Authors:
Prem K. Premsrirut, 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, and Scott W. Lowe
Summary:
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.
Source:
Cell; Vol. 145, Issue 1, 145-158 (04/01/11)