| Literature DB >> 21458673 |
Prem K Premsrirut1, Lukas E Dow, Sang Yong Kim, Matthew Camiolo, Colin D Malone, Cornelius Miething, Claudio Scuoppo, Johannes Zuber, Ross A Dickins, Scott C Kogan, Kenneth R Shroyer, Raffaella Sordella, Gregory J Hannon, Scott W Lowe.
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:Entities:
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Year: 2011 PMID： 21458673 PMCID： PMC3244080 DOI： 10.1016/j.cell.2011.03.012
Source DB: PubMed Journal: Cell ISSN： 0092-8674 Impact factor: 41.582