Abstract
Lentiviral vectors have evolved over the last decade as powerful, reliable, and safe tools for stable gene transfer in a wide variety of mammalian cells. Contrary to other vectors derived from oncoretroviruses, they allow for stable gene delivery into most nondividing primary cells. In particular, lentivectors (LVs) derived from HIV-1 have gradually evolved to display many desirable features aimed at increasing both their safety and their versatility. This is why lentiviral vectors are becoming the most useful and promising tools for genetic engineering, to generate cells that can be used for research, diagnosis, and therapy. This chapter describes protocols and guidelines, for production and titration of LVs, which can be implemented in a research laboratory setting, with an emphasis on standardization in order to improve transposability of results between laboratories. We also discuss latest designs in LV technology.
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
References
Russell, S. J., and Cosset, F. L. (1999) Modifying the host range properties of retroviral vectors, J Gene Med 1, 300–311.
Naldini, L. et al. (1996) In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector, Science 272, 263–267.
Kobinger, G. P., Weiner, D. J., Yu, Q. C., and Wilson, J. M. (2001) Filovirus-pseudotyped lentiviral vector can efficiently and stably transduce airway epithelia in vivo, Nat Biotechnol 19, 225–230.
Duisit, G. et al. (2002) Five recombinant simian immunodeficiency virus pseudotypes lead to exclusive transduction of retinal pigmented epithelium in rat, Mol Ther 6, 446–454.
Stitz, J. et al. (2000) Lentiviral vectors pseudotyped with envelope glycoproteins derived from gibbon ape leukemia virus and murine leukemia virus 10A1, Virology 273, 16–20.
Hanawa, H. et al. (2002) Comparison of various envelope proteins for their ability to pseudotype lentiviral vectors and transduce primitive hematopoietic cells from human blood, Mol Ther 5, 242–251.
Sandrin, V. et al. (2002) Lentiviral vectors pseudotyped with a modified RD114 Âenvelope glycoprotein show increased stability in sera and augmented transduction of primary lymphocytes and CD34+ cells derived from human and nonhuman primates, Blood 100, 823–832.
Frecha, C. et al. (2008) Stable transduction of quiescent T cells without induction of cycle progression by a novel lentiviral vector pseudotyped with measles virus glycoproteins, Blood 112, 4843–4852.
Zufferey, R., Nagy, D., Mandel, R. J., Naldini, L., and Trono, D. (1997) Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo, Nat Biotechnol 15, 871–875.
Zufferey, R., Donello, J. E., Trono, D., and Hope, T. J. (1999) Woodchuck hepatitis virus posttranscriptional regulatory element enhances expression of transgenes delivered by retroviral vectors, J Virol 73, 2886–2892.
Follenzi, A., Ailles, L. E., Bakovic, S., Geuna, M., and Naldini, L. (2000) Gene transfer by lentiviral vectors is limited by nuclear translocation and rescued by HIV-1 pol sequences, Nat Genet 25, 217–222.
Zennou, V., Petit, C., Guetard, D., Nerhbass, U., Montagnier, L., and Charneau, P. (2000) HIV-1 genome nuclear import is mediated by a central DNA flap, Cell 101, 173–185.
Zufferey, R. et al. (1998) Self-inactivating lentivirus vector for safe and efficient In vivo gene delivery, J Virol 72, 9873–9880.
Dull, T. et al. (1998) A third-generation lentivirus vector with a conditional packaging system, J Virol 72, 8463–8471.
Gossen, M., Freundlieb, S., Bender, G., Muller, G., Hillen, W., and Bujard, H. (1995) Transcriptional activation by tetracyclines in mammalian cells, Science 268, 1766–1769.
Lois, C., Hong, E. J., Pease, S., Brown, E. J., and Baltimore, D. (2002) Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors, Science 295, 868–872.
Salmon, P., Oberholzer, J., Occhiodoro, T., Morel, P., Lou, J., and Trono, D. (2000) Reversible immortalization of human primary cells by lentivector- mediated transfer of specific genes, Mol Ther 2, 404–414.
Niwa, H., Yamamura, K., and Miyazaki, J. (1991) Efficient selection for high-expression transfectants with a novel eukaryotic vector, Gene 108, 193–199.
Mizushima, S., and Nagata, S. (1990) pEF-BOS, a powerful mammalian expression vector, Nucleic Acids Res 18, 5322.
Kostic, C. et al. (2003) Activity analysis of housekeeping promoters using self-inactivating lentiviral vector delivery into the mouse retina, Gene Ther 10, 818–821.
