B6(SJL)-Il10ra^tm1.1Tlg/J

Stock number: 028146
Product name: IL-10Rα^flox
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

These floxed mutant mice possess loxP sites flanking exon 3 of the Il10ra gene. This strain may be useful for generating conditional mutations in applications related to the study of the role played by the IL10 pathway in autoimmunity such as severe inflammatory bowel syndrome.

Detailed description

The interleukin 10 receptor alpha subunit encoded by the Il10ra gene binds IL10 with high affinity, and to the tyrosine kinase JAK1, mediating immune response through IL10 signaling. These IL-10Rα^flox mice possess loxP sites on either side of exon 3 of the targeted gene. Mice that are homozygous for this allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exon 3 deleted in the cre-expressing tissues. Exon 3 encodes most of the IL-10 interacting (cytokine-binding) domain. Removal of the floxed sequence creates a null allele.

When bred to a strain with Cre recombinase expression in developing T cells (see Stock No. 022071 for example), this mutant mouse strain may be useful in studies of the autoimmune response.

 

Development

A targeting vector containing a FRT site-flanked NEO cassette was used to introduce loxP sites on either side of exon 3. The FRT site-flanked cassette was inserted into intron 3 (upstream of exon 3). The construct was electroporated into C57BL/6 derived ES26.2 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6J-Tyr^c-2J blastocysts. The resulting chimeric animals were tested for germline transmission. The mice were crossed to B6.Cg-Tg(ACTFLPe)9205Dym/J mice (Stock No. 005703) to remove the NEO cassette. The mice were then backcrossed to C57BL/6 for 8 to 10 generations. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

In 2020, 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. Two of the 5 markers that determine C57BL/6J from C57BL/6N, including the rd8 recessive mutation associated with retinal degeneration on Chromosome 1 within the Crb1 locus, were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.

Allele

Symbol: Il10ra^tm1.1Tlg
Name: targeted mutation 1.1, Terrence L Geiger
MGI accession ID: MGI:5450726
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Il10ra
Marker name: interleukin 10 receptor, alpha
Chromosome: 9
Strain of origin: C57BL/6
Molecular note: An FRT flanked PGK neo resistance cassette was inserted in intron 2 and loxP sites were inserted flanking exon 3. Subsequent expression of flp recombinase removed the selection cassette leaving exon 3 flanked by loxP sites. Exon 3 contains much of the Il10 interacting domain. Genomic loss of exon 3 was observed in T cells expressing cre under the control of the Cd4 promoter by PCR analysis.