A Rapid and Scalable System for Studying Gene Function in Mice Using Conditional RNA Interference
- Premsrirut, PK; Dow, LE; Kim, SY; Camiolo, M; Malone, CD; Miething, C; Scuoppo, C; Zuber, J; Dickins, RA; Kogan, SC; Shroyer, KR; Sordella, R; Hannon, GJ; Lowe, SW;
Publication Year 2011-04-01, Volume 145, Issue #1, Page 145-158
- Journal Title
- Publication Type
- Journal Article
- 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.
- CELL PRESS
- EMBRYONIC STEM-CELLS; TRANSGENIC MICE; LUNG ADENOCARCINOMAS; MAMMALIAN-CELLS; IN-VIVO; K-RAS; MOUSE; EXPRESSION; DIFFERENTIATION; APC
- Publisher's Version
- Rights Notice
- Refer to copyright notice on published article.
Creation Date: 2011-04-01 12:00:00Last Modified: 0001-01-01 12:00:00