Method of detecting and/or identifying adeno-associated virus (AAV) sequences and isolating novel sequences identified thereby

Patent metadata

Field	Value
Publication number	US-11041171-B2
Application number	US-202016682712-A
Country	US
Kind code	B2
Filing date	Nov 13, 2019
Priority date	Nov 13, 2001
Publication date	Jun 22, 2021
Grant date	Jun 22, 2021

Abstract

Adeno-associated virus rh.20 sequences, vectors containing same, and methods of use are provided.

First claim

The invention claimed is:

1. A method of generating a recombinant adeno-associated virus (AAV) comprising culturing a host cell containing: (a) a molecule encoding the AAV vp1 capsid protein having a sequence of amino acids 1 to 738 of SEQ ID NO: 85, or a sequence which is at least 95% identical to the full length of amino acids 1 to 738 of SEQ ID NO: 85; (b) a functional rep gene; (c) a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product operably linked to sequences which direct expression of the product in a host cell; and (d) sufficient helper functions to permit packaging of the minigene into the AAV capsid protein under conditions which permit packaging of the minigene into the AAV capsid.

2. The method of claim 1 , wherein the sequence of the vp1 protein is at least 97% identical to the full length of amino acids 1 to 738 of SEQ ID NO: 85.

3. The method of claim 1 , wherein the sequence of the vp1 protein is at least 99% identical to the full-length of amino acids 1 to 738 of SEQ ID NO: 85.

4. The method of claim 1 , wherein the sequence of the vp1 protein is the full-length of amino acids 1 to 738 of SEQ ID NO: 85.

5. The method of claim 1 , wherein the gene product is alpha-1 antitrypsin, Factor VIII, Factor IX, ornithine transcarbamylase, glucose-6-phosphatase, phenylalanine hydroxylase, argininosuccinate synthetase, β-glucuronidase (GUSB), or a dystrophin protein.

6. The method of claim 1 , wherein the molecule is a nucleic acid molecule comprising nucleotides 844 to 3057 of SEQ ID NO: 27, or a nucleotide sequence at least 99% identical to nucleotides 844 to 3057 of SEQ ID NO: 27.

7. The method of claim 1 , wherein the rep gene is from AAV2.

8. A method of generating a recombinant adeno-associated virus (AAV) comprising culturing a host cell containing: (a) a molecule encoding the AAV vp2 capsid protein having a sequence of amino acids 138 to 738 of SEQ ID NO: 85, or a sequence which is at least 95% identical to the full length of amino acids 138 to 738 of SEQ ID NO: 85; (b) a functional rep gene; (c) a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product operably linked to sequences which direct expression of the product in a host cell; and (d) sufficient helper functions to permit packaging of the minigene into the AAV capsid protein under conditions which permit packaging of the minigene into the AAV capsid.

9. The method of claim 8 , wherein the sequence of the vp2 protein is at least 97% identical to the full length of amino acids 138 to 738 of SEQ ID NO: 85.

10. The method of claim 8 , wherein the sequence of the vp2 protein is at least 99% identical to the full-length of amino acids 138 to 738 of SEQ ID NO: 85.

11. The method of claim 8 , wherein the sequence of the vp2 protein is the full-length of amino acids 138 to 738 of SEQ ID NO: 85.

12. The method of claim 8 , wherein the gene product is alpha-1 antitrypsin, Factor VIII, Factor IX, ornithine transcarbamylase, glucose-6-phosphatase, phenylalanine hydroxylase, argininosuccinate synthetase, β-glucuronidase (GUSB), or a dystrophin protein.

13. The method of claim 8 , wherein the molecule is a nucleic acid molecule comprising nucleotides 1255 to 3057 of SEQ ID NO: 27, or a nucleotide sequence at least 99% identical to nucleotides 1255 to 3057 of SEQ ID NO: 27.

14. The method of claim 8 , wherein the rep gene is from AAV2.

15. A method of generating a recombinant adeno-associated virus (AAV) comprising culturing a host cell containing: (a) a molecule encoding the AAV vp3 capsid protein having a sequence of amino acids 204 to 738 of SEQ ID NO: 85, or a sequence which is at least 95% identical to the full length of amino acids 204 to 738 of SEQ ID NO: 85; (b) a functional rep gene; (c) a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product operably linked to sequences which direct expression of the product in a host cell; and (d) sufficient helper functions to permit packaging of the minigene into the AAV capsid protein under conditions which permit packaging of the minigene into the AAV capsid.

16. The method of claim 15 , wherein the sequence of the vp3 protein is at least 97% identical to the full length of amino acids 204 to 738 of SEQ ID NO: 85.

17. The method of claim 15 , wherein the sequence of the vp3 protein is at least 99% identical to the full-length of amino acids 204 to 738 of SEQ ID NO: 85.

18. The method of claim 15 , wherein the sequence of the vp3 protein is the full-length of amino acids 204 to 738 of SEQ ID NO: 85.

19. The method of claim 15 , wherein the gene product is alpha-1 antitrypsin, Factor VIII, Factor IX, ornithine transcarbamylase, glucose-6-phosphatase, phenylalanine hydroxylase, argininosuccinate synthetase, β-glucuronidase (GUSB), or a dystrophin protein.

20. The method of claim 15 , wherein the molecule is a nucleic acid molecule comprising nucleotides 1453 to 3057 of SEQ ID NO: 27, or a nucleotide sequence at least 99% identical to nucleotides 1453 to 3057 of SEQ ID NO: 27.

21. The method of claim 15 , wherein the rep gene is from AAV2.

Assignees

• Univ Pennsylvania

Inventors

• Gao Guangping
• Wilson James M
• Alvira Mauricio R

Classifications

• A61K38/177

  Receptors; Cell surface antigens; Cell surface determinants · CPC title

• C12N2830/85

  mammalian · CPC title

• C12N2830/48

  regulating transport or export of RNA, e.g. RRE, PRE, WPRE, CTE · CPC title

• A61P11/00

  Drugs for disorders of the respiratory system · CPC title

• A61K38/4846

  Factor VII (3.4.21.21); Factor IX (3.4.21.22); Factor Xa (3.4.21.6); Factor XI (3.4.21.27); Factor XII (3.4.21.38) · CPC title