B6(Cg)-Pgam5^{tm1c(EUCOMM)Wtsi}/FkmcJ

Stock number: 032172
Product name: Pgam5^fl
Research areas: Immunology, Inflammation and Autoimmunity Research, Metabolism Research, Neurobiology Research, Research Tools

Description

Pgam5^fl mice have exon 2 of the phosphoglycerate mutase family member 5 (Pgam5) gene floxed, making these mice useful for studying inflammasome activation and function.

Detailed description

Pgam5^fl floxed mice have loxP sites flanking exon 2 of the phosphoglycerate mutase family member 5 (Pgam5) gene. PGAM5 is a mitochondrial phosphatase that has been reported to mediate some forms of necrosis. Moreover, PGAM5 is involved in LPS-induced inflammasome activation, IL-1Β secretion, and mitophagy. Mice that are homozygous for this allele are viable and fertile. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exon 2 deleted in the cre-expressing tissues.

When bred to Sox2-Cre mice, with germline cre recombinase expression (Stock No. 008454), resulting mice are smaller than wildtype littermates and exhibit reduced viability into adulthood. Bone marrow derived macrophages (BMDMs) from Pgam5^-/- mice display a reduction in mitochondrial volume and produce reduced level of ROS in response to inflammasome activation.

 

Development

The L1L2_Bact_P cassette was inserted at a position on Chromosome 5 upstream of exon 2 of the phosphoglycerate mutase family member 5 (Pgam5) gene. The cassette is composed of a frt site followed by a lacZ sequence and a loxP site. This first loxP site is followed by neomycin resistance cassette under the control of the human beta-actin promoter, SV40 polyA, a second frt site and a second loxP site. A third loxP site was inserted downstream of exon 2. The construct, clone EPD0226_5_H02, was introduced into C57BL/6N-derived JM8.N4 embryonic stem (ES) cells, and correctly targeted ES cells were injected into B6(Cg)-Tyr^c-2J/J blastocysts. The resulting chimeric males were bred to C57BL/6J mice (Stock No. 000664). Subsequently these mice were bred to ROSA26::FLPe mice on a C57BL6 background to remove the frt-flanked lacZ sequence and neomycin cassette. Resulting Pgam5^fl offspring were bred to C57BL/6J mice for at least 5 generations. Cryopreserved embryos were shipped to The Jackson Laboratory. An aliquot of frozen embryos were used to establish our live colony.

Allele

Symbol: Pgam5^{tm1c(EUCOMM)Wtsi}
Name: targeted mutation 1c, Wellcome Trust Sanger Institute
MGI accession ID: MGI:5882559
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Synonyms: Pgam5^fl; Pgam5^flox
Marker symbol: Pgam5
Marker name: phosphoglycerate mutase family member 5
Marker synonyms: 2610528A17Rik; BXLBV68; RGD1312028
Chromosome: 5
Strain of origin: C57BL/6N
Molecular note: The L1L2_Bact_P cassette was inserted at position 110414719 of Chromosome 5 upstream of the critical exon (exon 2) (Build GRCm39). The cassette is composed of an FRT site followed by lacZ sequence and a loxP site. This first loxP site is followed by neomycin resistance gene under the control of the human beta-actin promoter, SV40 polyA, a second FRT site and a second loxP site. A third loxP site is inserted downstream of the targeted exon 2 at position 110415572. Exon 2 is thus flanked by loxP sites. A "conditional ready" (floxed) allele was created by flp recombinase expression in mice carrying this allele to remove the lacZ sequence and neo selection cassette, leaving exon 2 flanked by loxP sites.