 6354)、GSGSPHSLPYN (SEQ ID NO: 6355)、GSGSPHVYHQN (SEQ ID NO: 6356)、 GSGSPHSKAQS (SEQ ID NO: 6357)、GSGSPHSYTRN (SEQ ID NO: 6358)、GSGSPHLVYQN (SEQ ID NO: 6359)、GSGSPHYRYQN (SEQ ID NO: 6360)、GSGSPHSWLKN (SEQ ID NO: 6361)、 GSGSPHSKAQM (SEQ ID NO: 6362)、GSGSPHCPAQN (SEQ ID NO: 6363)、GSGSPHSMCTN (SEQ ID NO: 6364)、GSGSPHSPPDN (SEQ ID NO: 6365)、GSGSPHSKRNY (SEQ ID NO: 6366)、 GSGSPHRWTQN (SEQ ID NO: 6367)、GSGSPHSKACR (SEQ ID NO: 6368)、GSGSPHPRKQN (SEQ ID NO: 6369)、GSGSPHSKCSV (SEQ ID NO: 6370)、GSGSPHSKARI (SEQ ID NO: 6371)、 GSGSPHPFVQN (SEQ ID NO: 6372)、GSGSPHSKLPK (SEQ ID NO: 6373)、GSGSPHSKSEQ (SEQ ID NO: 6374)、GSGSPHWVAQN (SEQ ID NO: 6375)、GSGSPHSLYQN (SEQ ID NO: 6376)、 GSGSPHSKVRM (SEQ ID NO: 6377)、GSGSPHCALQN (SEQ ID NO: 6378)、GSGSPHSSCTN (SEQ ID NO:6379), GSGSPHSKNSR (SEQ ID NO: 6380), GSGSPHSKRKR (SEQ ID NO: 6381), GSGSPHLCTQN (SEQ ID NO: 6382), GSGSPHSTCLN (SEQ ID NO: 6383), GSGSPHSYARN (SEQ ID NO: 6384), GSGSPHSKQRP (SEQ ID NO: 6385), GGSSPHSKRVV (SEQ ID NO: 6386), GGSSPHKSGQN (SEQ ID NO: 6387), GSGSPHSYYSN (SEQ ID NO: 6388), GGSSPHSLTCN (SEQ ID NO: 6389), GSGSPHSCQSN (SEQ ID NO: 6390), GGSSPHSKAKG (SEQ ID NO: 6391), GGSSPHSKPQN (SEQ ID NO: 6392), GGSSPHFPFQN (SEQ ID NO: (ii) any part of the amino acid sequence in (i), such as any 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids, such as a sequence of consecutive amino acids; (iii) a sequence of one, two or three but not more than four modified amino acids relative to any of the amino acid sequences in (i); or (iv) a sequence of one, two or three but not more than four different amino acids relative to any of the amino acid sequences in (i). Specification 58 / 390 pages 80 CN 121127597 A

[0397] 306. An AAV particle as described in any one of embodiments 300 to 305, wherein [A]-[B] does not contain the amino acid sequence of GSGSPHSKAQN (SEQ ID NO: 6251).

[0398] 307. An AAV particle as described in any one of embodiments 300 to 306, wherein the AAV capsid variant contains an amino acid other than Q at position 458 according to SEQ ID NO: 138 (e.g., R, C, S, W, L, F, Y, H, I, V, A orOne, two, or all of the amino acids other than Q at position 459 (e.g., K, I, R, L, or S) and / or the amino acid other than T at position 460 (e.g., R).

[0399] 308. An AAV particle as described in any one of embodiments 300 to 307, wherein the AAV capsid variant comprises: (i) amino acid R at position 458; (ii) amino acid W at position 458; (iii) amino acid Y at position 458; (iv) amino acid F at position 458; (v) amino acid S at position 458; (vi) amino acid C at position 458; (vii) amino acid I at position 458; (viii) amino acid L at position 458; (ix) amino acid P at position 458; (x) amino acid I at position 459; (xi) amino acid H at position 458; or (xii) amino acid V at position 458; wherein (i)-(xii) are numbered according to SEQ ID NO: 138.

[0400] 309. An AAV particle as described in any one of embodiments 300 to 307, wherein the AAV capsid variant comprises: (i) amino acid R at position 458 and amino acid K at position 459; (ii) amino acid C at position 458 and amino acid I at position 459; (iii) amino acid S at position 458 and amino acid R at position 459; (iv) amino acid L at position 458 and amino acid K at position 459; (v) amino acid F at position 458 and amino acid K at position 459; (vi) amino acid C at position 458 and amino acid R at position 459; (vii) amino acid H at position 458 and amino acid R at position 459; (viii) amino acid I at position 458 and amino acid L at position 459; (ix) amino acid V at position 458 and amino acid R at position 459; (x) amino acid A at position 458 and amino acid K at position 459; (xi) Amino acid I at position 458 and amino acid K at position 459; (xii) amino acid C at position 458 and amino acid S at position 459; or (xiii) amino acid C at position 458 and amino acid L at position 459, wherein (i)-(xiii) are numbered according to SEQ ID NO: 138.

[0401] 310. An AAV particle as described in any one of embodiments 300 to 307, wherein the AAV capsid variant comprises amino acid F at position 458, amino acid K at position 459, and amino acid R at position 460, according to SEQ ID NO: 138.

[0402] 311.AAV particles of any one of embodiments 300 to 310, wherein the AAV capsid variant comprises one, two, or all of the amino acids other than T (e.g., Y, P, W, R, K, S, or F) at position 450, the amino acids other than I (e.g., R, S, Y, L, V, H, P, A, or F) at position 451, and / or the amino acids other than N (e.g., V, W, A, T, F, Y, L, R, H, S, or M) at position 452, as per the specification page 59 / 390, 81 CN 121127597 A SEQ ID NO: 138.

[0403] 312. AAV particles of any one of embodiments 300 to 311, wherein the AAV capsid variant comprises amino acid V at position 452, as per the specification page 59 / 390, 81 CN 121127597 A SEQ ID NO: 138.

[0404] 313. An AAV particle as described in any one of embodiments 300 to 312, wherein the AAV capsid variant comprises amino acid Y at position 450 and amino acid V at position 452 according to SEQ ID NO: 138.

[0405] 314. An AAV particle as described in any one of embodiments 300 to 312, wherein the AAV capsid variant comprises amino acid R at position 450 and amino acid Y at position 451 according to SEQ ID NO: 138.

[0406] 315. An AAV particle as described in any one of embodiments 300 to 311, wherein the AAV capsid variant comprises: (i) amino acid P at position 450, amino acid R at position 451, and amino acid W at position 452; (ii) amino acid Y at position 450, amino acid S at position 451, and amino acid A at position 452; (iii) amino acid Y at position 450, amino acid Y at position 451, and amino acid T at position 452; (iv) amino acid P at position 450, amino acid R at position 451, and amino acid F at position 452; (v) amino acid W at position 450, amino acid L at position 451, and amino acid T at position 452; (vi) amino acid R at position 450, amino acid S at position 451, and amino acid Y at position 452; (vii) amino acid Y at position 450, amino acid V at position 451, and amino acid F at position 452; (viii) Amino acid K at position 450, amino acid H at position 451, and amino acid L at position 452; (ix) Amino acid P at position 450, amino acid P at position 451, and amino acid L at position 452; (x) Amino acid P at position 450, amino acid A at position 451, and amino acid R at position 452; (xi) Amino acid S at position 450, amino acid R at position 451, and amino acid R at position 452;(xii) amino acid F at position 450, amino acid F at position 451, and amino acid H at position 452; (xiii) amino acid R at position 450, amino acid F at position 451, and amino acid S at position 452; (xiv) amino acid Y at position 450, amino acid S at position 451, and amino acid M at position 452; or (xv) amino acid P at position 450, amino acid F at position 451, and amino acid L at position 452; wherein (i)-(xv) are numbered according to SEQ ID NO: 138.

