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.
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 Name||targeted mutation 1.2, Nicholas Gascoigne|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Prkch, protein kinase C, eta|
|Strain of Origin||B6.Cg-Thy1a|
|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.|
When maintaining a live colony, homozygous mice may be bred together.
When using the B6.Cg-Prkchtm1.2Gasc/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #018988 in your Materials and Methods section.