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Cell. 2011 Apr 1;145(1):145-58. doi: 10.1016/j.cell.2011.03.012.

A rapid and scalable system for studying gene function in mice using conditional RNA interference.

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  • 1Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.

Abstract

RNA interference is a powerful tool for studying gene function, however, the reproducible generation of RNAi transgenic mice remains a significant limitation. By combining optimized fluorescence-coupled miR30-based shRNAs with high efficiency ES cell targeting, we developed a fast, scalable pipeline for the production of shRNA transgenic mice. Using this system, we generated eight tet-regulated shRNA transgenic lines targeting Firefly and Renilla luciferases, Oct4 and tumor suppressors p53, p16(INK4a), p19(ARF) and APC and demonstrate potent gene silencing and GFP-tracked knockdown in a broad range of tissues in vivo. Further, using an shRNA targeting APC, we illustrate how this approach can identify predicted phenotypes and also unknown functions for a well-studied gene. In addition, through regulated gene silencing we validate APC/Wnt and p19(ARF) as potential therapeutic targets in T cell acute lymphoblastic leukemia/lymphoma and lung adenocarcinoma, respectively. This system provides a cost-effective and scalable platform for the production of RNAi transgenic mice targeting any mammalian gene. PAPERCLIP:

Copyright © 2011 Elsevier Inc. All rights reserved.

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PMID:
21458673
[PubMed - indexed for MEDLINE]
PMCID:
PMC3244080
Free PMC Article
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