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Journal of Virology, September 1998, p. 7024-7031, Vol. 72, No. 9
Markey Molecular Medicine Center and
Department of Medicine, University of Washington, Seattle,
Washington 98195
Received 29 January 1998/Accepted 21 May 1998
Although vectors based on adeno-associated virus (AAV) offer
several unique advantages, their usage has been hampered by the difficulties encountered in vector production. In this report, we
describe a new AAV packaging system based on inducible
amplification of integrated helper and vector constructs containing the
simian virus 40 (SV40) replication origin. The packaging and producer cell lines developed express SV40 T antigen under the control of the reverse tetracycline transactivator system, which allows inducible amplification of chromosomal loci linked to the SV40 origin. Culturing these cells in the presence of doxycycline followed by adenovirus infection resulted in helper and vector gene
amplification as well as higher vector titers. Clonal producer cell
lines generated vector titers that were 10 times higher than those
obtained by standard methods, with approximately 104
vector particles produced per cell. These stocks were free of detectable replication-competent virus. The lack of a transfection step
combined with the reproducibility of stable producer lines makes this packaging method ideally suited for the
large-scale production of vector stocks for human gene therapy.
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Packaging Cells Based on Inducible Gene
Amplification for the Production of Adeno-Associated Virus
Vectors
*
Corresponding author. Mailing address: Department of
Medicine, Box 357720, University of Washington, Seattle, WA 98195. Phone: (206) 616-4562. Fax: (206) 616-8298. E-mail:
drussell{at}u.washington.edu.
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