B6.129S1-Ripk2^tm1Flv/J

Stock number: 007017
Product name: RIP2 KO
Research areas: Immunology, Inflammation and Autoimmunity Research

Description

Production of macrophage cytokines in response to stimulation with lipopolysaccharide, peptidoglycan and double-stranded RNA is reduced in these Ripk2 KO mice, making them suitable for use in studies of innate and adaptive immune system signaling.

Detailed description

Rip2 KO mice contain a neo cassette in reverse orientation in place of exons 2-3 of the Ripk2 (receptor (TNFRSF)-interacting serine-threonine kinase 2) gene. Cytokine production in macrophages is reduced in homozygotes upon stimulation with lipopolysaccharide, peptidoglycan and double-stranded RNA, but not with bacterial DNA. RIPK2 deficient cells are also hyporesponsive to signaling through IL1 and IL18 interleukin receptors, and deficient for signaling through NOD (nucleotide-binding oligomerization domain) proteins. T cells show severely reduced NFKB (nuclear factor of kappa light chain gene enhancer in B-cells) activation, IL2 production and proliferation on T cell receptor (TCR) engagement, and impaired differentiation to T-helper subtype 1 (T_H1)cells. Western blot analysis of thymocytes demonstrates the absence of protein in homozygous mutant mice. Homozygotes are viable and fertile.

Development

A targeting vector was designed to replace exons 2-3 of the receptor (TNFRSF)-interacting serine-threonine kinase 2 (Ripk2) gene, encoding the active site aspartate residue, with a loxP-flanked neomycin resistance cassette in the opposite orientation to the gene. The vector was linearized and injected into 129S1/Sv-derived W9.5 embryonic stem (ES) cells. Targeted clones were injected into C57BL/6 blastocysts. Resultant chimeric mice were backcrossed to C57BL/6 for 10 generations by the donating laboratory (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 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. One of the 43 markers on Chromosome 4 is segregating, likely due to targeted mutation. 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 C57BL/6N genetic background.

 

Allele

Symbol: Ripk2^tm1Flv
Name: targeted mutation 1, Richard A Flavell
MGI accession ID: MGI:2660793
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Ripk2
Marker name: receptor (TNFRSF)-interacting serine-threonine kinase 2
Chromosome: 4
Strain of origin: 129S1/Sv-Oca2⁺ Tyr⁺ Kitl⁺
Molecular note: The gene was disrupted by replacement of exons 2 and 3 with a neomycin resistance gene by homologous recombination. Absence of gene expression in homozygous mutant animals was confirmed by Western blot analysis of thymocyte extracts.