A Rapid Strategy to Detect the Recombined Allele in LSL-T beta RICA Transgenic Mice

作者：Vincent David F; Kaniewski Bastien; Powers Shannon E; Havenar Daughton Colin; Marie Julien C; Wotton David; Bartholin Laurent^*

来源：Genesis, 2010, 48(9): 559-562.

DOI：10.1002/dvg.20653

• TGF beta
• LSL
• PCR
• loxP
• CRE recombinase

We have previously generated a transgenic mouse strain (LSL-T beta RICA) containing a Cre-inducible constitutively active TGF beta 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 T beta RICA 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-T beta RICA 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-T beta RICA allele. genesis 48:559-562, 2010.

• 出版日期2010-9