Authors:
PK Premsrirut, LE Dow, SY Kim, M Camiolo, CD Malone, C Miething, C Scuoppo, J Zuber, RA Dickins, SC Kogan, KR Shroyer, R Sordella, GJ Hannon, SW Lowe
Journal name: 
Cell
Citation info: 
145(1):145-158
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:
DOI: 
http://doi.org/10.1016/j.cell.2011.03.012
Research group: 
Hannon Group
E-pub date: 
01 Apr 2011