Abstract
Retroviral vectors derived from murine leukemia retrovirus (MuLV) have been widely used for efficient gene transfer to achieve long-term expression of a chosen therapeutic gene in mammLian cells (1). Disadvantages of this vector are the instability and low viral titers generated from packaging cells, low efficiency of gene transfer into human cells, especially in vivo, and the requirement for dividing cells. Some authors have attempted to increase the transduction efficiency by using strategies like low-speed centrifugation of viral supernatant with cells, multiple viral exposures (2), or increasing viral titers by ultracentrifugation (3); they were able to produce an average transduction efficiency of 10–60%. However, all such improvements in transduction efficiency require additional procedures, which are practically inefficient.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Miller A. D., Miller D. G., Garcia J. V., and Lynch C. M. (1993) Use of retroviral vectors for gene transfer and expression. Methods Enzymol. 217, 581–599.
Inaba M., Toninelli E., Vanmeter G., Bender J. R., and Conte M. S. (1998) Retroviral gene transfer: effects on endothelial cell phenotype. J. Surg. Res. 78, 31–36.
Zelenock J. A., Welling T. H., Sarkar R., Gordon D. G., and Messina L. M. (1997) Improved retroviral transduction efficiency of vascular cells in vitro and in vivo during clinically relevant incubation periods using centrifugation to increase viral titers. J Vasc. Surg. 26, 119–127.
Friedmann T. and Yee J. K. (1995) Pseudotyped retroviral vectors for studies of human gene therapy. NatMed. 1, 275–277.
Schnell M. J., Buonocore L., Kretzschmar E., Johnson E., and Rose J. K. (1996) Foreign glycoproteins expressed from recombinant vesicular stomatitis viruses are incorporated efficiently into virus particles. Proc. Natl. Acad. Sci. USA 93, 11359–11365.
Burns J. C., Friedmann T., Driever W., Burrascano M., and Yee J. K. (1993) Vesicular stomatitis virus G glycoprotein pseudotyped retroviral vectors: concentration to very high titer and efficient gene transfer into mammalian and nonmammalian cells [see comments]. Proc. Natl. Acad. Sci. USA 90, 8033–8037.
Gallardo H. F., Tan C., Ory D., and Sadelain M. (1997) Recombinant retroviruses pseudotyped with the vesicular stomatitis virus G glycoprotein mediate both stable gene transfer and pseudotransduction in human peripheral blood lymphocytes. Blood 90, 952–957.
An D. S., Koyanagi Y., Zhao J. Q., et al. (1997) High-efficiency transduction of human lymphoid progenitor cells and expression in differentiated T cells. J. Virol. 71, 1397–1404.
Sharma S., Cantwell M., Kipps T. J., and Friedmann T. (1996) Efficient infection of a human T-cell line and of human primary peripheral blood leukocytes with a pseudotyped retrovirus vector. Proc. Natl. Acad. Sci. USA 93, 11842–11847.
Liu M. L., Winther B. L., and Kay M. A. (1996) Pseudotransduction of hepatocytes by using concentrated pseudotyped vesicular stomatitis virus G glycoprotein (VSV-G)-Moloney murine leukemia virus-derived retrovirus vectors: comparison of VSV-G and amphotropic vectors for hepatic gene transfer. J. Virol. 70, 2497–2502.
Yu H., Eton D., Wang Y., et al. (1999) High efficiency in vitro gene transfer into vascular tissues using a pseudotyped retroviral vector without pseudotransduction. Gene Ther. 6, 1876–1883.
Emi N., Friedmann T., and Yee J. K. (1991) Pseudotype formation of murine leukemia virus with the G protein of vesicular stomatitis virus. J. Virol. 65, 1202–1207.
Schlegel R., Tralka T. S., Willingham M. C., and Pastan I. (1983) Inhibition of VSV binding and infectivity by phosphatidylserine: is phosphatidylserine a VSVbinding site? Cell 32, 639–646.
Mastromarino P., Conti C., Goldoni P., Hauttecoeur B., and Orsi N. (1987) Characterization of membrane components of the erythrocyte involved in vesicular stomatitis virus attachment and fusion at acidic pH. J. Gen. Virol. 68, 2359–2369.
Conti C., Mastromarino P., Ciuffarella M. G., and Orsi N. (1988) Characterization of rat brain cellular membrane components acting as receptors for vesicular stomatitis virus. Brief report. Arch. Virol. 99, 261–269.
Chen S. T., Iida A., Guo L., Friedmann T., and Yee J. K. (1996) Generation of packaging cell lines for pseudotyped retroviral vectors of the G protein of vesicular stomatitis virus by using a modified tetracycline inducible system. Proc. Natl. Acad. Sci.USA 93, 10057–10062.
Ory D. S., Neugeboren B. A., and Mulligan R. C. (1996) A stable human-de-rived packaging cell line for production of high titer retrovirus/vesicular stomatitis virus G pseudotypes. Proc. Natl. Acad. Sci. USA 93, 11400–11406.
Arai T., Matsumoto K., Saitoh K., et al. (1998) A new system for stringent, high-titer vesicular stomatitis virus G protein-pseudotyped retrovirus vector induction by introduction of Cre recombinase into stable prepackaging cell lines. J. Virol. 72, 1115–1121.
Miller D. G., Adam M. A., and Miller A. D. (1990) Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection [published erratum appears in Mol Cell Biol (1992) 12,433. Mol. Cell. Biol. 10, 4239–4242.
Pear W. S., Nolan G. P., Scott M. L., and Baltimore D. (1993) Production of high-titer helper-free retroviruses by transient transfection. Proc. Natl. Acad. Sci.USA 90, 8392–8396.
Han J. Y., Cannon P. M., Lai K. M., Zhao Y., Eiden M. V., and Anderson W. F. (1997) Identification of envelope protein residues required for the expended host range of 10A1 murine leukemia birus. J. Virol. 71, 8103–8108.
Soneoka Y., Cannon P. M., Ramsdale E. E., et al. (1995) A transient threeplasmid expression system for the production of high titer retroviral vectors. Nucleic Acids Res. 23, 628–633.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2002 Humana Press Inc.
About this protocol
Cite this protocol
Yu, H. (2002). Preparation of Pseudotyped Retroviral Vector. In: Morgan, J.R. (eds) Gene Therapy Protocols. Methods in Molecular Medicine, vol 69. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-141-8:149
Download citation
DOI: https://doi.org/10.1385/1-59259-141-8:149
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-723-6
Online ISBN: 978-1-59259-141-1
eBook Packages: Springer Protocols