Capsid variants and methods of using the same
The disclosure is directed in part to variant capsid polypeptides that can be used to deliver payloads.
1. A variant adeno-associated virus (AAV) capsid polypeptide having at least 95% sequence identity to SEQ ID NO: 2, wherein the variant AAV capsid polypeptide comprises:
(a) an alanine at a position corresponding to W595 as compared to SEQ ID NO: 1
(b) a leucine at a position corresponding to V596 as compared to SEQ ID NO: 1; and
(c) a serine at a position corresponding to N598 as compared to SEQ ID NO: 1.
2. The variant AAV capsid polypeptide of claim 1 , further comprising a valine at a position corresponding to Q579 as compared to SEQ ID NO: 1 and optionally a valine at a position corresponding to T593 as compared to SEQ ID NO: 1.
3. The variant AAV capsid polypeptide of claim 1 , further comprising:
a valine at a position corresponding to Q579 as compared to SEQ ID NO: 1;
an isoleucine at a position corresponding to Q592 as compared to SEQ ID NO: 1;
a valine at a position corresponding to T593 as compared to SEQ ID NO: 1; and
an alanine at a position corresponding to 1601 as compared to SEQ ID NO: 1.
4. The variant AAV capsid polypeptide of claim 1 , comprising the mutation set of any one of SEQ ID NOs: 2, 14, 16, 18, 19, 21, 23, 24, 31, 34, 37, 38, 62, 98 and 128.
5. The variant AAV capsid polypeptide of claim 1 , comprising the amino acid sequence of any one of SEQ ID NOs: 2, 14, 16, 18, 19, 21, 23, 24, 31, 34, 37, 38, 62, 72, 98 and 128.
6. The variant AAV capsid polypeptide of claim 3 , which has at least 96% sequence identity to SEQ ID NO: 2.
7. The variant AAV capsid polypeptide of claim 3 , which has at least 98% sequence identity to SEQ ID NO: 2.
8. The variant AAV capsid polypeptide of claim 3 , which has at least 99% sequence identity to SEQ ID NO: 2.
9. A variant AAV capsid polypeptide comprising the amino acid sequence of SEQ ID NO: 2.
10. A recombinant AAV virus particle comprising the variant AAV capsid polypeptide of claim 1 .
11. A recombinant AAV virus particle comprising the variant AAV capsid polypeptide of claim 3 .
12. A recombinant AAV virus particle comprising the variant AAV capsid polypeptide of claim 9 .
13. The recombinant AAV virus particle of claim 10 , comprising a nucleic acid molecule comprising a promoter operably linked to a heterologous transgene.
14. The recombinant AAV virus particle of claim 11 , comprising a nucleic acid molecule comprising a promoter operably linked to a heterologous transgene.
15. The recombinant AAV virus particle of claim 12 , comprising a nucleic acid molecule comprising a promoter operably linked to a heterologous transgene.
16. An isolated host cell comprising the variant AAV capsid polypeptide of claim 1 .
17. An isolated host cell comprising the variant AAV capsid polypeptide of claim 3 .
18. An isolated host cell comprising the variant AAV capsid polypeptide of claim 9 .
19. An isolated host cell comprising the recombinant AAV virus particle of claim 12 .
20. The variant AAV capsid polypeptide of claim 2 , which comprises a valine at a position corresponding to T593 as compared to SEQ ID NO: 1.
21. The variant AAV capsid polypeptide of claim 1 , which has at least 96% sequence identity to SEQ ID NO: 2.
22. The variant AAV capsid polypeptide of claim 1 , which has at least 98% sequence identity to SEQ ID NO: 2.
23. The variant AAV capsid polypeptide of claim 1 , which has at least 99% sequence identity to SEQ ID NO: 2.
24. The variant AAV capsid polypeptide of claim 4 , which has at least 98% sequence identity to SEQ ID NO: 2.
25. The variant AAV capsid polypeptide of claim 4 , which has at least 99% sequence identity to SEQ ID NO: 2.
26. A recombinant AAV virus particle comprising the variant AAV capsid polypeptide of claim 22 .
27. A recombinant AAV virus particle comprising the variant AAV capsid polypeptide of claim 24 .
28. The recombinant AAV virus particle of claim 26 , comprising a nucleic acid molecule comprising a promoter operably linked to a heterologous transgene.
29. The recombinant AAV virus particle of claim 27 , comprising a nucleic acid molecule comprising a promoter operably linked to a heterologous transgene.
30. A recombinant AAV comprising:
(a) an AAV capsid comprising AAV capsid polypeptides, each having an amino acid sequence consisting of the amino acid sequence of SEQ ID NO: 2, and
(b) a minigene having AAV inverted terminal repeats and a transgene comprising a heterologous gene operably linked to regulatory sequences which direct expression of the heterologous gene in a host cell.