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Gene knockdown in the mouse through RNAi

Item Type:Review
Title:Gene knockdown in the mouse through RNAi
Creators Name:Kleinhammer, A. and Wurst, W. and Kuehn, R.
Abstract:RNA interference (RNAi)-mediated gene knockdown has developed into a routine method to assess gene function in cultured mammalian cells in a fast and easy manner. For the use of RNAi in mice, short hairpin (sh) RNAs expressed from transgenic vectors are a fast alternative to conventional knockout approaches. We describe our strategy to elicit body-wide, cell type-specific, or inducible gene silencing in mice by control of shRNA expression through Cre recombinase or doxycycline. For reproducible expression of shRNAs, vectors are placed into the Rosa26 locus of ES cells by recombinase-mediated cassette exchange and transmitted through the germ line of chimeric mice. The site specific insertion of single copy shRNA vectors allows to expedite and reproducible production of knockdown mice and provides a simple approach to assess gene function in vivo.
Keywords:RNAi, Transgenic Mice, Rosa26, Cre/loxP, Doxycycline, RMCE, shRNA, Animals, Mice
Source:Methods in Enzymology
Series Name:Methods in Enzymology
Title of Book:Guide to Techniques in Mouse Development, Part B: Mouse Molecular Genetics
ISSN:0076-6879
ISBN:978-0-12-384880-2
Publisher:Elsevier / Academic Press (U.S.A.)
Volume:477
Page Range:387-414
Date:2010
Official Publication:https://doi.org/10.1016/S0076-6879(10)77020-8
PubMed:View item in PubMed

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