Adeno-associated virus (AAV) glades, sequences, vectors containing same, and uses therefor
Sequences of novel adeno-associated virus capsids and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. AAV-mediated delivery of therapeutic and immunogenic genes using the vectors of the invention is also provided.
1. A recombinant adeno-associated virus (AAV) comprising an AAVhu.37 capsid comprising vp1, vp2 and vp3 proteins, said vp1 having the amino acid sequence of amino acids 1 to 738 of SEQ ID NO:88, said AAV having packaged in the capsid a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product, the non-AAV nucleic acid sequence operably linked to sequences which direct expression of the product in a host cell.
2. The recombinant AAV according to claim 1 wherein the AAV ITR is from AAV2.
3. The recombinant AAV according to claim 1 , wherein the gene product is a protein selected from a Factor VIII, a Factor IX, an erythropoietin and an ornithine transcarbamylase (OTC).
4. The recombinant AAV according to claim 1 , wherein the gene product is an antibody.
5. A composition comprising a pharmaceutically acceptable carrier and at least a recombinant AAV comprising an AAVhu.37 capsid comprising vp1, vp2 and vp3 proteins, said vp1 having the amino acid sequence of amino acids 1 to 738 of SEQ ID NO:88, said AAV having packaged in the capsid a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product, the non-AAV nucleic acid sequence operably linked to sequences which direct expression of the product in a host cell.
6. A recombinant AAV comprising an AAVhu.37 capsid comprising vp1, vp2 and vp3 proteins, said vp3 having the amino acid sequence of amino acids 204 to 738 of SEQ ID NO: 88, said AAV having packaged in the capsid a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product, the non-AAV nucleic acid sequence operably linked to sequences which direct expression of the product in a host cell.
7. The recombinant AAV according to claim 6 wherein the AAV ITR is from AAV2.
8. The recombinant AAV according to claim 6 , wherein the gene product is a protein selected from a Factor VIII, a Factor IX, an erythropoietin and an ornithine transcarbamylase (OTC).
9. The recombinant AAV according to claim 6 , wherein the gene product is an antibody.
10. The recombinant AAV according to claim 6 , wherein the vp1 protein has the amino acid sequence of amino acids 1 to 738 of SEQ ID NO:88.
11. The recombinant AAV according to claim 6 , wherein the vp2 protein has the amino acid sequence of amino acids 138 to 738 of SEQ ID NO:88.
12. The recombinant AAV according to claim 6 , wherein the vp1 protein has the amino acid sequence of amino acids 1 to 738 of SEQ ID NO:88, the vp2 protein has the amino acid sequence of amino acids 138 to 738 of SEQ ID NO:88 and the vp3 protein has the amino acid sequence of amino acids 204 to 738 of SEQ ID NO:88.
13. A composition comprising a pharmaceutically acceptable carrier and at least a recombinant AAV comprising an AAVhu.37 capsid comprising vp1, vp2 and vp3 proteins, said vp2 having the amino acid sequence of amino acids 138 to 738 of SEQ ID NO:88, said AAV having packaged in the capsid a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product, the non-AAV nucleic acid sequence operably linked to sequences which direct expression of the product in a host cell.
14. A recombinant AAV comprising an AAVhu.37 capsid comprising vp1, vp2 and vp3 proteins, said vp2 having the amino acid sequence of amino acids 138 to 738 of SEQ ID NO: 88, said AAV having packaged in the capsid a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product, the non-AAV nucleic acid sequence operably linked to sequences which direct expression thereof in a host cell.
15. The recombinant AAV according to claim 14 wherein the AAV ITR is from AAV2.
16. The recombinant AAV according to claim 14 , wherein the gene product is a protein selected from a Factor VIII, a Factor IX, an erythropoietin and an ornithine transcarbamylase (OTC).
17. The recombinant AAV according to claim 14 , wherein the gene product is an antibody.
18. A composition comprising a pharmaceutically acceptable carrier and at least a recombinant AAV comprising an AAVhu.37 capsid comprising vp1, vp2 and vp3 proteins, said vp3 having the amino acid sequence of amino acids 204 to 738 of SEQ ID NO:88, said AAV having packaged in the capsid a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product, the non-AAV nucleic acid sequence operably linked to sequences which direct expression of the product in a host cell.
19. A method of generating a recombinant AAV comprising an AAV hu.37 capsid comprising the steps of culturing a host cell containing: (a) a nucleic acid sequence encoding the AAVhu.37 vp1 capsid protein having the amino acid sequence of amino acids 1 to 738 of SEQ ID NO: 88; (b) a sequence encoding an AAV rep; (c) a nucleic acid sequence comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product, the non-AAV nucleic acid sequence operably linked to sequences which direct expression of the product in a host cell and (d) sufficient helper functions for packaging the AAV.