A pipeline for the generation of shRNA transgenic mice
Details
Publication Year 2012-02, Volume 7, Issue #2, Page 374-393
Journal Title
NATURE PROTOCOLS
Publication Type
Journal Article
Abstract
RNA interference (RNAi) is an extremely effective tool for studying gene function in almost all metazoan and eukaryotic model systems. RNAi in mice, through the expression of short hairpin RNAs (shRNAs), offers something not easily achieved with traditional genetic approaches-inducible and reversible gene silencing. However, technical variability associated with the production of shRNA transgenic strains has so far limited their widespread use. Here we describe a pipeline for the generation of miR30-based shRNA transgenic mice that enables efficient and consistent targeting of doxycycline-regulated, fluorescence-linked shRNAs to the Col1a1 locus. Notably, the protocol details crucial steps in the design and testing of miR30-based shRNAs to maximize the potential for developing effective transgenic strains. In all, this 14-week procedure provides a fast and cost-effective way for any laboratory to investigate gene function in vivo in the mouse.
Publisher
NATURE PUBLISHING GROUP
Keywords
Cell biology ; Genetic modification ; Model organisms
WEHI Research Division(s)
Molecular Medicine
Rights Notice
© 2012 Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.


Creation Date: 2012-02-01 12:00:00
Last Modified: 0001-01-01 12:00:00
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