B6.Cg-Prkch^tm1.2Gasc/J

Stock number: 018988
Research areas: Immunology, Inflammation and Autoimmunity Research

Description

PKCeta^-/- mice may be useful for studying T cell homeostasis and activation.

Detailed description

Exon 2 of the protein kinase C, eta (Prkch) gene was removed, abolishing gene function. These PKCeta^-/- homozygotes are viable and fertile. Prkch-encoded PKCeta is a serine/threonine kinase abundant in T cells which is recruited during positive selection of thymocytes. T-cells from these PKCeta^-/- mice showed poor proliferation in response to antigen stimulation and exhibited defective homeostatic proliferation. These mice also have a higher ratio of CD4⁺ to CD8⁺ T cells compared to that of wild-type mice.

Development

A targeting vector was designed to insert a loxP site, followed by a frt-flanked neomycin resistance (neo) cassette, upstream of exon 2 and a second loxP site downstream of exon 2 of the protein kinase C, eta (Prkch) gene. The construct was electroporated into C57BL/6-derived Brunce-4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric males were bred to C57BL/6J females. Offspring were bred with B6.Cg-Tg(ACTFLPe)9205Dym/J (Stock No. 005703) to delete the neo cassette, then the progeny were bred with B6.FVB-Tg(EIIa-cre)C5379Lmgd/J (Stock No. 003724) transgenic mice to delete exon 2. progeny were crossed to remove the cre- and Flp-expressing transgenes. These mice were then backcrossed to C57BL/6J mice for at least 4 generations. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.

Allele

Symbol: Prkch^tm1.2Gasc
Name: targeted mutation 1.2, Nicholas Gascoigne
MGI accession ID: MGI:5431008
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Prkch
Marker name: protein kinase C, eta
Chromosome: 12
Strain of origin: B6.Cg-Thy1^a
Molecular note: A targeting vector was designed to insert a loxP site, followed by a frt-flanked neomycin resistance (neo) cassette, upstream of exon 2 and a second loxP site downstream of exon 2. Flp-mediated recombination removed the neo cassette while cre-mediated recombination remvoed exon 2.