NOD.129S1(B6)-Irak3^tm1Flv/LwnJ

Stock number: 022836
Product name: IRAK-M-/-NOD
Research areas: Diabetes and Obesity Research, Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

These Irak3 knock out mice on the NOD background develop spontaneous Type I diabetes mellitus with an onset as early as 6 of 7 weeks of age.

Detailed description

Mice that are homozygous for the targeted mutation are viable and fertile. On the NOD background, female homozygotes develop spontaneous Type I diabetes mellitus with an onset as early as 6 of 7 weeks of age. Male homozygotes on the NOD background also show a tendency for earlier onset of diabetes. Circulating anti-insulin antibody levels are elevated, more pancreatic islets are infiltrated, there is a higher proportion of effector/memory cells, and an increase in CD4+ T cells in pancreatic draining lymph nodes, in female homozygotes, when compared to controls.

Development

The targeting vector was designed to replace a 1.2 kb region of the gene (containing three exons encoding two-thirds of the kinase domain) with a loxP-flanked neomycin resistance cassette. The targeting vector was linearized and electroporated into 129S1/Sv-derived W9.5 embryonic stem (ES) cells. Targeted clones were injected into C57BL/6 blastocysts and resulting chimeric males were bred to C57BL/6 females. Interbreeding of heterozygotes was performed to generate homozygotes. This line was backcrossed to C57BL/6 for approximately 6 generations by the originating laboratory. The mice were obtained by Dr. Li Wen and backcrossed to NOD/Caj (Yale University) for 11 generations, and then intercrossed for 8 generations. Upon arrival at The Jackson Laboratory, the mice were crossed to NOD/ShiLtJ (Stock No. 001976) at least once to establish the colony.

Allele

Symbol: Irak3^tm1Flv
Name: targeted mutation 1, Richard A Flavell
MGI accession ID: MGI:2384197
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Irak3
Marker name: interleukin-1 receptor-associated kinase 3
Chromosome: 10
Strain of origin: 129S1/Sv-Oca2⁺ Tyr⁺ Kitl⁺
Molecular note: 1.2 kb of sequence were replaced with a floxed neo cassette. The deleted region contained 3 exons encoding two thirds of the kinase domain. While Western blot analysis of LPS-stimulated bone marrow macrophage extracts using a carboxy terminal antibody confirmed the absence of normal protein in homozygous mutant mice, truncated transcript was detected by Northern blot analysis using an amino terminal probe. Sequence analysis of RT-PCR products revealed an in frame stop codon generated by aberrant splicing of the endogenous gene and the neo transgene.