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Conditional Gene Expression and Knockdown Using Lentivirus Vectors Encoding shRNA

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Gene Therapy Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 434))

Summary

Drug-inducible systems allowing the control of transgene expression and knockdown in mammalian cells are invaluable tools for genetic research, and could also play important roles in translational research or gene therapy. We and others have developed a lentivector-based, conditional gene expression system for drug-controllable expression of transgenes and small hairpin RNAs (shRNAs). This system is highly robust and versatile, governing tightly controlled expression of transgenes and endogenous cellular genes (through shRNAs) in various primary and established cell lines in vitro, as well as in vivo in the central nervous system or in human cancer cells xenotransplanted into nude mice. The goal of this article is to provide a concise methodology for construction and manipulation of this conditional lentiviral-based system, and quantitative analyses of drug-inducible transgene expression and gene knockdown both in vitro and in vivo.

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Acknowledgments

Authors thank Maciej Wiznerowicz for critical reading of this chapter.

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© 2008 Humana Press, a part of Springer Science+Business Media, LLC

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Szulc, J., Aebischer, P. (2008). Conditional Gene Expression and Knockdown Using Lentivirus Vectors Encoding shRNA. In: Le Doux, J.M. (eds) Gene Therapy Protocols. Methods in Molecular Biology™, vol 434. Humana Press. https://doi.org/10.1007/978-1-60327-248-3_18

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  • DOI: https://doi.org/10.1007/978-1-60327-248-3_18

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60327-247-6

  • Online ISBN: 978-1-60327-248-3

  • eBook Packages: Springer Protocols

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