ADENO-ASSOCIATED VIRUS (AAV) CLADES, SEQUENCES, VECTORS CONTAINING SAME, AND USES THEREFOR

• US 20190054188A1
• Filed: 10/02/2018
• Published: 02/21/2019
• Est. Priority Date: 09/30/2003
• Status: Active Grant

• Current Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Wilson, James M., Gao, Guangping, Alvira, Mauricio R., Vandenberghe, Luc H.
• Granted Patent: US 10,695,441 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2750/14121 Viruses as such, e.g. new isolates, mutants or their genomic sequences
• CPC Class Codes: C12N 2750/14122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2750/14142 virus or viral particle as vehicle, e.g. encapsulating small organic molecule
• CPC Class Codes: C12N 2750/14143 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2750/14151 Methods of production or purification of viral material
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination
• CPC Class Codes: C12N 2830/48 regulating transport or export of RNA, e.g. RRE, PRE, WPRE, CTE
• CPC Class Codes: C12N 2830/85 mammalian
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00
• CPC Class Codes: C12N 9/644 Coagulation factor IXa (3.4.21.22)
• CPC Class Codes: C12Y 304/21022 Coagulation factor IXa (3.4.21.22)

Abstract

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.

Claims

1. A recombinant nucleic acid molecule:

(a) encoding an AAV9 vp1 capsid protein having a sequence comprising amino acids 1 to 736 of SEQ ID NO;

123;

or

(b) comprising nucleotides 1 to 2208 of SEQ ID NO;

3, or a nucleotide sequence at least 99% identical to nucleotides 1 to 2208 of SEQ ID NO;

3,

wherein the recombinant nucleic acid molecule does not contain an AAV inverted terminal repeat.

2. The recombinant nucleic acid molecule according to claim 1, which further comprises a functional rep gene.

3. The recombinant nucleic acid molecule according to claim 1, wherein said recombinant nucleic acid molecule comprises a sequence selected from:

vp1, nt 1 to 2208;

vp2, nt 412 to 2208;

or

vp3, nt 607 to 2208 of SEQ ID NO;

3.

4. The recombinant nucleic acid molecule according to claim 1, wherein said recombinant nucleic acid molecule is a plasmid.

5. A host cell transfected with the recombinant nucleic acid molecule according to claim 1.

6. The host cell according to claim 5, which further comprises a functional rep gene, a minigene comprising AAV inverted terminal repeats (ITRs) and a transgene, and sufficient helper functions to permit packaging of the minigene into the AAV capsid.

7. The host cell according to claim 6, which further comprises a nucleic acid sequence

(a) encoding an AAV9 vp2 capsid protein having a sequence comprising amino acids 138 to 736 of SEQ ID NO:

123;

or

(b) comprising nucleotides 412 to 2208 of SEQ ID NO;

3, or a nucleotide sequence at least 99% identical to nucleotides 412 to 2208 of SEQ ID NO;

3.

8. The host cell according to claim 6, which further comprises a nucleic acid sequence

(a) encoding an AAV9 vp3 capsid protein having a sequence comprising amino acids 203 to 736 of SEQ ID NO:

123;

or

(b) comprising nucleotides 607 to 2208 of SEQ ID NO;

3, or a nucleotide sequence at least 99% identical to nucleotides 607 to 2208 of SEQ ID NO;

3.

9. A recombinant AAV comprising an AAV8 capsid produced by the host cell of claim 6.

10. A recombinant AAV comprising an AAV8 capsid protein encoded by the recombinant nucleic acid molecule according to claim 1.

11. A composition comprising the recombinant AAV according to claim 10 and a pharmaceutically acceptable carrier.

12. A recombinant nucleic acid molecule:

(a) encoding an AAV9 vp2 capsid protein having a sequence comprising amino acids 138 to 736 of SEQ ID NO;

123;

or

(b) comprising nucleotides 412 to 2208 of SEQ ID NO;

3, or a nucleotide sequence at least 99% identical to nucleotides 412 to 2208 of SEQ ID NO;

3,

wherein the recombinant nucleic acid molecule does not contain an AAV inverted terminal repeat.

13. The recombinant nucleic acid molecule according to claim 12, which further comprises a functional rep gene.

14. The recombinant nucleic acid molecule according to claim 12, wherein said recombinant nucleic acid molecule is a plasmid.

15. A host cell transfected with the recombinant nucleic acid molecule according to claim 12.

16. The host cell according to claim 15, which further comprises a functional rep gene, a minigene comprising AAV inverted terminal repeats (ITRs) and a transgene, and sufficient helper functions to permit packaging of the minigene into the AAV capsid.

17. A recombinant AAV comprising an AAV8 capsid produced by the host cell of claim 16.

18. A recombinant AAV comprising an AAV8 capsid protein encoded by the recombinant nucleic acid molecule according to claim 12.

19. A composition comprising the recombinant AAV according to claim 18 and a pharmaceutically acceptable carrier.

20. A recombinant nucleic acid molecule:

(a) encoding an AAV9 vp3 capsid protein having a sequence comprising amino acids 203 to 736 of SEQ ID NO;

123;

or

(b) comprising nucleotides 607 to 2208 of SEQ ID NO;

3, or a nucleotide sequence at least 99% identical to nucleotides 607 to 2208 of SEQ ID NO;

3,

wherein the recombinant nucleic acid molecule does not contain an AAV inverted terminal repeat.

21. The recombinant nucleic acid molecule according to claim 20, which further comprises a functional rep gene.

22. The recombinant nucleic acid molecule according to claim 20, wherein said recombinant nucleic acid molecule is a plasmid.

23. A host cell transfected with the recombinant nucleic acid molecule according to claim 20.

24. The host cell according to claim 23, which further comprises a functional rep gene, a minigene comprising AAV inverted terminal repeats (ITRs) and a transgene, and sufficient helper functions to permit packaging of the minigene into the AAV capsid.

25. A recombinant AAV comprising an AAV8 capsid produced by the host cell of claim 24.

26. A recombinant AAV comprising an AAV8 capsid protein encoded by the recombinant nucleic acid molecule according to claim 20.

27. A composition comprising the recombinant AAV according to claim 26 and a pharmaceutically acceptable carrier.

28. An AAV9 vector comprising an AAV9 capsid, said capsid having packaged therein a nucleic acid molecule comprising AAV inverted terminal repeat sequences and a gene encoding a product, which gene is operably linked to regulatory sequences which direct expression of the gene product, wherein said AAV9 capsid is produced from expression in an AAV packaging cell comprising a AAV9 vp1 nucleotide sequence which is at least about 90% identical to nucleotides 1 to 2208 of SEQ ID NO:

3 and which encodes amino acids 1 to 736 of SEQ ID NO;

123.