B6(Cg)-S1pr2^tm1.1Thla/J

Stock number: 035004
Product name: S1pr2^f
Research areas: Cancer Research, Cardiovascular Research, Research Tools

Description

S1pr2^f mice possess loxP sites flanking exon 2 of the sphingosine-1-phosphate receptor 2 (S1pr2) gene. These floxed mice may be useful when studying vascular development and maturation, as well as tumor growth.

Detailed description

S1pr2^f mice possess loxP sites flanking exon 2 of the sphingosine-1-phosphate receptor 2 (S1pr2) gene. This gene encodes a receptor for sphingosine 1-phosphate (S1P), a lysophospholipid found in blood and lymph that regulates cell survival, migration, immune cell trafficking, angiogenesis, and vascular barrier function. Mice that are homozygous for this floxed allele are viable and fertile. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exon 2 deleted in the Cre-expressing tissues.

When crossed to Cdh5-CreER mice, resulting S1pr2 ECKO mice have endothelial-specific deletion of S1pr2 after induction with tamoxifen. ECKO mice shows no vascular phenotype, yet they display smaller tumor size and growth when injected with Lewis lung carcinoma cells (LLCs) as compared to wild type mice.

Development

The S1pr2^f targeting vector was designed to insert a loxP site into the 5' UTR, upstream of the first coding exon (exon 2), of the sphingosine-1-phosphate receptor 2 (S1pr2) gene. A second loxP site was inserted downstream of the 3' UTR, followed by a cassette containing a frt-flanked PGK-neo and a third loxP site. The construct was electroporated into C57BL/6-derived embryonic stem (ES) cells and correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric mice were subsequently backcrossed to C57BL/6J mice for at least 7 generations. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. An aliquot of frozen sperm was used to fertilize oocytes from Sox2Cre mice (Stock No. 008454) to remove the neo cassette in rederived mice, leaving exon 2 floxed.

A 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. One of the 43 markers throughout the genome was segregating on Chromosome 13, likely a CBA segment leftover from the Edil3 insertion in the Sox2Cre mice.

Allele

Symbol: S1pr2^tm1.1Thla
Name: targeted mutation 1.1, Timothy Hla
MGI accession ID: MGI:6468214
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Synonyms: S1pr2^-
Marker symbol: S1pr2
Marker name: sphingosine-1-phosphate receptor 2
Marker synonyms: 1100001A16Rik; AGR16; DFNB68; EDG5; EDG-5; Gpcr13; GPCR18; H218; LPB2; S1P2; snGPCR18
Chromosome: 9
Strain of origin: C57BL/6
Molecular note: The targeting vector is designed to insert a loxP site into the 5' UTR, upstream of the first coding exon (exon 2). A second loxP site is inserted downstream of the 3' UTR, followed by a cassette containing an FRT-flanked PGK-neo and a third loxP site. Flp-mediated recombination removed the FRT-flanked neo cassette leaving exons 2 floxed.