129S1/SvImJ-Nlrc4^em4Vnce/J

Stock number: 039846
Product name: Nlrc4^152stop, Nlrc4^-
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

Although the infectious dose for Shigella in humans can be as low as 10-100 bacteria, wildtype mice are resistant to high doses of oral Shigella challenge. Nlrc4^152stop (Nlrc4^-) mice are susceptible to oral Shigella challenge and provide a physiologically relevant model of human shigellosis.

Note that 129-derived mice also harbor a null mutation in the Casp4 (caspase 4, apoptosis-related cysteine peptidase; also called Casp11) gene, so this strain is effectively double deficient for both Nlrc4 and Casp4. As a result, the mice exhibit heightened susceptibility to Shigella infection as compared to Casp4-sufficient Nlrc4-deficient mice on the C57BL/6J background (see Stock No. 039847).

Detailed description

The NAIP-NLRC4 inflammasome is a cytosolic immune sensor expressed in immune cells and intestinal epithelial cells of mice. Its function is to recognize bacterial ligands (such as flagellin, rod, and needle) and initiate downstream immune responses, such as cytokine release and pyroptosis.

Nlrc4^152stop (Nlrc4^-) mice carry a targeted knock-out of the mouse NLR family, CARD domain containing 4 (Nlrc4) gene, a critical component of the NAIP-NLRC4 inflammasome. A CRISPR/cas9-generated 10 bp deletion in exon 5 of the mouse Nlrc4 gene created a frame shift mutation and a premature stop codon at amino acid position 152 that results in a loss of protein function.

Mice lacking NLRC4 breed normally and appear healthy in specific pathogen-free conditions. However, they are susceptible to systemic and gastrointestinal bacterial infection. Nlrc4^152stop mice are susceptible to oral Shigella challenge and provide a physiologically relevant model of human shigellosis (Mitchell, 2020; PMID: 33074100).

Note that 129-derived mice also harbor a null mutation in the Casp4 (caspase 4, apoptosis-related cysteine peptidase; also called Casp11) gene, so this strain is effectively double deficient for both Nlrc4 and Casp4. As a result, the mice exhibit heightened susceptibility to Shigella infection as compared to Casp4-sufficient Nlrc4-deficient mice on the C57BL/6J background (see Stock No. 039847).

Development

A CRISPR/cas9-generated 10 bp deletion in exon 5 of the mouse Nlrc4 gene (gRNAs: 5'-CAGGTCACAGAAG AAGACCTGAATG3' and 5'CACCTGGACTCCTGGATTTGG-3') created a frame shift mutation and a premature stop codon at amino acid position 152 that results in a loss of protein function. The mutation was created in 129S1/SvImJ (Stock No. 002448) embryos which are naturally deficient in Casp4 (also called Casp11) expression. Founders carrying mutations were bred one generation to wildtype mice to separate modified haplotypes. Homozygous lines were generated by interbreeding heterozygotes carrying matched haplotypes. Upon arrival, sperm was cryopreserved. To establish a live colony, an aliquot of frozen sperm was used to fertilize 129S1/SvImJ oocytes (Stock No. 002448).

Allele

Symbol: Nlrc4^em4Vnce
Name: endonuclease-mediated mutation 4, Russell E Vance
MGI accession ID: MGI:7709165
Generation method: Endonuclease-mediated
Attributes: Null/Knockout
Synonyms: Nlrc4^-
Marker symbol: Nlrc4
Marker name: NLR family, CARD domain containing 4
Marker synonyms: 9530011P19Rik; AIFEC; Card12; CLAN; CLAN1; CLANA; CLANB; CLANC; CLAND; CLR2.1; FCAS4; IPAF
Chromosome: 17
Strain of origin: 129S1/SvImJ
Molecular note: A CRISPR/cas9-generated 10 bp deletion in exon 5 of the mouse Nlrc4 gene created a frame shift mutation and a premature stop codon at amino acid position 152 that results in a loss of protein function. The mutation was created in 129S1/SvImJ embryos.