A rapid strategy to detect the recombined allele in LSL-TβRICA transgenic mice

Genesis. 2010 Sep;48(9):559-62. doi: 10.1002/dvg.20653.

Abstract

We have previously generated a transgenic mouse strain (LSL-TβRI(CA)) containing a Cre-inducible constitutively active TGFβ type I receptor (Bartholin et al., 2008, Genesis 46: 724-731). Transgene expression depends on the excision of a floxed-transcriptional STOP (LSL, Lox-STOP-Lox) located upstream the TβRI(CA) coding sequence. To evaluate the correct excision of the STOP signal in the presence of Cre-recombinase, we developed a rapid screening based on an original PCR genotyping strategy. More precisely, we designed a set of primers flanking the LSL containing region. The size of the amplified products will differ according to recombination status of the LSL-TβRI(CA) allele. Indeed, the size of the STOP containing PCR product is 1.93 kb, but is reduced to 0.35 kb when the STOP signal is removed after Cre-mediated recombination. We validated excision in several compartments, including pancreas, liver, T lymphocytes, and embryos using different Cre expressing transgenic mouse strains. This represents a simple and efficient way of monitoring the tissue specific recombination of the LSL-TβRI(CA) allele.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Codon, Terminator / genetics
  • Crosses, Genetic
  • DNA Primers / genetics
  • Female
  • Genotype
  • Integrases / genetics
  • Liver / metabolism
  • Mice
  • Mice, Transgenic
  • Pancreas / metabolism
  • Polymerase Chain Reaction / methods*
  • Protein-Serine-Threonine Kinases / genetics*
  • Receptor, Transforming Growth Factor-beta Type I
  • Receptors, Transforming Growth Factor beta / genetics*
  • Recombination, Genetic / genetics*
  • SOXB1 Transcription Factors / genetics
  • T-Lymphocytes / metabolism
  • Transgenes / genetics*

Substances

  • Codon, Terminator
  • DNA Primers
  • Receptors, Transforming Growth Factor beta
  • SOXB1 Transcription Factors
  • Sox2 protein, mouse
  • Protein-Serine-Threonine Kinases
  • Receptor, Transforming Growth Factor-beta Type I
  • Cre recombinase
  • Integrases