Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses
Patent
1993-01-21
1995-07-25
Mosher, Mary E.
Chemistry: molecular biology and microbiology
Treatment of micro-organisms or enzymes with electrical or...
Modification of viruses
4352402, 4352351, 4353201, 435 914, 536 2372, C12N 1500, C12N 510, C12N 701, C12N 1535
Patent
active
054361460
ABSTRACT:
The present invention relates to a method for producing helper-free stocks of recombinant adeno-associated virus (rAAV) which can be used to efficiently and stably transduce foreign genes into host cells or organisms. The method comprises the cotransfection of eukaryotic cells with rAAV and with helper AAV DNA in the presence of helper virus (e.g. adenovirus or herpesvirus) such that the helper AAV DNA is not associated with virion formation. The crux of the invention lies in the inability of the helper AAV DNA to recombine with rAAV vector, thereby preventing the generation of wild-type virus. In a specific embodiment of the invention, the vector comprises a recombinant AAV genome containing only the terminal regions of the AAV chromosome bracketing a non-viral gene, and the helper AAV DNA comprises a recombinant AAV genome containing that part of the AAV genome which is not present in the vector, and in which the AAV terminal regions are replaced by adenovirus sequences. In a further embodiment of the invention, cell lines are created which incorporate helper AAV DNA which can directly produce substantially pure recombinant AAV virus. The pure stocks of recombinant AAV produced according to the invention provide an AAV viral expression vector system with increased yield of recombinant virus, improved efficiency, higher definition, and greater safety than presently used systems.
REFERENCES:
patent: 4797368 (1989-01-01), Carter et al.
Labow, M. A. et al. 1988, J. Virol. vol. 62 pp. 1705-1712.
DeLaMaza, L. M. et al. 1980 J. Biol. Chem. vol. 255 pp. 3194-3203.
Lebkowski, J. et al. 1988. Molecular and Cellular Biology, vol. 8 pp. 3988-3996.
Samulski, R. J. et al. 1987. Journal of Virology, vol. 61, pp. 3096-3101.
McLaughlin, S. K. et al. 1988. Journal of Virology, vol. 62, pp. 1963-1973.
Miller, A. D. et al. Somatic Cell and Molecular Genetics, vol. 12, pp. 175-183.
Bosselman, R. A. et al. 1987. Molecular and Cellular Biology, vol. 7, pp. 1979-1806.
Laface et al., 1988, Virology 162:483-486.
Ohi et al., 1988, J. Cell Biol. 107:304A.
Wondisford et al., 1988, Mol. Endocrinol. 2:32-39.
Hermonat and Muzyczka, 1984, Proc. Natl. Acad. Sci. U.S.A. 81:6466-6470.
Senapathy et al., 1984, J. Mol. Biol. 179:1-20.
Tratschin et al., 1984, Mol. Cell. Biol. 4:2072-2081.
Laughlin et al., 1983, Gene 23:65-73.
Samulski et al., 1983, Cell 33:135-143.
Samulski et al., 1982, Proc. Natl. Acad. Sci. U.S.A. 79:2077-2081.
Chang Long-Sheng
Samulski Richard J.
Shenk Thomas E.
Mosher Mary E.
The Trustees of Princeton University
LandOfFree
Helper-free stocks of recombinant adeno-associated virus vectors does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Helper-free stocks of recombinant adeno-associated virus vectors, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Helper-free stocks of recombinant adeno-associated virus vectors will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-739095