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

Overview

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

  • Premsrirut, Prem K
  • Dow, Lukas Edward
  • Kim, Sang Yong
  • Camiolo, Matthew
  • Malone, Colin D
  • Miething, Cornelius
  • Scuoppo, Claudio
  • Zuber, Johannes
  • Dickins, Ross A
  • Kogan, Scott C
  • Shroyer, Kenneth R
  • Sordella, Raffaella
  • Hannon, Gregory J
  • Lowe, Scott W

publication date

  • April 1, 2011

Research

keywords

  • Gene Knockdown Techniques
  • RNA Interference

Identity

PubMed Central ID

  • PMC3244080

Scopus Document Identifier

  • 79952696452

Digital Object Identifier (DOI)

  • 10.1016/j.cell.2011.03.012

PubMed ID

  • 21458673

Additional Document Info

volume

  • 145

issue

  • 1