title: A rapid and scalable system for studying gene function in mice using conditional RNA interference.
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:
authors: Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
authors: Prem K
authors: PK
authors: Premsrirut
authors: Lukas E
authors: LE
authors: Dow
authors: Sang Yong
authors: SY
authors: Kim
authors: Matthew
authors: M
authors: Camiolo
authors: Colin D
authors: CD
authors: Malone
authors: Cornelius
authors: C
authors: Miething
authors: Claudio
authors: C
authors: Scuoppo
authors: Johannes
authors: J
authors: Zuber
authors: Ross A
authors: RA
authors: Dickins
authors: Scott C
authors: SC
authors: Kogan
authors: Kenneth R
authors: KR
authors: Shroyer
authors: Raffaella
authors: R
authors: Sordella
authors: Gregory J
authors: GJ
authors: Hannon
authors: Scott W
authors: SW
authors: Lowe
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000628:therapy; D000818:Animals; D053595:Embryonic Stem Cells / Q000378:metabolism; D055785:Gene Knockdown Techniques / Q000191:economics / Q000379:methods*; D008175:Lung Neoplasms / Q000235:genetics / Q000628:therapy; D051379:Mice; D008822:Mice, Transgenic; D035683:MicroRNAs / Q000235:genetics; D054218:Precursor T-Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics / Q000628:therapy; D034622:RNA Interference; D012323:RNA Processing, Post-Transcriptional; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction; D051153:Wnt Proteins / Q000378:metabolism
chemical_list: D035683:MicroRNAs; D034741:RNA, Small Interfering; D051153:Wnt Proteins