Escarpe, P. et al. (2003) Development of a sensitive assay for detection of replication-competent recombinant lentivirus in large-scale HIV-based vector preparations, Mol Ther 8, 332–341.
Frecha, C., Szecsi, J., Cosset, F. L., and Verhoeyen, E. (2008) Strategies for targeting lentiviral vectors, Curr Gene Ther 8, 449–460.
Mochizuki, H., Schwartz, J. P., Tanaka, K., Brady, R. O., and Reiser, J. (1998) High-titer human immunodeficiency virus type 1-based vector systems for gene delivery into nondividing cells, J Virol 72, 8873–8883.
Mazarakis, N. D. et al. (2001) Rabies virus glycoprotein pseudotyping of lentiviral vectors enables retrograde axonal transport and access to the nervous system after peripheral delivery, Hum Mol Genet 10, 2109–2121.
Wong, L. F. et al. (2004) Transduction Âpatterns of pseudotyped lentiviral vectors in the nervous system, Mol Ther 9, 101–111.
Azzouz, M. et al. (2004) Lentivector-mediated SMN replacement in a mouse model of spinal muscular atrophy, J Clin Invest 114, 1726–1731.
Watson, D. J., Kobinger, G. P., Passini, M. A., Wilson, J. M., and Wolfe, J. H. (2002) Targeted transduction patterns in the mouse brain by lentivirus vectors pseudotyped with VSV, Ebola, Mokola, LCMV, or MuLV envelope proteins, Mol Ther 5, 528–537.
Bemelmans, A. P. et al. (2005) Retinal cell type expression specificity of HIV-1-derived gene transfer vectors upon subretinal injection in the adult rat: influence of pseudotyping and promoter, J Gene Med 7, 1367–1374.
Miletic, H. et al. (2004) Selective transduction of malignant glioma by lentiviral vectors pseudotyped with lymphocytic choriomeningitis virus glycoproteins, Hum Gene Ther 15, 1091–1100.
Stein, C. S., Martins, I., and Davidson, B. L. (2005) The lymphocytic choriomeningitis virus envelope glycoprotein targets lentiviral gene transfer vector to neural progenitors in the murine brain, Mol Ther 11, 382–389.
Kang, Y. et al. (2002) In vivo gene transfer using a nonprimate lentiviral vector pseudotyped with Ross River Virus glycoproteins, J Virol 76, 9378–9388.
Medina, M. F. et al. (2003) Lentiviral vectors pseudotyped with minimal filovirus envelopes increased gene transfer in murine lung, Mol Ther 8, 777–789.
Silvertown, J. D., Walia, J. S., Summerlee, A. J., and Medin, J. A. (2006) Functional expression of mouse relaxin and mouse relaxin-3 in the lung from an Ebola virus glycoprotein-pseudotyped lentivirus via tracheal delivery, Endocrinology 147, 3797–3808.
Hachiya, A. et al. (2007) Gene transfer in human skin with different pseudotyped HIV-based vectors, Gene Ther 14, 648–656.
Kang, Y. et al. (2005) Persistent expression of factor VIII in vivo following nonprimate lentiviral gene transfer, Blood 106, 1552–1558.
Bartosch, B., Dubuisson, J., and Cosset, F. L. (2003) Infectious hepatitis C virus pseudo-particles containing functional E1-E2 envelope protein complexes, J Exp Med 197, 633–642.
Kowolik, C. M., and Yee, J. K. (2002) Preferential transduction of human hepatocytes with lentiviral vectors pseudotyped by Sendai virus F protein, Mol Ther 5, 762–769.
Christodoulopoulos, I., and Cannon, P. M. (2001) Sequences in the cytoplasmic tail of the gibbon ape leukemia virus envelope protein that prevent its incorporation into lentivirus vectors, J Virol 75, 4129–4138.
Frecha, C. et al. (2009) Efficient and stable transduction of resting B lymphocytes and primary chronic lymphocyte leukemia cells using measles virus gp displaying lentiviral vectors, Blood 114, 3173–3180.
Acknowledgements
We thank Ophélie Cherpin and David Suter for their help in the construction and design of Gateway® lentivectors.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC 2011
About this protocol
Cite this protocol
Giry-Laterrière, M., Verhoeyen, E., Salmon, P. (2011). Lentiviral Vectors. In: Merten, OW., Al-Rubeai, M. (eds) Viral Vectors for Gene Therapy. Methods in Molecular Biology, vol 737. Humana Press. https://doi.org/10.1007/978-1-61779-095-9_8
Download citation
DOI: https://doi.org/10.1007/978-1-61779-095-9_8
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-094-2
Online ISBN: 978-1-61779-095-9
eBook Packages: Springer Protocols