B6.129S1-C5ar2^tm1Cge/HgrJ

Stock number: 033608
Product name: C5L2^-
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

C5L2- mice have a green fluorescent protein sequence and a neomycin resistance (neo) cassette replacing 1 kb of the coding sequence of the complement component 5a receptor 2 (C5ar2) gene abolishing gene expression. These mice may be useful when studying T_REG generation associated with the onset of inflammatory diseases.

Detailed description

C5L2- mice have a green fluorescent protein sequence and a neomycin resistance (neo) cassette replacing 1 kb of the coding sequence of the complement component 5a receptor 2 (C5ar2) gene abolishing gene expression. C5L2 is a G-protein coupled receptor 1 family member involved in the complement system of the immune response. Expressed on immune cells, C5aR2 modulates various inflammatory diseases in mice, including sepsis, asthma, cystic fibrosis, and chronic obstructive lung disease, and has been shown to facilitate regulatory T cell (T_REG) generation in vitro and in vivo. Homozygous mice are viable and fertile. Little to no EGFP fluorescence is detected in unstimulated T cells from these mice. After a 5 day stimulation of splenic T cells with anti-CD3 and TGF-β/IL-2, EGFP fluorescence is detectable by flow cytometry. Adoptive transfer of naive CD4⁺ T cells from these mice into Rag1^-/- recipients results in fewer _REG cells compared to wild type controls.

Development

The C5L2- targeting vector was designed to replace 1 kb of coding sequence of the complement component 5a receptor 2 (C5ar2) gene with a green fluorescent protein sequence and a neomycin resistance (neo) cassette. The construct was electroporated into 129S1/Sv-Oca2⁺ Tyr⁺ Kitl⁺-derived CJ7 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and the resulting chimeric males were bred to C57BL/6 females. Mice were subsequently backcrossed to C57BL/6 mice for at least 5 generations by the donating lab (see SNP note below). Upon arrival at The Jackson Laboratory Repository, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).



In 2019 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. While the 43 markers throughout the genome suggested a C57BL/6 genetic background, 1 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.

Allele

Symbol: C5ar2^tm1Cge
Name: targeted mutation 1, Craig Gerard
MGI accession ID: MGI:3612426
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: C5L2^-
Marker symbol: C5ar2
Marker name: complement component 5a receptor 2
Marker synonyms: C5L2; E030029A11Rik; GPF77; Gpr77
Chromosome: 7
Strain of origin: 129S1/Sv-Oca2⁺ Tyr⁺ Kitl⁺
Molecular note: Approximately 1 kb of coding sequence was replaced with genes encoding green fluorescent protein and neomycin resistnace under the control of the pgk promoter. Transcript was not detected by RT-PCR of mutant bone marrow.