Recombinant adeno-associated virus vectors
AAV capsid proteins comprising a modification in the amino acid sequence and virus vectors comprising the modified AAV capsid protein are described. Also described are methods of administering the virus vectors and virus capsids to a cell or to a subject in vivo.
1. An adeno-associated virus (AAV) vector comprising an AAV capsid protein, wherein the AAV capsid protein comprises the amino acid sequence of SEQ ID NO: 180.
2. The AAV vector of claim 1 , wherein the AAV vector comprises a cargo nucleic acid, wherein the cargo nucleic acid is encapsidated by the AAV capsid protein.
3. The AAV vector of claim 2 , wherein the cargo nucleic acid comprises at least one terminal repeat sequence.
4. The AAV vector of claim 3 , wherein the cargo nucleic acid comprises a heterologous nucleic acid sequence.
5. The AAV vector of claim 4 , wherein the cargo nucleic acid comprises inverted terminal repeat (ITR) sequences located 3′ and 5′ of the heterologous sequence.
6. The AAV vector of claim 4 , wherein the heterologous nucleic acid sequence encodes a polypeptide.
7. The AAV vector of claim 6 , wherein the polypeptide is a therapeutic polypeptide.
8. The AAV vector of claim 6 , wherein the polypeptide is an immunogenic polypeptide.
9. The AAV vector of claim 6 , wherein the polypeptide is nuclease.
10. The AAV vector of claim 9 , wherein the nuclease is a Cas9 nuclease, a Cas12(a) nuclease (Cpf1), a Cas12b nuclease, a Cas12c nuclease, a TrpB-like nuclease, a Cas13a nuclease (C2c2), a Cas13b nuclease, or a modified or truncated variant of any thereof.
11. The AAV vector of claim 4 , wherein the heterologous nucleic acid sequence encodes an untranslated RNA.
12. The AAV vector of claim 11 , wherein the untranslated RNA is a guide RNA.
13. The AAV vector of claim 11 , wherein the untranslated RNA is an antisense RNA, a ribozyme, or an interfering RNA.
14. A pharmaceutical composition comprising the AAV vector of claim 2 and a pharmaceutically acceptable carrier.
15. A method of introducing a cargo nucleic acid molecule into a cell, comprising contacting the cell with the AAV vector of claim 2 .
16. An adeno-associated virus (AAV) capsid protein comprising the amino acid sequence of SEQ ID NO: 180.
17. A nucleic acid comprising a nucleic acid sequence encoding the AAV capsid protein of claim 16 .
18. A cell comprising the nucleic acid of claim 17 .
19. An expression vector comprising a nucleic acid sequence encoding the AAV capsid protein of claim 16 .
20. A method of producing an adeno-associated virus (AAV) vector, the method comprising:
a. culturing a cell that comprises (i) the nucleic acid of claim 17 , (ii) a cargo nucleic acid comprising a 5′ inverted terminal repeat (ITR), a heterologous acid sequence, and a 3′ ITR, and (iii) AAV sequences sufficient for replication and encapsidation of nucleic acid, wherein the cell is cultured under conditions such that it produces the AAV vector; and
b. collecting the AAV vector from the cell.