Floxed cDGKζ KO mice possess loxP sites flanking exons 7-11 of the diacylglycerol kinase zeta (Dgkz) gene. This strain may be useful for studying diacylglycerol (DAG) regulation of TCR signaling and T cell activation.
Gary Koretzky, University of Pennsylvania
Floxed cDGKζ KO mice possess loxP sites flanking exons 7-11 of the diacylglycerol kinase zeta (Dgkz) gene. DGK is an enzyme that converts diacylglycerol (DAG) to phosphatidic acid, and can inhibit T signaling. DAG is essential for both T cell development and function. 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 exons 7-11 deleted in cre-expressing tissues. Cre recombination results in mice expressing a truncated 187 amino acid protein, which contains one of the DAG-binding C1 domains and a portion of the second C1 domain.
For example, cDGKζ KO floxed mice carrying a cre-inducible enhanced yellow fluorescent protein (EYFP) (B6.129X1-Gt(ROSA)26Sortm1(EYFP)Cos/J Stock No. 006148) were intercrossed to mice with ubiquitous expression of tamoxifen-inducible Cre recombinase (B6.Cg-Tg(UBC-cre/ERT2)1Ejb/2J Stock No. 008085). The resulting DGKζ KO mice lack DGKζ in T cells. T regulatory cells (YFP+CD25+CD4+) from DGKζ KO mice exhibit an enhanced ability to limit
conventional CD4+ T cell proliferation in vitro.
A targeting vector was designed to insert a loxP-flanked neomycin resistance (neo) cassette upstream of exon 7, and a single loxP site downstream of exon 11 of the diacylglycerol kinase zeta (Dgkz) gene. The construct was electroporated into C57BL/6 embryonic stem (ES) cells. Correctly targeted ES cells were injected into Balb/c blastocysts and resulting chimeric males were bred with C57BL/6 females. Resulting offspring were bred to establish a colony of cDGKζ KO mice. 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).
|Allele Name||targeted mutation 2, Gary A Koretzky|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Gene Symbol and Name||Dgkz, diacylglycerol kinase zeta|
|Strain of Origin||C57BL/6|
|Molecular Note||The targeting vector is designed to insert a loxP-flanked neomycin resistance (neo) cassette upstream of exon 7, and a single loxP site downstream of exon 11.|
When maintaining a live colony mice homozygous for the floxed allele may be bred together.
When using the cDGKζ KO mouse strain in a publication, please cite the originating article(s) and include JAX stock #029130 in your Materials and Methods section.