A rapid strategy to detect the recombined allele in LSL-TβRICA transgenic mice.
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.
Vincent
David F
DF
INSERM, U590, Lyon, France.
Kaniewski
Bastien
B
Powers
Shannon E
SE
Havenar-Daughton
Colin
C
Marie
Julien C
JC
Wotton
David
D
Bartholin
Laurent
L
D000818:Animals; D018388:Codon, Terminator / Q000235:genetics; D003433:Crosses, Genetic; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005838:Genotype; D019426:Integrases / Q000235:genetics; D008099:Liver / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D010179:Pancreas / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics*; D011995:Recombination, Genetic / Q000235:genetics*; D055748:SOXB1 Transcription Factors / Q000235:genetics; D013601:T-Lymphocytes / Q000378:metabolism; D019076:Transgenes / Q000235:genetics*
D018388:Codon, Terminator; D017931:DNA Primers; D018125:Receptors, Transforming Growth Factor beta; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; D017346:Protein Serine-Threonine Kinases; D000077293:Receptor, Transforming Growth Factor-beta Type I; C045073:Cre recombinase; D019426:Integrases