SpringerProtocols: Abstract: Generation of High-Titer, Helper-Free Retroviruses by Transient Transfection

Generation of High-Titer, Helper-Free Retroviruses by Transient Transfection

By: Warren S. Pear^{2 3}, Martin L. Scott², Garry P. Nolan⁴

Abstract

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Retroviral gene transfer is presently one of the most powerful techniques for introducing stably heritable genetic material into mammalian cells (reviewed in ref. 1). One serious drawback of this technique, however, has been the difficulty in readily producing high-titer recombinant retroviruses. For many applications, such as infecting rare target cells or the majority of cells in tissue culture, the recombinant virus titer must be at least 10⁶ infectious units/mL. Although one can usually obtain high-titer mixtures of recombinant and replication-competent retroviruses in a relatively short time, many applications such as cell marking studies or studying genes in vivo demand freedom from replication-competent virus.

Affiliation(s): (2) Department of Biology, Massachusetts Institute of Technology, Cambridge, MA
(3) Department of Pathology, University of Pennsylvania Medical Center, Philadelphia, PA
(4) Department of Pharmacology, Stanford University, Stanford, CA

Book Title: Gene Therapy Protocols

Series: Methods in Molecular Medicine | Volume: 7 | Pub. Date: Oct-01-1996 | Page Range: 41-57 | DOI: 10.1385/0-89603-484-4:41

Subject: Genetics/Genomics

References

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1.	Mulligan, R. C. (1993) The basic science of gene therapy. Science 260, 926–932.

2.	Mann, R., Mulligan, R. C, and Baltimore, D. (1983) Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus. Cell 33, 153–159.

3.	Watanabe, S. and Temin, H. M. (1983) Construction of a helper cell line for avian reticuloendotheliosis virus cloning vectors. Mol Cell. Biol. 3, 2241–2249.

4.	Miller, A. D. (1990) Retrovirus packaging cells. Human Gene Ther. 1, 5–14.

5.	Danos, O. (1991) Construction of retroviral packaging cell lines, in Methods in Molecular Biology, vol. 8 (Collins, M. ed.), Humana, Clifton, NJ, pp. 17–26.

6.	Landau, N. R. and Littman, D. R. (1992) Packaging system for rapid generation of murine leukemia virus vectors with variable tropism. J. Viral 66, 5110–5113.

7.	Pear, W., Nolan, G., Scott, M., and Baltimore, D. (1993) Production of high-titer helperfree retroviruses by transient transfection. Proc. Natl. Acad. Sci USA 90, 8392–8396.

8.	Finer, M. H., Dull, T. J., Qin, L., Farson, D., and Roberts, M. R. (1994) Kat: a high-efficiency retroviral transduction system for primary human T lymphocytes. Blood 83, 43–50.

9.	Scadden, D. T., Fuller, B., and Cunningham, J. M. (1990) Human cells Infected with retrovirus vectors acquire an endogenous murine provirus. J Virol 64, 424–427.

10.	Graham, F. L., Smiley, J., Russell, W. C., and Naish, R. (1977) Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J. Gen Virol. 36, 59–72.

11.	DuBridge, R. B., Tang, P., Hsia, H C., Phaik-Mooi, L, Miller, J H, and Calos, M. P. (1987) Analysis of mutation in human cells by using an Epstein-Barr virus shuttle system Mol Cell. Biol. 7, 379–387.

12.	Danos, O. and Mulligan, R. C. (1988) Safe and efficient generation of recombi-nant retroviruses with amphotropic and ecotropic host ranges. Proc. Natl Acad Sci. USA 85, 6460–6464.

13.	Bender, M A., Palmer, T D, Gelinas, R. E, and Miller, A. D (1987) Evidence that the packaging signal of Moloney murine leukemia virus extends into the gag region. J Virol. 61, 1639–1646.

14.	Daley, G. and Baltimore, D. (1988) Transformation of an interleukin 3-dependent hematopoietic cell line by the chronic myelogenous leukemia-specific P210 bcr / abl protein. Proc. Natl Acad Sci USA 85, 9312–9316.

15.	Morgenstern, J. P. and Land, H. (1990) Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line. Nucleic Acids Res 18, 3587–3596.

16.	Dranoff, G., Jaffee, E., Lazenby, A., Golumbek, P., Levitsky, H., Brose, K, Jackson, V., Hamada, H., Pardoll, D., and Mulligan, R C. (1993) Vaccination with irradiated tumor cells engineered to secrete murine granulocyte-macrophage colony-stimulating factor stimulates potent, specific, and long-lasting anti-tumor immunity. Proc Natl. Acad. Sci USA 90, 3539–3543.

17.	Shoemaker, C, Hoffmann, J., Goff, S. P, and Baltimore, D. (1981) Intramolecular integration within moloney murine leukemia virus DNA. J Viral 40, 164–172.

18.	Bernard, H. U., Krammer, G., and Rowekamp, W. G. (1985) Construction of a fusion gene that confers resistance against hygromycin B to mammalian cells in culture. Exp. Cell Res 158, 237–243.

19.	Goff, S., Traktman, P., and Baltimore, D. (1981) Isolation and properties of Moloney murine leukemia virus mutants: use of a rapid assay for release of virion reverse transcriptase .J Virol. 38, 239–248.

20.	Jasin, M. and Berg, P. (1988) Homologous integration in mammalian cells with-out target gene selection. Genes Dev 2, 1353–1363.

21.	Burns, J. C., Friedmann, T., Driever, W., Burrascano, M., and Yee, J. K. (1993) Vesicular stomatitis virus G glycoprotein pseudotyped retroviral vectors: concentration to a very high titer and efficient gene transfer into mammalian and nonmammalian cells. Proc Natl. Acad. Sci. USA 90, 8033–8037.

22.	Mulligan, R. C. and Berg, P. (1981) Selection for animal cells that express the escherichia coli gene coding for xanthine-guanine phosphoribosyltransferase. Proc. Natl. Acad. Sci. USA 78, 2072–2076.

23.	Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A., and Strubl, K. (1989) Current Protocols in Molecular Biology, Wiley, New York.

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