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Production of high-titer helper-free retroviruses by transient transfection

Pear, Warren S. and Nolan, Garry P. and Scott, Martin L. and Baltimore, David (1993) Production of high-titer helper-free retroviruses by transient transfection. Proceedings of the National Academy of Sciences of the United States of America, 90 (18). pp. 8392-8396. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:20120308-093523110

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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.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1073/pnas.90.18.8392DOIUNSPECIFIED
http://www.pnas.org/content/90/18/8392PublisherUNSPECIFIED
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.
Funders:
Funding AgencyGrant Number
Howard Hughes Postdoctoral Fellowship for PhysiciansUNSPECIFIED
Leukemia Society of AmericaUNSPECIFIED
NIH CA51462
Subject Keywords:retroviral packaging cells; gene therapy
Record Number:CaltechAUTHORS:20120308-093523110
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20120308-093523110
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:29641
Collection:CaltechAUTHORS
Deposited By: Jason Perez
Deposited On:12 Mar 2012 20:49
Last Modified:25 Mar 2014 06:35

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