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 .- 49 . (canceled)
50 . A method of delivering a payload to a cell comprising contacting the cell with a recombinant adeno-associated virus (AAV) virus particle comprising (1) a nucleic acid molecule comprising a payload and one or more regulatory elements, and (2) a variant AAV capsid polypeptide having at least 95% sequence identity to SEQ ID NO: 2 and comprising:
(a) a valine at a position corresponding to Q579 as compared to SEQ ID NO: 1 or a leucine at a position corresponding to V596 as compared to SEQ ID NO: 1; and
(b) at least two of the group consisting of:
(i) an isoleucine at a position corresponding to Q592 as compared to SEQ ID NO: 1;
(ii) a valine at a position corresponding to T593 as compared to SEQ ID NO: 1;
(iii) an alanine at a position corresponding to W595 as compared to SEQ ID NO: 1;
(iv) a serine at a position corresponding to N598 as compared to SEQ ID NO: 1; and
(v) an alanine at a position corresponding to I601 as compared to SEQ ID NO: 1.
51 . The method of claim 50 , wherein the variant AAV capsid polypeptide comprises:
(a) a leucine at a position corresponding to V596 as compared to SEQ ID NO: 1, a serine at a position corresponding to N598 as compared to SEQ ID NO: 1, and an alanine at a position corresponding to I601 as compared to SEQ ID NO: 1;
(b) a valine at a position corresponding to Q579 as compared to SEQ ID NO: 1, a leucine at a position corresponding to V596 as compared to SEQ ID NO: 1, a serine at a position corresponding to N598 as compared to SEQ ID NO: 1, and an alanine at a position corresponding to I601 as compared to SEQ ID NO: 1;
(c) an alanine at a position corresponding to W595 as compared to SEQ ID NO: 1, a leucine at a position corresponding to V596 as compared to SEQ ID NO: 1, and a serine at a position corresponding to N598 as compared to SEQ ID NO: 1;
(d) a valine at a position corresponding to T593 as compared to SEQ ID NO:
1, an alanine at a position corresponding to W595 as compared to SEQ ID NO: 1, a leucine at a position corresponding to V596 as compared to SEQ ID NO: 1, and a serine at a position corresponding to N598 as compared to SEQ ID NO: 1;
(e) 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 a leucine at a position corresponding to V596 as compared to SEQ ID NO: 1;
(f) a valine at a position corresponding to T593 as compared to SEQ ID NO: 1, a leucine at a position corresponding to V596 as compared to SEQ ID NO: 1, a serine at a position corresponding to N598 as compared to SEQ ID NO: 1, and an alanine at a position corresponding to I601 as compared to SEQ ID NO: 1;
(g) a valine at a position corresponding to Q579 as compared to SEQ ID NO: 1, an alanine at a position corresponding to W595 as compared to SEQ ID NO: 1, a leucine at a position corresponding to V596 as compared to SEQ ID NO: 1, and a serine at a position corresponding to N598 as compared to SEQ ID NO: 1;
(h) a valine at a position corresponding to Q579 as compared to SEQ ID NO: 1; or
(i) a valine at a position corresponding to Q579 as compared to SEQ ID NO: 1, a valine at a position corresponding to T593 as compared to SEQ ID NO: 1, an alanine at a position corresponding to W595 as compared to SEQ ID NO: 1, a leucine at a position corresponding to V596 as compared to SEQ ID NO: 1, and a serine at a position corresponding to N598 as compared to SEQ ID NO: 1.
52 . The method of claim 50 , wherein the variant AAV capsid polypeptide comprises the mutation set of any one of SEQ ID NOs: 2, 14-39, 42, 44-45, 49, 51, 53, 55-57, 59, 61-63, 66-68, 70-72, 74-75, 78, 82-91, 93, 96, 98, 100, 102-105, 118-121, 123-124, 128-136, or 138.
53 . The method of claim 50 , wherein the variant AAV capsid polypeptide comprises any one of SEQ ID NOs: 2, 14-39, 42, 44-45, 49, 51, 53, 55-57, 59, 61-63, 66-68, 70-72, 74-75, 78, 82-91, 93, 96, 98, 100, 102-105, 118-121, 123-124, 128-136, or 138.
54 . The method of claim 50 , wherein the variant AAV capsid polypeptide has at least 96% sequence identity to SEQ ID NO: 2.
55 . The method of claim 50 , wherein the variant AAV capsid polypeptide has at least 98% sequence identity to SEQ ID NO: 2.
56 . The method of claim 50 , wherein the variant AAV capsid polypeptide has at least 99% sequence identity to SEQ ID NO: 2.
57 . The method of claim 50 , wherein the cell is a central nervous system (CNS) cell.
58 . The method of claim 57 , wherein the CNS cell is a neuronal cell, a glial cell, an astrocyte, an oligodendrocyte, or an endothelial cell.