[0407] 316. An AAV particle as described in any one of embodiments 300 to 315, wherein the AAV capsid variant comprises: (i) an amino acid sequence of any one of SEQ ID NO: 3849-3982, 2984-4010, 4681-4693; (ii) an amino acid sequence comprising any portion of the amino acid sequence in (i), such as any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17 amino acids therein, such as a sequence of consecutive amino acids; (iii) an amino acid sequence comprising one, two or three but not more than four modified amino acid sequences relative to any one of the amino acid sequences in (i); or (iv) an amino acid sequence comprising one, two or three but not more than four different amino acids relative to any one of the amino acid sequences in (i).

[0408] 317. An AAV particle as described in any one of embodiments 300 to 316, wherein the AAV capsid variant does not contain the amino acid sequence of GSGSPHSKAQNQQ (SEQ ID NO: 1801) or GSGSPHSKAQNQQT (SEQ ID NO: 200).

[0409] 318. An AAV particle as described in any one of embodiments 300 to 317, wherein [A]-[B] are present in ring IV.

[0410] 319. An AAV particle as described in any one of embodiments 300 to 318, wherein [A] is present immediately after position 452 according to SEQ ID NO: 138 or 981. Specification page 60 / 390 82 CN 121127597 A

[0411] 320. An AAV particle as described in any one of embodiments 300 to 319, wherein [A] replaces positions 453-455 (e.g., G453, S454, G455) numbered according to SEQ ID NO: 138 or 981.

[0412] 321. An AAV particle as described in any one of embodiments 300 to 320, wherein [A] is present immediately after position 452, and wherein [A] replaces positions 453-455 (e.g., G453, S454, G455) according to SEQ ID NO: 138.Or number 981.

[0413] 322. An AAV particle as described in any one of embodiments 300 to 321, wherein [B] is present immediately after [A].

[0414] 323. An AAV particle as described in any one of embodiments 300 to 322, wherein [B] replaces positions 456 and 457 (e.g., Q456, N457) according to SEQ ID NO: 138.

[0415] 324. An AAV particle as described in any one of embodiments 300 to 323, wherein [A]-[B] replaces positions 453-457 (e.g., G453, S454, G455, Q456, N457) according to SEQ ID NO: 138.

[0416] 325. An AAV particle as described in any of embodiments 300 to 324, wherein [A]-[B] are present immediately after position 452, and wherein [A]-[B] replace positions 453-457 (e.g., G453, S454, G455, Q456, N457), which are numbered according to SEQ ID NO: 138.

[0417] 326. An AAV particle as described in any of embodiments 300 to 325, wherein the AAV capsid variant comprises [A][B] from the N-terminus to the C-terminus.

[0418] 327. An AAV particle as described in any of the preceding embodiments, wherein the AAV capsid variant comprises at least one, at least two, at least three, or at least four (e.g., 1-4 to 1-5) charged amino acid residues (e.g., acidic and / or basic amino acid residues) relative to SEQ ID NO: 138, present at the N-terminus of the amino acid sequence of SPH (e.g., within 1, 2, 3, 4, 5, or 6 amino acids from the beginning of the amino acid sequence of SPH (e.g., within positions 450-455 according to SEQ ID NO: 138)), optionally, wherein the amino acid sequence of SPH is present at positions 456-458 according to any of SEQ ID NO: 36-59, 981, or 982.

[0419] 328. An AAV particle as described in embodiment 327, wherein the amino acid sequence of SPH is present at positions 456-458 according to any of SEQ ID NO: 36-59, 981, or 982.

[0420] 329. An AAV particle as described in embodiment 327 or 328, wherein the AAV capsid variant comprises fewer than four, fewer than three, or fewer 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.

[0421] 330. An AAV particle as described in any one of embodiments 327 to 329, wherein the AAV capsid variant comprises fewer than four, fewer than three, or fewer 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: 138.138 has a charged amino acid residue (e.g., an acidic or basic amino acid residue) optionally located at any one of positions 450-455 relative to SEQ ID NO: 138.

[0422] 331. An AAV particle as described in any one of embodiments 327 to 330, wherein the charged amino acid residue is an acidic amino acid (e.g., D or E).

[0423] 332. An AAV particle as described in any one of embodiments 327 to 331, wherein the charged amino acid residue is a negatively charged amino acid (e.g., D or E).

[0424] 333. An AAV particle as described in any one of embodiments 327 to 332, wherein the charged amino acid residue is D.

[0425] 334. An AAV particle as described in any one of embodiments 327 to 333, wherein the charged amino acid residue is E.

[0426] 335. An AAV particle as described in any one of embodiments 327 to 334, wherein the charged amino acid residue is a basic amino acid (e.g., K, R, or H).

[0427] 336. An AAV particle as described in any one of embodiments 327 to 335, wherein the charged amino acid residue is a positively charged amino acid (e.g., K, R, or H). Specification 61 / 390 pages 83 CN 121127597 A

[0428] 337. An AAV particle as described in any one of embodiments 327 to 336, wherein the charged amino acid residue is H.

[0429] 338. An AAV particle as described in any one of embodiments 327 to 337, wherein the charged amino acid residue is R.

[0430] 339. An AAV particle as described in any one of embodiments 327 to 338, wherein the charged amino acid residue is K.

[0431] 340. An AAV particle as described in any one of embodiments 327 to 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).

[0432] 341. An AAV particle as described in any one of embodiments 327 to 340, wherein at least one, two, three, or four charged amino acid residues are present in 1, 2, 3, 4, 5, or 6 (e.g., 1-6) amino acids starting from the SPH amino acid sequence.

[0433] 342. An AAV particle as described in any one of embodiments 327 to 341, wherein the AAV capsid variant comprises two charged amino acid residues immediately preceding the SPH amino acid sequence (e.g., at positions 454 and 455 according to SEQ ID NO: 138 or SEQ ID NO: 982).

[0434] 343. An AAV particle as described in any one of embodiments 327 to 342, wherein the AAV capsid variant is contained within a self-

[0435] 344. An AAV particle as described in any one of embodiments 327 to 343, wherein the AAV capsid variant comprises a charged amino acid residue (e.g., E) at position 451 according to any one of SEQ ID NO: 138, 981 or 982.

[0436] 345. An AAV particle as described in any one of embodiments 327 to 344, wherein the AAV capsid variant comprises E at position 451 according to any one of SEQ ID NO: 138, 981 or 982.

[0437] 346. An AAV particle as described in any one of embodiments 327 to 345, wherein the AAV capsid variant comprises a charged amino acid residue (e.g., R or K) at position 452 according to any one of SEQ ID NO: 138, 981 or 982.

[0438] 347. An AAV particle as described in any one of embodiments 327 to 346, wherein the AAV capsid variant comprises R at position 452 according to SEQ ID NO: 138 or SEQ ID NO: 982.

[0439] 348. An AAV particle as described in any one of embodiments 327 to 347, wherein the AAV capsid variant comprises E at position 451 according to SEQ ID NO: 138 or SEQ ID NO: 982 and R at position 452.

[0440] 349. An AAV particle as described in any one of embodiments 327 to 348, wherein the AAV capsid variant has a reduced tropism to hepatocytes or tissues relative to the tropism of an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981.

[0441] 350. An AAV particle as described in any of the preceding embodiments, wherein the AAV capsid variant comprises at least one, at least two, at least three, or at least four (e.g., 1-4 to 1-5) charged amino acid residues (e.g., basic amino acid residues) relative to SEQ ID NO: 138, present at the C-terminus of the amino acid sequence of SPH (e.g., within 1, 2, 3, 4, 5, 6, or 7 amino acids from the end of the amino acid sequence of SPH (e.g., within positions 459-465 according to any one of the numbers in SEQ ID NO: 36-59 or 981)), optionally, wherein the amino acid sequence of SPH is present at positions 456-458 according to any one of the numbers in SEQ ID NO: 36-59, 981, or 982.

