Claims
- 1. A nucleic acid molecule which encodes one or more AAV helper functions, said nucleic acid molecule comprising:
an AAV rep coding region; an AAV cap coding region; and a nucleotide sequence comprising a modified AAV p5 promoter region, such that the modified AAV p5 promoter region no longer functions in transcription initiation.
- 2. An AAV helper function vector comprising the nucleic acid molecule of claim 1.
- 3. The AAV helper function vector of claim 2, wherein the vector is a plasmid.
- 4. The AAV helper function vector of claim 2, further comprising one or more accessory function genes.
- 5. A method of producing recombinant AAV (rAAV) virions comprising:
(a) introducing an AAV vector into a suitable host cell; (b) introducing the AAV helper function vector of claim 2 in to the host cell; (c) expressing accessory functions in the host cell; and (d) culturing the host cell to produce rAAV virions.
- 6. A recombinant AAV (rAAV) virion produced by the method of claim 5.
- 7. A host cell comprising the nucleic acid molecule of claim 1.
- 8. The host cell of claim 7 further comprising accessory function genes.
- 9. A nucleic acid molecule which encodes one or more AAV helper functions, said nucleic acid molecule comprising:
an AAV rep coding region; an AAV cap coding region; and a nucleotide sequence comprising a modified AAV p5 promoter region, wherein the modified promoter region lacks an intact TATA box.
- 10. An AAV helper function vector comprising the nucleic acid molecule of claim 9.
- 11. The AAV helper function vector of claim 10, wherein the vector is a plasmid.
- 12. The AAV helper function vector of claim 10, further comprising one or more accessory function genes.
- 13. A method of producing recombinant AAV (rAAV) virions comprising:
(a) introducing an AAV vector into a suitable host cell; (b) introducing the AAV helper function vector of claim 10 into the host cell; (c) expressing accessory functions in the host cell; and (d) culturing the host cell to produce rAAV virions.
- 14. A recombinant AAV (rAAV) virion produced by the method of claim 13.
- 15. A host cell comprising the nucleic acid molecule of claim 9.
- 16. The host cell of claim 15 further comprising accessory function genes.
- 17. The nucleic acid molecule of claim 9, wherein the nucleotide sequence is arranged in the nucleic acid molecule such that the modified AAV p5 promoter region is situated 3′ relative to the rep coding region.
- 18. An AAV helper function vector comprising the nucleic acid molecule of claim 17.
- 19. The AAV helper function vector of claim 18, wherein the vector is a plasmid.
- 20. The AAV helper function vector of claim 18, further comprising one or more accessory function genes.
- 21. A method of producing recombinant AAV (rAAV) virions comprising:
(a) introducing an AAV vector into a suitable host cell; (b) introducing the AAV helper function vector of claim 18 into the host cell; (c) expressing accessory functions in the host cell; and (d) culturing the host cell to produce rAAV virions.
- 22. A recombinant AAV (rAAV) virion produced by the method of claim 21.
- 23. A host cell comprising the nucleic acid molecule of claim 17.
- 24. The host cell of claim 23 further comprising accessory function genes.
- 25. The nucleic acid molecule of claim 9, wherein the nucleotide sequence is arranged in the nucleic acid molecule such that the modified AAV p5 promoter region is situated 3′ relative to the cap coding region.
1. RELATED APPLICATIONS
[0001] This application is a continuation of copending U.S. patent application Ser. No. 09/143,270 of Peter Colosi filed Aug. 28, 1998 and entitled “High Efficiency Wild-Type-Free AAV Helper Functions,” which is a continuation-in-part of U.S. patent application Ser. No. 09/107,708 of Georges Natsoulis, Peter Colosi, and Gary Kurtzman filed Jun. 30, 1998 and entitled “High Efficiency AAV Helper Functions” which is a continuation-in-part of U.S. patent application Ser. No. 08/688,648 of Georges Natsoulis filed Jul. 29, 1996 and entitled “High Efficiency Helper System for AAV Vector Production,” which is a continuation-in-part of U.S. patent application Ser. No. 08/510,790 of Georges Natsoulis filed Aug. 3, 1995 and entitled “High Efficiency Helper System for AAV Vector Production,” which issued as U.S. Pat. No. 5,622,856 on Apr. 22, 1997. These prior applications are incorporated herein by reference.
Continuations (2)
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Number |
Date |
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Parent |
09450083 |
Nov 1999 |
US |
Child |
10074302 |
Feb 2002 |
US |
Parent |
09143270 |
Aug 1998 |
US |
Child |
09450083 |
Nov 1999 |
US |
Continuation in Parts (3)
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Number |
Date |
Country |
Parent |
09107708 |
Jun 1998 |
US |
Child |
09143270 |
Aug 1998 |
US |
Parent |
08688648 |
Jul 1996 |
US |
Child |
09107708 |
Jun 1998 |
US |
Parent |
08510790 |
Aug 1995 |
US |
Child |
08688648 |
Jul 1996 |
US |