Production of high-titer helper-free retroviruses by transient transfection
Abstract
The generation of high-titer, helper-free retroviruses by transient transfection has been achieved by using the highly transfectable 293T cell line into which are stably introduced constructs that express retroviral packaging functions. The resulting ecotropic virus packaging cell line BOSC 23 produces infectious retrovirus at > 10^(6) infectious units/ml of supernatant within 72 hr after CaPO_(4)-mediated transfection. A stringent assay for replication-competent virus showed that no helper virus was present. The system can produce high titers of retroviral vectors expressing genes that are extremely difficult to propagate at high titer in stable producer lines. This method should facilitate and extend the use of helper-free retroviral gene transfer, as well as be useful for gene therapy.
Additional Information
© 1993 National Academy of Sciences. Contributed by David Baltimore, May 27, 1993. We thank Dr. Richard Mulligan and members of his laboratory for many helpful discussions and for providing the CRIP, CRE, and MFG constructs. We thank Dr. Steve Gofffor critical review of this manuscript. We thank Dr. C. Cepko for pBND 8 and Dr. M. Jasin for pGPT2E. W.S.P. was supported by a Howard Hughes Postdoctoral Fellowship for Physicians. G.P.N. was supported by a grant from the Leukemia Society of America. This work was supported by National Institutes of Health Grant CA51462 to D.B.Attached Files
Published - PEApnas93.pdf
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Additional details
- PMCID
- PMC47362
- Eprint ID
- 29641
- Resolver ID
- CaltechAUTHORS:20120308-093523110
- Howard Hughes Medical Institute (HHMI)
- Leukemia Society of America
- CA51462
- NIH
- Created
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2012-03-12Created from EPrint's datestamp field
- Updated
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2021-11-09Created from EPrint's last_modified field