[0442] 351. An AAV particle as described in embodiment 350, wherein the amino acid sequence of SPH is present at positions 456-458 according to any one of the numbers in SEQ ID NO:352. The AAV particle of embodiment 350 or 351, wherein the AAV capsid variant comprises fewer than four, fewer than three, fewer 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 particle of embodiment 350 to 352, wherein the AAV capsid variant comprises one charged amino acid residue (e.g., basic amino acid residue) relative to SEQ ID NO: 138, optionally at any of positions 456-460 relative to SEQ ID NO: 138, or at positions 462-466 ​​of any of the numbers in the specification according to SEQ ID NO: 36-59, 981 or 982, page 62 / 390, 84 CN 121127597 A.

[0445] 354. An AAV particle as described in any one of embodiments 350 to 353, wherein the charged amino acid residue is a basic amino acid (e.g., R or K).

[0446] 355. An AAV particle as described in any one of embodiments 350 to 354, wherein the charged amino acid residue is a positively charged amino acid (e.g., R or K).

[0447] 356. An AAV particle as described in any one of embodiments 350 to 355, wherein the charged amino acid residue is R.

[0448] 357. An AAV particle as described in any one of embodiments 350 to 355, wherein the charged amino acid residue is K.

[0449] 358. An AAV particle as described in any one of embodiments 350 to 357, wherein at least one, two, three, or four charged amino acid residues are present in 1, 2, 3, 4, 5, 6, or 7 (e.g., 1-7) amino acids from the end of the SPH amino acid sequence.

[0450] 359. An AAV particle as described in any one of embodiments 350 to 358, wherein the AAV capsid variant comprises a charged amino acid residue (e.g., K or R) immediately following the SPH sequence (e.g., at position 459 according to SEQ ID NO: 981).

[0451] 360. An AAV particle as described in any one of embodiments 350 to 359, wherein the AAV capsid variant comprises a charged amino acid residue (e.g., K or R) at position 459 according to SEQ ID NO: 138 or SEQ ID NO: 982.

[0452] 361. An AAV particle as described in any one of embodiments 350 to 360, wherein the AAV capsid variant comprises K at position 459 according to SEQ ID NO: 981.

[0453] 362.AAV particles of any one of embodiments 350 to 360, wherein the AAV capsid variant contains R at position 459 according to SEQ ID NO: 981.

[0454] 363. AAV particles of any one of embodiments 350 to 362, wherein the AAV capsid variant contains charged amino acid residues (e.g., R or K) at one, two, three, four, five or all of positions 460, 461, 462, 463, 464 and / or 465 according to SEQ ID NO: 138 or 981.

[0455] 364. AAV particles of any one of embodiments 300 to 326 or 350 to 363, wherein the AAV capsid variant has increased tropism to liver cells or tissues relative to the tropism of the AAV capsid containing the amino acid sequence of SEQ ID NO: 138.

[0456] 365. AAV particles as described in any one of embodiments 300 to 326 or 350 to 364, wherein, for example when measured by means of the analysis described in Example 4, the AAV capsid variant is enriched in the liver at at least 10 times, at least 15 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, at least 50 times, at least 55 times, at least 60 times, at least 65 times, at least 70 times, at least 75 times, at least 80 times, at least 85 times, at least 90 times, at least 95 times, at least 100 times, at least 105 times, at least 110 times, at least 115 times, at least 120 times, at least 125 times. At least 130 times, at least 135 times, at least 140 times, at least 150 times, at least 160 times, at least 170 times, at least 180 times, at least 190 times, or at least 200 times.

[0457] 366. An AAV particle as described in any one of embodiments 300 to 326, 364, or 365, wherein the AAV capsid variant has a reduced tropism to CNS cells or tissues, such as brain cells, brain tissue, spinal cord cells, or spinal cord tissue, relative to the tropism of the AAV capsid containing the amino acid sequence SEQ ID NO: 138.

[0458] 367. An AAV particle as described in any one of embodiments 300 to 326 or 364 to 366, wherein the AAV capsid variant exhibits preferential transduction in the liver region relative to transduction in the brain and / or dorsal root ganglion (DRG).

[0459] 368. AAV particles as described in any one of embodiments 300 to 326 or 364 to 367, wherein the AAV capsid variant exhibits preferential transduction in the liver region relative to transduction in the heart and / or muscle (e.g., quadriceps femoris). Specification 63 / 390 pages 85 CN 121127597 A

[0460] 369. An adeno-associated virus (AAV) particle comprising an AAV capsid variant (e.g., the AAV9 capsid variant) and a viral genome containing a β-glucocerebrosidase 1 (GBA1) coding sequence (e.g., encoding the human GBA1 protein), wherein the AAV capsid variant comprises: (a) an amino acid sequence of any one of the sequences provided in Tables 1, 2A, 2B, or 20-26; (b) an amino acid sequence comprising at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, or at least 17 consecutive amino acids from any one of the sequences provided in Tables 1, 2A, 2B, or 20-26; or (c) (d) An amino acid sequence containing at least one, at least two, or at least three but no more than four different amino acids relative to any one of the sequences provided in Tables 1, 2A, 2B, or 20-26; or (d) An amino acid sequence containing at least one, at least two, or at least three but no more than four modified amino acid sequences relative to any one of the sequences provided in Tables 1, 2A, 2B, or 20-26.

[0461] 370. An adeno-associated virus (AAV) particle comprising an AAV capsid variant (e.g., an AAV9 capsid variant) and a viral genome comprising a β-glucocerebrosidase 1 (GBA1) coding sequence (e.g., encoding human GBA1 protein), wherein the AAV capsid variant comprises: (a) an amino acid sequence of any one of SEQ ID NO: 945-980 or 985-986; (b) an amino acid sequence comprising at least 3, at least 4, or at least 5 consecutive amino acids from any one of SEQ ID NO: 945-980 or 985-986; or (c) an amino acid sequence comprising at least one, at least two, or at least three but not more than four different amino acids relative to an amino acid sequence of any one of SEQ ID NO: 945-980 or 985-986; (d) an amino acid sequence comprising at least one, at least two, or at least three but not more than four different amino acids relative to an amino acid sequence of any one of SEQ ID NO: The amino acid sequence of any one of 945-980 or 985-986, comprising at least one, at least two, or at least three but no more than four modified amino acid sequences.

[0462] 371. An adeno-associated virus (AAV) particle comprising an AAV capsid variant (e.g., the AAV9 capsid variant) and a viral genome comprising a β-glucocerebrosidase 1 (GBA1) coding sequence (e.g., encoding human GBA1 protein), wherein the AAV capsid variant comprises: (a) SEQ ID NO:(a) An amino acid sequence comprising any one of SEQ ID NO: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909; (b) An amino acid sequence comprising at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 consecutive amino acids from any one of SEQ ID NO: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909; (c) An amino acid sequence comprising at least one, at least two, or at least three but not more than four different amino acids relative to any one of SEQ ID NO: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909; or (d) An amino acid sequence comprising at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 consecutive amino acids from ... 2. An amino acid sequence of any one of 200, 201, 941, 943, 204, 208, 404 or 903-909, containing at least one, at least two or at least three but no more than four modified amino acid sequences.

[0463] 372. An adeno-associated virus (AAV) particle co...

Claims

1. An adeno-associated virus (AAV) particle comprising: a) an AAV capsid variant comprising an amino acid sequence having the 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; and (iii) [N3] comprises X4, X5, and X6, wherein at least one of X4, X5, or X6 is a basic amino acid; and b) a viral genome comprising a beta-glucocerebrosidase 1 (GBA1) coding sequence.

