Prkcg^em1Jpka

CRISPR/Cas9 technology generated a C to A change at position 27 (c.71C>A) resulting in an alanine to glutamate substitution at amino acid 24 (p.A24E). In addition, a G to A change at position 78 (c.78G>A) that does not change the amino acid (p.R26R) was introduced to prevent the donor DNA from being a suitable target for cas9 cleavage. The A24E mutation keeps PKC gamma in a constitutive active open conformation. mRNA expression is comparable to wild-type but protein is strongly reduced in homozygotes. Cycloheximide trace assay shows increased protein degradation in these mice. Heterozygotes and homozygotes show increased PKC kinase activity despite decreased protein levels. (J:305637)