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

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:

Pubmed ID: 21458673


  • 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 Data

April 1, 2011

Associated Grants

  • Agency: NCI NIH HHS, Id: P01 CA013106
  • Agency: NCI NIH HHS, Id: P01 CA013106-40
  • Agency: NCI NIH HHS, Id: P30 CA008748
  • Agency: NIGMS NIH HHS, Id: R01 GM062534
  • Agency: Howard Hughes Medical Institute, Id:
  • Agency: Howard Hughes Medical Institute, Id:

Mesh Terms

  • Adenocarcinoma
  • Animals
  • Embryonic Stem Cells
  • Gene Knockdown Techniques
  • Lung Neoplasms
  • Mice
  • Mice, Transgenic
  • MicroRNAs
  • Precursor T-Cell Lymphoblastic Leukemia-Lymphoma
  • RNA Interference
  • RNA Processing, Post-Transcriptional
  • RNA, Small Interfering
  • Signal Transduction
  • Wnt Proteins