Summary
RNA interference (RNAi) has recently emerged as a reliable tool for studying the effects of knocking down or ablating the expression of specific genes. It is hoped that progress made in the laboratory toward in vitro down regulation of gene expression may be carried over into the clinic for treatment of diseases in which the expression of a specific gene is associated with initiation or progression of that disease. Such is the case with telomerase, an exciting drug target that has been the focus of numerous investigations with a wide variety of inhibitors. This chapter describes the use of retrovirally introduced short-hairpin RNA as an effector of stable, long-term RNAi in human cells.
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References
Devroe, E. and Silver, P. A. (2002) Retrovirus-delivered siRNA. BMC Bio-technol. 2, 15.
Zhou, H., Xia, X. G., and Xu, Z. (2005) An RNA polymerase II construct synthesizes short-hairpin RNA with a quantitative indicator and mediates highly efficient RNAi. Nucleic. Acids Res. 33, e62.
Brummelkamp, T. R., Bernards, R., and Agami, R. (2002) A system for stable expression of short interfering RNAs in mammalian cells. Science 296, 550–553.
Brummelkamp, T. R., Bernards, R., and Agami, R. (2002) Stable suppression of tumorigenicity by virus-mediated RNA interference. Cancer Cell. 2, 243–247.
Sui, G., Soohoo, C., Affar el, B., Gay, F., Shi, Y., and Forrester, W. C. (2002) A DNA vector-based RNAi technology to suppress gene expression in mammalian cells. Proc. Natl. Acad. Sci. U. S. A. 99, 5515–5520.
Stewart, S. A., Dykxhoorn, D. M., Palliser, D., Mizuno, H., Yu, E. Y., and An, D. S. (2003) Lentivirus-delivered stable gene silencing by RNAi in primary cells. RNA 9, 493–501.
Liu, C. M., Liu, D. P., Dong, W. J., and Liang, C. C. (2004) Retrovirus vector-mediated stable gene silencing in human cell. Biochem. Biophys. Res. Commun. 313, 716–720.
Paddison, P. J., Caudy, A. A., Bernstein, E., Hannon, G. J., and Conklin, D. S. (2002) Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells. Genes Dev. 16, 948–958.
Elbashir, S. M., Harborth, J., Lendeckel, W., Yalcin, A., Weber, K., and Tuschl, T. (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411, 494–498.
Elbashir, S. M., Harborth, J., Weber, K., and Tuschl, T. (2002) Analysis of gene function in somatic mammalian cells using small interfering RNAs. Methods 26, 199–213.
Masutomi, K., Yu, E. Y., Khurts, S., Ben-Porath, I., Currier, J. L., and Metz, G. B. (2003) Telomerase maintains telomere structure in normal human cells. Cell 114, 241–253.
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© 2007 Humana Press Inc.
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Cunningham, A.P., Andrews, L.G., Tollefsbol, T.O. (2007). Retrovirus-Mediated RNA Interference. In: Andrews, L.G., Tollefsbol, T.O. (eds) Telomerase Inhibition. Methods in Molecular Biology™, vol 405. Humana Press. https://doi.org/10.1007/978-1-60327-070-0_5
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DOI: https://doi.org/10.1007/978-1-60327-070-0_5
Publisher Name: Humana Press
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Online ISBN: 978-1-60327-070-0
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