2. The AAV particle of claim 1, wherein the amino acid sequence [N1]-[N2]-[N3] is in hypervariable loop IV of the AAV capsid variant.

3. The AAV particle of claim 1 or claim 2, wherein the AAV capsid variant is an AAV9 capsid variant.

4. The AAV particle of any one of claims 1 to 3, wherein [N1] comprises X1, X2, and X3, wherein at least one of X1, X2, or X3 is G.

5. The AAV particle of any one of claims 1 to 4, wherein [N2]-[N3] comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941).

6. An adeno-associated virus (AAV) particle comprising: a viral genome comprising a beta-glucocerebrosidase 1 (GBA1) coding sequence; and an AAV9 capsid variant comprising the amino acid sequence of SPHSKA (SEQ ID NO: 941).

7. The AAV particle of claim 6, wherein the amino acid sequence of SPHSKA (SEQ ID NO: 941) is in hypervariable loop IV of the AAV9 capsid variant.

8. The AAV particle of claim 6 or claim 7, wherein the amino acid sequence of SPHSKA (SEQ ID NO: 941) is present immediately after the amino acid position corresponding to position 455 of SEQ ID NO: 4 or SEQ ID NO:

36.

9. The AAV particle of any one of claims 6 to 8, wherein the AAV9 capsid variant further comprises one, two, or all of: N at the amino acid position corresponding to position 452 of SEQ ID NO: 4, E at the amino acid position corresponding to position 451 of SEQ ID NO: 4, and / or V at the amino acid position corresponding to position 453 of SEQ ID NO:

4.

10. The AAV particle of any one of claims 6 to 9, wherein the AAV9 capsid variant comprises the amino acid sequence of K T E N V S G S P H S K A Q N Q Q T (SEQ ID NO: 3272).

11. The AAV particle of any one of claims 6 to 10, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 4; (ii) a VP2 protein comprising an amino acid sequence that is at least 90% identical to positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein comprising an amino acid sequence that is at least 90% identical to positions 203-742 of SEQ ID NO:

4.

12. The AAV particle of any one of claims 6-11, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence that is at least 95% identical to SEQ ID NO: 4; (ii) a VP2 protein comprising an amino acid sequence that is at least 95% identical to positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein comprising an amino acid sequence that is at least 95% identical to positions 203-742 of SEQ ID NO:

4.

13. The AAV particle of any one of claims 6-12, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence that is at least 99% identical to SEQ ID NO: 4; (ii) a VP2 protein comprising an amino acid sequence that is at least 99% identical to positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein comprising an amino acid sequence that is at least 99% identical to positions 203-742 of SEQ ID NO:

4.

14. The AAV particle of any one of claims 6-13, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 4; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO:

4.

15. The AAV particle of any one of claims 6-13, wherein the AAV9 capsid variant comprises: (i) an amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately after the amino acid position corresponding to position 455 of SEQ ID NO: 4; (ii) an E at the amino acid position corresponding to position 451 of SEQ ID NO: 4 and a V at the amino acid position corresponding to position 453 of SEQ ID NO: 4; and (iii) no other modifications relative to wild-type AAV9.

16. The AAV particle of any one of claims 6-8, wherein the AAV9 capsid variant further comprises one, two, or all of: E at the amino acid position corresponding to position 451 of SEQ ID NO: 36, R at the amino acid position corresponding to position 452 of SEQ ID NO: 36, and / or V at the amino acid position corresponding to position 453 of SEQ ID NO:

36.

17. The AAV particle of any one of claims 6-8 and 16, wherein the AAV9 capsid variant comprises the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589).

18. The AAV particle of any one of claims 6-8, 16, and 17, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 36; (ii) a VP2 protein comprising an amino acid sequence that is at least 90% identical to positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein comprising an amino acid sequence that is at least 90% identical to positions 203-742 of SEQ ID NO:

36.

19. The AAV particle of any one of claims 6-8 and 16-18, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence that is at least 95% identical to SEQ ID NO: 36; (ii) a VP2 protein comprising an amino acid sequence that is at least 95% identical to positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein comprising an amino acid sequence that is at least 95% identical to positions 203-742 of SEQ ID NO:

36.

20. The AAV particle of any one of claims 6-8 and 16-19, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence that is at least 99% identical to SEQ ID NO: 36; (ii) a VP2 protein comprising an amino acid sequence that is at least 99% identical to positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein comprising an amino acid sequence that is at least 99% identical to positions 203-742 of SEQ ID NO:

36.

21. The AAV particle of any one of claims 6-8 and 16-20, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 36; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO:

36. (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO:

36.

22. The AAV particle of any one of claims 6-8 and 16-20, wherein the AAV9 capsid variant comprises: (i) the amino acid sequence SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately after the amino acid position corresponding to position 455 of SEQ ID NO: 36; (ii) E at the amino acid position corresponding to position 451 of SEQ ID NO: 36, R at the amino acid position corresponding to position 452 of SEQ ID NO: 36, and V at the amino acid position corresponding to position 453 of SEQ ID NO: 36; and (iii) no other modifications relative to wild-type AAV9.

23. The AAV particle of any one of claims 1-4, wherein [N1]-[N2]-[N3] is present immediately after the position corresponding to amino acid position 452 of SEQ ID NO: 982; and wherein the AAV capsid variant comprises an amino acid sequence that is at least 90% identical, e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 982, e.g., to positions 203-742 of SEQ ID NO:

982.

24. The AAV particle of claim 23, wherein [N1] comprises GHD.

25. The AAV particle of claim 23 or claim 24, wherein [N1] comprises the amino acid G at the position corresponding to position 453 of SEQ ID NO: 138 or SEQ ID NO: 982, the amino acid H at position 454 of SEQ ID NO: 138 or SEQ ID NO: 982, and the amino acid D at position 455.

26. The AAV particle of any one of claims 23-25, wherein [N3] comprises KSG.

27. The AAV particle of any one of claims 23-26, wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence that is at least 90% identical to SEQ ID NO: 982; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 982 or an amino acid sequence that is at least 90% identical to positions 138-742 of SEQ ID NO: 982; or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 982 or an amino acid sequence that is at least 90% identical to positions 203-742 of SEQ ID NO:

982.

28. The AAV particle as claimed in any one of claims 23 to 27, wherein the AAV capsid variant comprises: (i) VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity with SEQ ID NO: 982; (ii) VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity with positions 138-742 of SEQ ID NO: 982; or (iii) VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity with positions 203-742 of SEQ ID NO:

982.

29. The AAV particle as claimed in any one of claims 23 to 28, wherein the AAV capsid variant comprises: (i) VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity with SEQ ID NO: 982; (ii) VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity with positions 138-742 of SEQ ID NO: 982; or (iii) VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity with positions 203-742 of SEQ ID NO:

982.

30. The AAV particle of any one of claims 23 to 29, wherein the AAV capsid variant comprises: (i) VP1 protein, which contains the amino acid sequence of SEQ ID NO: 982; (ii) VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 982; or (iii) VP3 protein, which contains the amino acid sequence at positions 203-742 of SEQ ID NO:

982.

31. The AAV particle according to any one of claims 1 to 30, wherein the viral genome encodes the wild-type GBA1 protein.

32. The AAV particle according to any one of claims 1 to 31, wherein the viral genome does not encode a hemagglutinin (HA) tag.

33. The AAV particle according to any one of claims 1 to 32, wherein the viral genome encodes a human GBA1 protein, a canine GBA1 protein, or a horse GBA1 protein.

34. The AAV particle of claim 33, wherein the viral genome encodes a human GBA1 protein, optionally wherein the human GBA1 protein comprises the amino acid sequence of SEQ ID NO: 1775.

