B6.129S4-Cxcr3^tm1Arsa/SoghJ

Stock number: 023337
Product name: CIBER
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

These knock-in mice express EGFP from the endogenous X-linked Cxcr3 gene. They may be useful for tracking Cxcr3-expressing immune cells during infection and disease progression.

Detailed description

In this CIBER strain, an internal ribosome entry site (IRES), an enhanced green fluorescent protein (EGFP), and bovine growth hormone polyadenylation sequence were placed downstream of exon 2 of the X-linked chemokine (C-X-C motif) receptor 3 (Cxcr3) gene. Homozygous females and hemizygous males are viable and fertile. Cxcr3 encodes a chemokine receptor involved in immunity, autoimmunity, and progression of diseases such as cancer, and atherosclerosis. In these CIBER mice, GFP fluorescence is seen on activated Th1 cells, CD8⁺ T cells, NK cells, NKT cells, natural regulatory T cells, as well as innate CD8⁺ T cells.

Development

A targeting vector was designed to insert an internal ribosome entry site (IRES), an enhanced green fluorescent protein (EGFP), and bovine growth hormone polyadenylation sequence, followed by a frt-flanked neomycin resistance cassette, downstream of the X-linked chemokine (C-X-C motif) receptor 3 (Cxcr3) gene. The construct was electroporated into 129S4/SvJae-derived J1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and the resulting chimeric mice were bred to C57BL/6-Tg(Zp3-cre)93Knw/J transgenic mice (Stock No. 003651) to delete the neo cassette. Offspring were crossed to remove the Flp-expressing transgene. These mice were bred to C57BL/6 mice for at least 10 generations (see SNP note below). Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J (Stock No. 000664) for at least one generation to establish the colony.

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 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 27 markers throughout the genome suggested a C57BL/6 genetic background, all 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 mixed C57BL/6J ; C57BL/6N genetic background.

Allele

Symbol: Cxcr3^tm1Arsa
Name: targeted mutation 1, Abhay R Satoskar
MGI accession ID: MGI:5478545
Generation method: Targeted
Attributes: Reporter
Marker symbol: Cxcr3
Marker name: chemokine (C-X-C motif) receptor 3
Chromosome: X
Strain of origin: 129S4/SvJae
Expressed genes: GFP,Green Fluorescent Protein,
Site of expression: GFP fluorescence is seen on activated Th1 cells, CD8+ T cells, NK cells, NKT cells, natural regulatory T cells, as well as innate CD8+ T cells.
Molecular note: An IRES, EGFP and floxed pgk-neo cassette were inserted in the 3' untranslated region of exon 2. Protein expression in activated T cells from mutant mice is similar to wild-type controls. Expression pattern of EGFP matches endogenous protein expression.