Adeno-associated virus (AAV) serotype 8 sequences, vectors containing same, and uses therefor

1. An isolated adeno-associated virus (AAV) comprising an AAV8 capsid having an amino acid sequence of amino acids 1 to 737 of SEQ ID NO:2 or a sequence which is at least 95% identical therewith.

2. The isolated AAV according to claim 1 , wherein said AAV further comprises a minigene having AAV inverted terminal repeats and a heterologous gene operably linked to regulatory sequences which direct its expression in a host cell.

3. A host cell transfected with an adeno-associated virus according to claim 1 in culture.

4. A composition comprising an AAV according to claim 1 and a physiologically compatible carrier.

5. A method of delivering a transgene to a cell, said method comprising the step of contacting the cell with an AAV according to claim 1 , wherein said AAV is a recombinant AAV (rAAV) comprising a transgene.

6. An isolated capsid protein comprising an AAV8 protein selected from the group consisting of:

vp1capsid protein having the sequence of amino acids (aa) 1 to 737 of SEQ ID NO:2;

vp2 capsid protein having the sequence of aa 138 to 737 of SEQ ID NO:2; and

vp3 capsid protein having the sequence of aa 203 to 737 of SEQ ID NO:2.

7. A recombinant adeno-associated viral (AAV) vector comprising an AAV8 capsid and a nucleotide sequence encoding a Factor VIII transgene operably linked to expression control elements.

8. The recombinant AAV vector according to claim 7 , wherein the Factor VIII transgene is selected from the group consisting of:

(a) the light chain of Factor VIII;

(b) the heavy chain of Factor VIII; and

(c) a B-domain mutant of Factor VIII.

9. A method of delivering a nucleotide sequence encoding a Factor VIII protein to a mammal, said method comprising the step of administering a recombinant AAV vector according to claim 7 to a mammal under conditions that result in the expression of the Factor VIII protein at a level that provides a therapeutic effect in said mammal.

10. The method according to claim 9 further comprising administering a first rAAV comprising an AAV8 capsid which comprises a nucleotide sequence encoding the light chain of Factor VIII operably linked to expression control elements and administering a second rAAV8 which comprises a nucleotide sequence encoding the heavy chain of Factor VIII operably linked to expression control elements.

11. The method according to claim 9 , wherein said expression control elements comprise a tissue-specific promoter.

12. The method according to claim 9 , where said expression control elements comprise a human growth hormone polyadenylation sequence.

13. The method according to claim 11 , wherein the tissue-specific promoter is a liver-specific promoter.

14. A method of treating hemophilia in a mammal comprising: administering a rAAV having a serotype 8 capsid and a nucleic acid sequence encoding a Factor VIII transciene to a mammal under conditions that result in the expression of the Factor VIII protein at a level that provides a therapeutic effect in said mammal.

15. A viral vector comprising an AAV8 capsid and a transgene encoding a product under control of regulatory elements that direct expression of the product in a host cell.

16. The viral vector according to claim 15 , wherein said AAV8 capsid is characterized by having a sequence selected from the group consisting of amino acid residues 1 to 737 of SEQ ID NO:2 or an amino acid sequence at least 95% identical thereto.

17. An isolated AAV8 rep protein having the amino acid sequence of amino acids 1 to 625 of SEQ ID NO:3.

18. An isolated AAV8 rep protein fragment selected from the group consisting of amino acid (aa) 1 to 102; aa 103 to 140; aa 141 to 173; aa 174 to 226; aa 227 to 275; aa 276 to 374; aa 375 to 383; aa 384 to 446; aa 447 to 542; aa 543 to 555; and aa 556 to 625, of SEQ ID NO: 3.