C.129S1(B6)-C5ar2^tm1Cge/BaoluJ

Stock number: 033906
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 TREG 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.

Of note, this allele is also available on a congenic C57BL/6J background as Stock No. 033608.

In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. It should be noted that the phenotype of these BALB/c congenic mice could vary from that originally described on a C57BL/6J congenic background. We may modify the strain description if necessary as published results become available.

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 five generations before arriving at The Jackson Laboratory. These mice are available as Stock No. 033608. Some of these mice were further backcrossed to BALB/cJ mice (Stock No. 000651) for 12 generations by Dr. Craig Gerard (Boston Children's Hospital) before being sent back to The Jackson Laboratory. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize BALB/cJ oocytes.

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.