Conditional and inducible transgene expression in mice through the combinatorial use of Cre-mediated recombination and tetracycline induction

Nucleic Acids Res. 2005 Mar 22;33(5):e51. doi: 10.1093/nar/gni051.

Abstract

Here we describe a triple transgenic mouse system, which combines the tissue specificity of any Cre-transgenic line with the inducibility of the reverse tetracycline transactivator (rtTA)/tetracycline-responsive element (tet-O)-driven transgenes. To ensure reliable rtTA expression in a broad range of cell types, we have targeted the rtTA transgene into the ROSA26 locus. The rtTA expression, however, is conditional to a Cre recombinase-mediated excision of a STOP region from the ROSA26 locus. We demonstrate the utility of this technology through the inducible expression of the vascular endothelial growth factor (VEGF-A) during embryonic development and postnatally in adult mice. Our results of adult induction recapitulate several different hepatic and immune cell pathological phenotypes associated with increased systemic VEGF-A protein levels. This system will be useful for studying genes in which temporal control of expression is necessary for the discovery of the full spectrum of functions. The presented approach abrogates the need to generate tissue-specific rtTA transgenes for tissues where well-characterized Cre lines already exist.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Doxycycline / pharmacology*
  • Embryo, Mammalian / cytology
  • Gene Expression Regulation*
  • Gene Targeting
  • Integrases / genetics
  • Integrases / metabolism*
  • Mice
  • Mice, Transgenic / genetics*
  • Mice, Transgenic / metabolism
  • Proteins / genetics
  • RNA, Untranslated
  • Recombination, Genetic*
  • Stem Cells / metabolism
  • Trans-Activators / genetics
  • Trans-Activators / metabolism
  • Transcriptional Activation
  • Transgenes
  • Vascular Endothelial Growth Factor A / biosynthesis
  • Vascular Endothelial Growth Factor A / genetics
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*

Substances

  • Gt(ROSA)26Sor non-coding RNA, mouse
  • Proteins
  • RNA, Untranslated
  • Trans-Activators
  • Vascular Endothelial Growth Factor A
  • Viral Proteins
  • Cre recombinase
  • Integrases
  • Doxycycline