Abstract
Ecotropic and amphotropic retrovirus packaging Cell lines have been constructed in which the helper virus genome have been separated onto two plasmids, and the ψ packaging signal and 3’ LTR have been removed. The gag and pol genes on one plasmid and the env gene on another plasmid were transfected into NIH 3T3 Cells. Packaging Cell lines produced by these transfected genes released titers of replication-defective retroviral vectors which were comparable to titers produced by packaging Cell lines containing the helper virus genome on one plasmid. There has been no evidence of recombination events between the ecotropic helper virus plasmids and the vector virus plasmid that would result in the generation of intact replication-competent virus. These results suggest that a packaging Cell line containing gag, pol and env on different plasmids is efficient and safe for use in retroviral gene gransfer.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Cone, R.D., A. Webber-Benarous, D. Baorto, and R.C. Mulligan. 1987. Regulated expression of a complete human ß-globin gene encoded by a transmissible retrovirus vector. Mol. Cell. Biol. 7: 887–897.
Anderson, W.F. 1984. Prospects for human gene therapy. Science 226: 401–409.
Neel, B.B., W.S. Hayward, H.L. Robinson, J. Fang, and S. M. Astrin. 1981. Avian leukosis virus-induced tumors have common proviral integration sites and synthesize discrete new RNAs: Oncogenesis by promoter insertion. Cell 23: 323–334.
Varmus, H.E., N. Quintrell, and S. Ortiz. 1981. Retroviruses as mutagenss Insertion and exision of a nontransforming provirus alter expression of a resident transforming provirus. Cell 25: 23–36.
Cone, R.D., and R.C. Mulligan. 1984. High efficiency gene transfer into mammalian Cells: Generation of helper-free retrovirus with broad mammalian host range. Proc. Natl. Acad. Sci. 81: 6349–6353.
Mann, R., F.C. Mulligan, and D. Baltimore. 1983. Construction of a retrovirus packaging mutant and its use to produce helper- free defective retrovirus. Cell 33: 153–159.
Miller, A.D., M.-F. Law, and I.M. Verma. 1985. Generation of helper-free amphotropic retroviruses that transduce a dominant- acting methotrexate-resistant dihydrofolate reductase gene. Mol. Cell Biol. 5: 431–437.
Sorge, J., D. Wright, V.D. Erdman, and A. Cutting. 1984. Amphotropic retrovirus system for human Cell gene transfer. Mol. Cell. Biol. 4: 1730–1737.
Watanabe, S., and H.M. Temin. 1983. Construction of a helper Cell line for Avian reticuloendotheliosis virus cloning factors. Mol. Cell. Biol. 3: 2241–2249.
Hock, R.A., and A.D. Miller. 1986. Retrovirus-mediated transfer and expression of drug-resistant genes in human haematopoietic progenitor Cells. Nature 320: 257–277.
Miller, A.D., and C. Buttimore. 1986. Redesign of retrovirus packaging Cell lines to avoid recombination leading to helper virus production. Mol. Cell. Biol. 6: 2895–2902.
Mulligan, R.C., and P. Berg. 1980. Expression of a bacterial gene in mammalian Cells. Science 209: 175–183.
Potter, H., L.W. Weir, and P. Leder. 1984. Enhancer-dependent expression of human K immunoglobulin genes introduced into mouse pre-B lymphocytes by electroporation. Proc. Natl. Acad. Sci. 81: 7161–7165.
Goff, S.P., P. Traktman, and D. Baltimore. 1981. Isolation and properties of Molony murine leukemia virus mutants: Use of a rapid assay for release of virion reverse transcriptase. J. Virol. 38: 239–248.
Schwartzberg, P., J. Colicelli, and S. Goff. 1983. Deletion mutants of Molony murine leukemia virus which lack glycosylated gag protein are replication competent. J. Virol. 46: 538–546.
Keller, G., C. Paige, E. Gilboa, and E.F. Wagner. 1985. Expression of a foreign gene in myeloid and lymphoid Cells derived from multipotent haematopoietic precursors. Nature 318: 149–154.
Murphy, A.J. 1987. Molecular techniques for the isolation of transcriptional transacting genes. Doctoral Thesis, Columbia University 133–139.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1988 Plenum Press, New York
About this chapter
Cite this chapter
Markowitz, D., Goff, S., Bank, A. (1988). Construction of a Safe and Efficient Retrovirus Packaging Cell Line. In: Tavassoli, M., Zanjani, E.D., Ascensao, J.L., Abraham, N.G., Levine, A.S. (eds) Molecular Biology of Hemopoiesis. Advances in Experimental Medicine and Biology, vol 34. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-5571-7_6
Download citation
DOI: https://doi.org/10.1007/978-1-4684-5571-7_6
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4684-5573-1
Online ISBN: 978-1-4684-5571-7
eBook Packages: Springer Book Archive