35. The AAV particle according to any one of claims 1 to 34, wherein the GBA1 coding sequence comprises a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO:2002.

36. The AAV particle according to any one of claims 1 to 35, wherein the GBA1 coding sequence comprises at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) of the same nucleotide sequence as SEQ ID NO: 2002.

37. The AAV particle according to any one of claims 1 to 36, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO:2002.

38. The AAV particle according to any one of claims 1 to 37, wherein the GBA1 encoding sequence encodes a signal sequence comprising at least 90% of the same amino acid sequence as SEQ ID NO: 2005.

39. The AAV particle of claim 38, wherein the signal sequence comprises the amino acid sequence of SEQ ID NO: 2005.

40. The AAV particle according to any one of claims 1 to 39, wherein the GBA1 coding sequence comprises a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 2001.

41. The AAV particle of claim 40, wherein the GBA1 encoding sequence comprises SEQ ID NO: 2001.

42. The AAV particle according to any one of claims 1 to 41, wherein the viral genome further comprises a miRNA (miR) binding site that regulates the expression of the encoded GBA1 protein in cells or tissues of the DRG, liver, heart, hematopoietic lineage, or combinations thereof.

43. The AAV particle of any one of claims 1 to 42, wherein the viral genome further comprises a promoter effectively linked to the GBA1 coding sequence.

44. The AAV particle of claim 43, wherein the promoter is at least 90% identical to the nucleotide sequence of SEQ ID NO: 1834 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical).

45. The AAV particle of claim 43, wherein the promoter comprises the nucleotide sequence of SEQ ID NO: 1834.

46. ​​The AAV particle according to any one of claims 1 to 45, wherein the viral genome further comprises an enhancer.

47. The AAV particle of claim 46, wherein the enhancer comprises a CMV immediate early (CMVie) enhancer.

48. The AAV particle of claim 47, wherein the CMVie enhancer is at least 90% identical to the nucleotide sequence of SEQ ID NO: 1831 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical).

49. The AAV particle of claim 47, wherein the CMVie enhancer comprises the nucleotide sequence of SEQ ID NO: 1831.

50. The AAV particle according to any one of claims 1 to 49, wherein the viral genome further comprises introns.

51. The AAV particle of claim 50, wherein the intron is at least 90% identical to the nucleotide sequence of SEQ ID NO: 1842 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical).

52. The AAV particle of claim 50, wherein the intron comprises the nucleotide sequence of SEQ ID NO: 1842.

53. The AAV particle according to any one of claims 1 to 52, wherein the viral genome further comprises a polyadenylated (polyadenylated) region.

54. The AAV particle of claim 53, wherein the polyadenylated region is at least 90% identical to the nucleotide sequence of SEQ ID NO: 1846 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical).

55. The AAV particle of claim 53, wherein the polyadenylate region comprises the nucleotide sequence of SEQ ID NO: 1846.

56. The AAV particle according to any one of claims 1 to 55, wherein the viral genome further comprises inverted terminal repeats (ITRs).

57. The AAV particle of claim 56, wherein the ITR is at least 90% identical to the nucleotide sequence of SEQ ID NO: 1829 or SEQ ID NO: 1830 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical).

58. The AAV particle of claim 56, wherein the ITR comprises the nucleotide sequence of SEQ ID NO: 1829 or SEQ ID NO: 1830.

59. The AAV particle of claim 56, wherein the viral genome comprises a 5' ITR and a 3' ITR, wherein the 5' ITR comprises the nucleotide sequence of SEQ ID NO: 1829 and the 3' ITR comprises the nucleotide sequence of SEQ ID NO: 1830.

60. The AAV particle of any one of claims 1 to 59, wherein the viral genome further comprises one or more miR183 binding sites.

61. The AAV particle of claim 60, wherein the viral genome contains four miR183 binding sites.

62. The AAV particle of claim 61, wherein each of the four miR183 binding sites comprises a nucleotide sequence that is at least 70% identical to the nucleotide sequence of SEQ ID NO: 1847, optionally, wherein each of the four miR183 binding sites comprises the nucleotide sequence of SEQ ID NO: 1847.

63. The AAV particle according to any one of claims 1 to 60, wherein the viral genome further comprises a series of miR183 binding sites, which contain at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) of the nucleotide sequence of SEQ ID NO: 1849.

64. The AAV particle of claim 63, wherein the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 1849.

65. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' reverse terminal repeat (ITR); (ii) Promoter; (iii) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; and (iv) 3' ITR.

66. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' Inverted Terminal Repeat (ITR); (ii) Promoter; (iii) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; and (iv) 3' ITR.

67. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises: (i) 5' Inverted Terminal Repeat (ITR); (ii) CMV immediate early (CMVie) enhancer comprising a nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) Introns containing the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; and (vi) 3' ITR.

68. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises: (i) 5' Inverted Terminal Repeat (ITR); (ii) CMV immediate early (CMVie) enhancer comprising a nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; and (v) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001, and (vi) 3' ITR.

69. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises: (i) 5' Inverted Terminal Repeat (ITR); (ii) A CMV immediate early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1831; (iii) Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) Introns containing the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (vi) A polyadenylated (polyadenylated) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1846; and (vii) 3' ITR.

70. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' Inverted Terminal Repeat (ITR); (ii) CMV immediate early (CMVie) enhancer comprising a nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) Introns containing the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (vi) A polyadenylated (polyadenylated) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1846; and (vii) 3' ITR.

71. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' inverted terminal repeat (ITR) containing the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) CMV immediate early (CMVie) enhancer comprising a nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) Introns containing the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (vi) A polyadenylated (polyadenylated) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1846; and (vii) 3' ITR, which contains the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1830.

72. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises: (i) 5' inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) CMV immediate early (CMVie) enhancer comprising a nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) Introns containing the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (vi) A polyadenylated (polyadenylated) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1846; and (vii) 3' ITR, which contains the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1830.

73. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO:2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 97% identical) to SEQ ID NO:2006.

74. The AAV particle of claim 73, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2006.

75. The AAV particle of claim 74, wherein the viral genome consists of the nucleotide sequence of SEQ ID NO: 2006.

76. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' Inverted Terminal Repeat (ITR); (ii) Promoter; (iii) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (iv) at least one miR183 binding site; and (v) 3' ITR.

77. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' Inverted Terminal Repeat (ITR); (ii) Promoter; (iii) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (iv) at least one miR183 binding site; and (v) 3' ITR.

78. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' Inverted Terminal Repeat (ITR); (ii) Promoter; (iii) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (iv) At least one miR183 binding site sequence comprising at least one miR183 binding site and at least one spacer subsequence; and (v) 3' ITR.

79. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' Inverted Terminal Repeat (ITR); (ii) Promoter; (iii) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (iv) At least one miR183 binding site sequence comprising at least one miR183 binding site and at least one spacer subsequence; and (v) 3' ITR.

80. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' Inverted Terminal Repeat (ITR); (ii) CMV immediate early (CMVie) enhancer comprising a nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) Introns containing the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (vi) At least one miR183 binding site; and (vii) 3' ITR.

81. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' Inverted Terminal Repeat (ITR); (ii) CMV immediate early (CMVie) enhancer comprising a nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) Introns containing the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1842; (v) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (vi) At least one miR183 binding site; and (vii) 3' ITR.

82. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' Inverted Terminal Repeat (ITR); (ii) CMV immediate early (CMVie) enhancer comprising a nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) Introns containing the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (vi) a series of miR183 binding sites comprising the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical to SEQ ID NO: 1849; and (vii) 3' ITR.

83. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' Inverted Terminal Repeat (ITR); (ii) CMV immediate early (CMVie) enhancer comprising a nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831; (iii) Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) Introns containing the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (vi) a series of miR183 binding sites comprising the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1849; and (vii) 3' ITR.

84. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' inverted terminal repeat (ITR) containing the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) CMV immediate early (CMVie) enhancer, which contains the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1831 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iii) Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) Introns containing the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (vi) At least one miR183 binding site comprising the nucleotide sequence of SEQ ID NO: 1847; (vii) A polyadenylated (polyadenylated) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1846; and (viii) 3' ITR, which contains the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1830.

85. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' inverted terminal repeat (ITR) containing the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) CMV immediate early (CMVie) enhancer, which contains the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1831 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iii) Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) Introns containing the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (vi) At least one miR183 binding site comprising the nucleotide sequence of SEQ ID NO: 1847; (vii) A polyadenylated (polyadenylated) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1846; and (viii) 3' ITR, which contains the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1830.

86. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' inverted terminal repeat (ITR) containing the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) CMV immediate early (CMVie) enhancer, which contains the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1831 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iii) Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) Introns containing the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; (vi) miR183 binding site series, which contains the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical to SEQ ID NO: 1849 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (vii) A polyadenylated (polyadenylated) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1846; and (viii) 3' ITR, which contains the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1830.

87. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises, in a 5' to 3' order: (i) 5' inverted terminal repeat (ITR) containing the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829; (ii) CMV immediate early (CMVie) enhancer, which contains the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1831 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iii) Chicken β-actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical to SEQ ID NO: 1834 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (iv) Introns containing the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; (v) A GBA1 coding sequence, wherein the GBA1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence that is at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; (vi) miR183 binding site series, which contains the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical to SEQ ID NO: 1849 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); (vii) A polyadenylated (polyadenylated) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1846; and (viii) 3' ITR, which contains the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1830.

88. The AAV particles as described in claim 87, wherein: (i) The 5' ITR contains the nucleotide sequence of SEQ ID NO: 1829; (ii) The CMVie enhancer contains the nucleotide sequence of SEQ ID NO: 1831; (iii) The CBA promoter contains the nucleotide sequence of SEQ ID NO: 1834; (iv) The intron contains the nucleotide sequence of SEQ ID NO: 1842; (v) The GBA1 coding sequence contains the nucleotide sequence of SEQ ID NO: 2001; (vi) This miR183 binding site series contains the nucleotide sequence of SEQ ID NO: 1849; (vii) The polyadenylated region contains the nucleotide sequence of SEQ ID NO: 1846; and (viii) The 3' ITR contains the nucleotide sequence of SEQ ID NO: 1830.

89. The AAV particle according to any one of claims 1 to 34, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO:2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 97% identical) to SEQ ID NO:2007.

90. The AAV particle of claim 89, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2007.

91. The AAV particle of claim 90, wherein the viral genome consists of the nucleotide sequence of SEQ ID NO: 2007.

92. An adeno-associated virus (AAV) particle comprising a viral genome containing the nucleotide sequence of SEQ ID NO: 2001 and an AAV capsid variant comprising the following: (i) VP1 protein, which contains the amino acid sequence of SEQ ID NO: 4; (ii) VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 4; and / or (iii) VP3 protein, which contains the amino acid sequence at positions 203-742 of SEQ ID NO:

4.

93. An adeno-associated virus (AAV) particle comprising a viral genome containing the nucleotide sequence of SEQ ID NO: 2001 and an AAV capsid variant comprising the following: (i) VP1 protein, which contains the amino acid sequence of SEQ ID NO: 36; (ii) VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 36; and / or (iii) VP3 protein, which contains the amino acid sequence at positions 203-742 of SEQ ID NO:

36.

94. An adeno-associated virus (AAV) particle comprising a viral genome containing the nucleotide sequence of SEQ ID NO: 2002 and an AAV capsid variant comprising the following: (i) VP1 protein, which contains the amino acid sequence of SEQ ID NO: 4; (ii) VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 4; and / or (iii) VP3 protein, which contains the amino acid sequence at positions 203-742 of SEQ ID NO:

4.

95. An adeno-associated virus (AAV) particle comprising a viral genome containing the nucleotide sequence of SEQ ID NO: 2002 and an AAV capsid variant comprising the following: (i) VP1 protein, which contains the amino acid sequence of SEQ ID NO: 36; (ii) VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 36; and / or (iii) VP3 protein, which contains the amino acid sequence at positions 203-742 of SEQ ID NO:

36.

96. A cell comprising AAV particles as claimed in any one of claims 1 to 95, optionally wherein the cell is a mammalian cell (e.g., HEK293 cell), an insect cell (e.g., Sf9 cell), or a bacterial cell.

97. A method for preparing AAV particles as described in any one of claims 1 to 95, the method comprising: (i) Provide a cell containing the viral genome, the viral genome containing the GBA1 coding sequence and nucleic acid encoding the AAV capsid variant; as well as (ii) The cells were cultured under conditions suitable for encapsulating the viral genome within the AAV capsid variant; The AAV particles were prepared in this way.

98. The method of claim 97, wherein the viral genome comprises (a) The nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); or (b) The nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and This AAV capsid variant includes: (i) VP1 protein comprising the amino acid sequence of SEQ ID NO: 4 or an amino acid sequence having at least 90% identity with SEQ ID NO: 4 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity); (ii) VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 4 or having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) with positions 138-742 of SEQ ID NO: 4; or (iii) VP3 protein comprising the amino acid sequence at position 203-742 of SEQ ID NO: 4 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity) with position 203-742 of SEQ ID NO:

4.

99. The method of claim 97, wherein the viral genome comprises (a) The nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); or (b) The nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and The AAV capsid variant contains the amino acid sequence of SEQ ID NO: 4, the amino acid sequence at positions 138-742 of SEQ ID NO: 4, and / or the amino acid sequence at positions 203-742 of SEQ ID NO:

4.

100. The method of claim 97, wherein the viral genome comprises: (a) The nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); or (b) The nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and This AAV capsid variant includes: (i) VP1 protein comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence having at least 90% identity with SEQ ID NO: 36 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity); (ii) VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 36 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) with positions 138-742 of SEQ ID NO: 36; and / or (iii) VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 36 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity) with positions 203-742 of SEQ ID NO:

36.

101. The method of claim 97, wherein the viral genome comprises: (a) The nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); or (b) The nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and The AAV capsid variant contains the amino acid sequence of SEQ ID NO: 36, the amino acid sequence at positions 138-742 of SEQ ID NO: 36, and / or the amino acid sequence at positions 203-742 of SEQ ID NO:

36.

102. The method of claim 97, wherein the viral genome comprises: (a) The nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); or (b) The nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and This AAV capsid variant includes: (i) VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity with SEQ ID NO: 982 (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity); (ii) VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) with positions 138-742 of SEQ ID NO: 982; and / or (iii) VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 982 or having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity) with positions 203-742 of SEQ ID NO:

982.

103. The method of claim 97, wherein the viral genome comprises: (a) The nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); or (b) The nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical); and The AAV capsid variant contains the amino acid sequence of SEQ ID NO: 982, the amino acid sequence at positions 138-742 of SEQ ID NO: 982, and / or the amino acid sequence at positions 203-742 of SEQ ID NO:

982.

104. The method of any one of claims 97 to 103, further comprising introducing a first nucleic acid molecule containing the viral genome into the cell prior to step (i).

105. The method of any one of claims 97 to 104, wherein the cell contains a second nucleic acid molecule encoding the AAV capsid variant.

106. The method of claim 105, further comprising introducing the second nucleic acid into the cell prior to step (i).

107. The method of any one of claims 97 to 106, wherein the cell comprises mammalian cells (e.g., HEK293 cells), insect cells (e.g., Sf9 cells), or bacterial cells.

