A pipeline for the generation of shRNA transgenic mice. Academic Article uri icon

Overview

abstract

  • RNA interference (RNAi) is an extremely effective tool for studying gene function in almost all metazoan and eukaryotic model systems. RNAi in mice, through the expression of short hairpin RNAs (shRNAs), offers something not easily achieved with traditional genetic approaches-inducible and reversible gene silencing. However, technical variability associated with the production of shRNA transgenic strains has so far limited their widespread use. Here we describe a pipeline for the generation of miR30-based shRNA transgenic mice that enables efficient and consistent targeting of doxycycline-regulated, fluorescence-linked shRNAs to the Col1a1 locus. Notably, the protocol details crucial steps in the design and testing of miR30-based shRNAs to maximize the potential for developing effective transgenic strains. In all, this 14-week procedure provides a fast and cost-effective way for any laboratory to investigate gene function in vivo in the mouse.

authors

  • Dow, Lukas Edward
  • Premsrirut, Prem K
  • Zuber, Johannes
  • Fellmann, Christof
  • McJunkin, Katherine
  • Miething, Cornelius
  • Park, Youngkyu
  • Dickins, Ross A
  • Hannon, Gregory J
  • Lowe, Scott W

publication date

  • February 2, 2012

Research

keywords

  • Genetic Engineering
  • Mice, Transgenic
  • RNA Interference
  • RNA, Small Interfering

Identity

PubMed Central ID

  • PMC3724521

Scopus Document Identifier

  • 84858382422

Digital Object Identifier (DOI)

  • 10.1038/nprot.2011.446

PubMed ID

  • 22301776

Additional Document Info

volume

  • 7

issue

  • 2