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

Prem K Premsrirut, Lukas E Dow, Sang Yong Kim, Matthew Camiolo, Colin D Malone, Cornelius Miething, Claudio Scuoppo, Johannes Zuber, Ross Alexander Dickins, Scott C Kogan, Kenneth R Shroyer, Raffaella Sordella, Gregory J Hannon, Scott W Lowe

Research output: Contribution to journalArticleResearchpeer-review

204 Citations (Scopus)

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, p16INK4a, p19ARF 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 p19ARF 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.
Original languageEnglish
Pages (from-to)145 - 158
Number of pages14
JournalCell
Volume145
DOIs
Publication statusPublished - 2011
Externally publishedYes

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