108. A pharmaceutical composition comprising AAV particles as described in any one of claims 1 to 95 and a pharmaceutically acceptable excipient.

109. A pharmaceutical composition comprising AAV particles as described in any one of claims 5 to 22 and a pharmaceutically acceptable excipient.

110. A pharmaceutical composition comprising AAV particles as described in any one of claims 9 to 15, 92 and 94 and a pharmaceutically acceptable excipient.

111. A pharmaceutical composition comprising AAV particles as described in any one of claims 16 to 22, 93 and 95 and a pharmaceutically acceptable excipient.

112. A method of delivering AAV particles encoding GBA1 protein to a subject, comprising administering to the subject an effective amount of the pharmaceutical composition as described in any one of claims 108 to 111 or the AAV particles as described in any one of claims 1 to 95.

113. The method of claim 112, wherein the subject has, has been diagnosed with, or is at risk of developing a GBA1-related condition, optionally wherein the GBA1-related condition is a GBA1-related neurodegenerative or neuromuscular disease, further optionally wherein the GBA1-related condition is Parkinson's disease (PD), Parkinson's dementia (PDD), Gaucher disease (GD) (e.g., GD type 1, 2, or 3), Lewy body dementia (DLB), Lewy body dementia (LBD), multiple system atrophy (MSA), Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), isolated autonomic failure, brain iron deposition neurodegeneration type 1 (NBIA 1), or Hallewarden-Schpattz syndrome.

114. The method of claim 112 or claim 113, wherein the subject has, has been diagnosed with, or is at risk of developing Parkinson's disease (PD).

115. A method of treating a subject who has or has been diagnosed with GBA1-related conditions, comprising administering to the subject an effective amount of a pharmaceutical composition as described in any one of claims 108 to 111 or AAV particles as described in any one of claims 1 to 95.

116. A method of treating a subject who has or has been diagnosed with GBA1-related conditions, comprising administering to the subject an effective amount of a pharmaceutical composition as described in any one of claims 109 to 111 or AAV particles as described in any one of claims 5 to 22.

117. A method of treating a subject who has or has been diagnosed with GBA1-related conditions, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 110 or AAV particles of any one of claims 9 to 15, 92 and 94.

118. A method of treating a subject who has or has been diagnosed with GBA1-related conditions, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 111 or AAV particles of any one of claims 16 to 22, 93 and 95.

119. The method of any one of claims 115 to 118, wherein the GBA1-related condition is a GBA1-related neurodegenerative or neuromuscular disease, optionally wherein the GBA1-related condition is Parkinson's disease (PD), Parkinson's dementia (PDD), Gaucher disease (GD) (e.g., GD type 1, 2, or 3), dementia with Lewy bodies (DLB), Lewy body dementia (LBD), multiple system atrophy (MSA), Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), isolated autonomic failure, brain iron deposition neurodegeneration type 1 (NBIA 1), or Hallewarden-Scpatz syndrome.

120. The method of claim 119, wherein the GBA1-related neurodegenerative or neuromuscular disorder is Parkinson's disease (PD).

121. The method of claim 119, wherein the GBA1-related neurodegenerative disease or neuromuscular disorder is Gaucher disease (GD) (e.g., GD type 1, 2, or 3).

122. The method of claim 121, wherein the GBA1-related neurodegeneration or neuromuscular disease is GD type 1.

123. The method of claim 121, wherein the GBA1-related neurodegeneration or neuromuscular disease is GD type 2.

124. The method of claim 121, wherein the GBA1-related neurodegeneration or neuromuscular disease is GD type 3.

125. The method of claim 119, wherein the GBA1-related neurodegeneration or neuromuscular disorder is dementia with Lewy bodies (DLB).

126. The method of claim 119, wherein the GBA1-related neurodegeneration or neuromuscular disorder is Lewy body dementia (LBD).

127. A method of treating a subject suffering from Parkinson's disease (PD) or diagnosed with PD, comprising administering to the subject an effective amount of a pharmaceutical composition as described in any one of claims 108 to 111 or AAV particles as described in any one of claims 1 to 95.

128. A method of treating a subject suffering from Parkinson's disease (PD) or diagnosed with PD, comprising administering to the subject an effective amount of a pharmaceutical composition as described in any one of claims 109 to 111 or AAV particles as described in any one of claims 5 to 22.

129. A method of treating a subject with Parkinson's disease (PD) or diagnosed with PD, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 110 or AAV particles of any one of claims 9 to 15, 92 and 94.

130. A method of treating a subject with Parkinson's disease (PD) or diagnosed with PD, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 111 or AAV particles of any one of claims 16 to 22, 93 and 95.

131. A method of treating a subject suffering from or diagnosed with Gaucher disease (GD) (e.g., GD type 1, 2, or 3) or Gaucher disease (GD) (e.g., GD type 1, 2, or 3), comprising administering to the subject an effective amount of the pharmaceutical composition as described in any one of claims 108 to 111 or AAV particles as described in any one of claims 1 to 95.

132. A method of treating a subject suffering from or diagnosed with Gaucher disease (GD) (e.g., GD type 1, 2, or 3) or Gaucher disease (GD) (e.g., GD type 1, 2, or 3), comprising administering to the subject an effective amount of the pharmaceutical composition as claimed in claims 109 to 111 or AAV particles as claimed in any one of claims 5 to 22.

133. A method of treating a subject with Gaucher disease (GD) (e.g., GD type 1, 2, or 3) or diagnosed with Gaucher disease (GD) (e.g., GD type 1, 2, or 3), comprising administering to the subject an effective amount of the pharmaceutical composition of claim 110 or AAV particles as described in any one of claims 9 to 15, 92, and 94.

134. A method of treating a subject with Gaucher disease (GD) (e.g., GD type 1, 2, or 3) or diagnosed with Gaucher disease (GD) (e.g., GD type 1, 2, or 3), comprising administering to the subject an effective amount of the pharmaceutical composition of claim 111 or AAV particles of any one of claims 16 to 22, 93, and 95.

135. The method of any one of claims 131 to 134, wherein the GD is of type GD 1.

136. The method of any one of claims 131 to 134, wherein the GD is of type GD 2.

137. The method of any one of claims 131 to 134, wherein the GD is of type GD 3.

138. A method of treating a subject with Lewy body dementia (LBD), comprising administering to the subject an effective amount of a pharmaceutical composition as described in any one of claims 108 to 111 or AAV particles as described in any one of claims 1 to 95.

139. A method of treating a subject with Lewy body dementia (LBD), comprising administering to the subject an effective amount of the pharmaceutical composition as claimed in claims 109 to 111 or AAV particles as claimed in any one of claims 5 to 22.

140. A method of treating a subject with Lewy body dementia (LBD) comprising administering to the subject an effective amount of the pharmaceutical composition as claimed in claim 111 or AAV particles as claimed in any one of claims 9 to 15, 92 and 94.

141. A method of treating a subject with Lewy body dementia (LBD) comprising administering to the subject an effective amount of the pharmaceutical composition as claimed in claim 111 or AAV particles as claimed in any one of claims 16 to 22, 93 and 95.

142. A method of treating a subject with dementia having Lewy bodies (DLB), comprising administering to the subject an effective amount of a pharmaceutical composition as described in any one of claims 108 to 111 or AAV particles as described in any one of claims 1 to 95.

143. A method of treating a subject with dementia having Lewy bodies (DLB), comprising administering to the subject an effective amount of the pharmaceutical composition as claimed in claims 109 to 111 or AAV particles as claimed in any one of claims 5 to 22.

144. A method of treating a subject with dementia having Lewy bodies (DLB), comprising administering to the subject an effective amount of the pharmaceutical composition as claimed in claim 111 or AAV particles as claimed in any one of claims 9 to 15, 92 and 94.

145. A method of treating a subject with dementia having Lewy bodies (DLB), comprising administering to the subject an effective amount of the pharmaceutical composition as claimed in claim 111 or AAV particles as claimed in any one of claims 16 to 22, 93 and 95.

146. The method of any one of claims 112 to 145, wherein the subject has one or more mutations in the GBA1 gene.

147. The method of any one of claims 112 to 146, wherein the subject has lower GC enzyme activity compared to the subject without GBA1-related disease, optionally wherein the level of GC enzyme activity is measured by a 4-MUG assay or a SensoLyte blue glucocerebrosidase assay.

148. The method of any one of claims 115 to 147, wherein the treatment prevents the progression of the disease in the subject.

149. The method of any one of claims 115 to 148, wherein the treatment improves at least one symptom of the condition and / or alters one or more biomarkers of the condition.

150. The method of claim 149, wherein the one or more biomarkers comprise GC enzyme activity, levels of glucocerebroside and other glycolipids (e.g., in immune cells, such as macrophages), levels of synuclein aggregates (e.g., Lewy bodies), levels of neurofilament light chains, or combinations thereof.

151. The method of claim 149, wherein the at least one symptom comprises developmental delay, progressive encephalopathy, progressive dementia, ataxia, myoclonus, oculomotor dysfunction, bulbar palsy, general weakness, limb tremor, depression, visual hallucinations, cognitive decline, or a combination thereof.

152. The method of any one of claims 112 to 151, wherein the subject is a human.

153. The method of any one of claims 112 to 152, wherein the AAV particles are delivered to cells, tissues, or regions of the CNS, such as regions of the brain or spinal cord, such as the parenchyma, cortex, substantia nigra, caudate nucleus, cerebellum, striatum, corpus callosum, cerebellum, brainstem caudate nucleus-putamen, thalamus, superior colliculus, spinal cord, or combinations thereof, wherein the AAV particles or pharmaceutical composition are delivered via intravenous administration.

154. The method of any one of claims 112 to 153, further comprising assessing, for example, measuring the level of GBA1 expression in the subject, such as the level of GBA1 gene expression, the level of GBA1 mRNA expression, and / or the level of GBA1 protein expression in the subject's cells, tissues, or body fluids.

155. The method of any one of claims 112 to 154, wherein the expression level of the GBA1 protein is measured by ELISA, Western blotting, or immunohistochemistry.

156. The method of claim 154 or claim 155, wherein the assessment of the GBA1 expression level is performed before and after administration of the AAV, optionally wherein the GBA1 expression level before administration is compared with the GBA1 expression level after administration.

157. The method of any one of claims 154 to 156, comprising assessing the expression level of GBA1 in cells or tissues of the central nervous system (e.g., parenchyma).

158. The method of claim 156 or claim 157, wherein the subject's GBA1 protein expression level increases after administration relative to the subject's GBA1 protein expression level prior to administration.

159. The method of any one of claims 112 to 158, further comprising, for example, evaluating, a measure of the GC enzyme activity level in the subject.

160. The method of any one of claims 112 to 159, wherein the application results in the following increase: (i) GC enzyme activity in the subject’s cells, tissues (e.g., cells or tissues of the CNS, such as the cortex, striatum, thalamus, cerebellum, and / or brainstem) and / or body fluids (e.g., CSF and / or serum), optionally, wherein the GC enzyme activity in the subject increases by at least 2, at least 3, at least 4, or at least 5 times after the administration relative to the GC enzyme activity in the subject before the administration; (ii) The level of viral genome (VG) in each cell of the subject's CNS tissue (e.g., the cortex, striatum, thalamus, cerebellum, brainstem, and / or spinal cord), optionally wherein the level of VG in each cell of the subject's CNS tissue is increased by more than 50 VG per cell relative to the level of VG in each cell of the subject's peripheral tissue; and / or (iii) GBA1 mRNA expression in the subject’s cells or tissues (e.g., cells or tissues of the CNS, such as the cortex, thalamus, and / or brainstem), optionally wherein GBA1 mRNA expression in the subject increases by at least 100-1300 times after administration compared to GBA1 mRNA expression in the subject before administration.

161. The method of any one of claims 115 to 160, further comprising administering to the subject an additional agent suitable for treating or preventing the GBA1-related condition; Optionally, the additional agent comprises enzyme replacement therapy (ERT) (e.g., imiglucerase, veralucerase α, or taliglucerase α); substrate reduction therapy (SRT) (e.g., elixstat or miglustat), levodopa, carbidopa, safenamide, dopamine agonists (e.g., quetiapine, clozapine, pramipexole, rotigotine, or ropinirole), anticholinergics (e.g., benzalkonium chloride or trihexyphenidyl), cholinesterase inhibitors (e.g., rivastigmine, donepezil, or galantamine), N-methyl-d-aspartate (NMDA) receptor antagonists (e.g., memantine), or combinations thereof.

162. The method of any one of claims 112 to 161, further comprising administering a blood transfusion to the subject.

163. The method of any one of claims 112 to 161, further comprising administering an immunosuppressant to the subject.

164. The method of claim 162, wherein the immunosuppressant comprises a corticosteroid (e.g., prednisone, prednisolone, methylprednisolone, and / or dexamethasone), rapamycin, mycophenolate mofetil, tacrolimus, rituximab, and / or ikuzumab hydroxychloroquine.

165. The pharmaceutical composition of any one of claims 108 to 111 or the AAV particles of any one of claims 1 to 95, for the treatment of GBA1-related conditions; optionally, wherein the GBA1-related condition is Parkinson's disease (PD), Parkinson's dementia (PDD), Gaucher disease (GD) (e.g., GD type 1, 2, or 3), Lewy body dementia (LBD), Lewy body dementia (LBD), multiple system atrophy (MSA), Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), isolated autonomic failure, brain iron deposition neurodegeneration type 1 (NBIA 1), or Hallewarden-Scpatz syndrome.

166. The pharmaceutical composition or AAV particles of claim 165, wherein the GBA1-related condition is Gaucher disease (GD) (e.g., GD type 1, 2, or 3).

167. The pharmaceutical composition or AAV particles of claim 166, wherein the GD is GD type 1.

168. The pharmaceutical composition or AAV particles of claim 166, wherein the GD is GD type 2.

169. The pharmaceutical composition or AAV particles of claim 166, wherein the GD is GD type 3.

170. The pharmaceutical composition or AAV particles of claim 165, wherein the GBA1-related condition is Parkinson's disease (PD).

171. The pharmaceutical composition or AAV particles of claim 165, wherein the GBA1-related condition is LBD.

172. The pharmaceutical composition or AAV particles of claim 165, wherein the GBA1-related condition is DLB.

173. Use of a pharmaceutical composition as described in any one of claims 108 to 111 or AAV particles as described in any one of claims 1 to 95 in the preparation of a medicament for treating GBA1-related conditions; optionally, wherein the GBA1-related condition is Parkinson's disease (PD), Parkinson's dementia (PDD), Gaucher disease (GD) (e.g., GD type 1, 2, or 3), dementia with Lewy bodies (DLB), Lewy body dementia (LBD), multiple system atrophy (MSA), Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), isolated autonomic failure, brain iron deposition neurodegeneration type 1 (NBIA 1), or Hallewarden-Scpatz syndrome.

174. The use as described in claim 173, wherein the GBA1-related condition is PD.

175. The use as described in claim 173, wherein the GBA1-related condition is Gaucher disease (GD) (e.g., GD type 1, 2, or 3).

176. The use as described in claim 175, wherein the GD is type GD 1.

177. The use as described in claim 175, wherein the GD is type GD 2.

178. The use as described in claim 175, wherein the GD is type GD 3.

179. The use as described in claim 173, wherein the GBA1-related condition is LBD.

180. The use as described in claim 173, wherein the GBA1-related condition is DLB.