C57BL/6-Prkcd^tm1.1Chou/J

Stock number: 028437
Product name: AS PKCδ
Research areas: Cell Biology Research

Description

AS PKCδ mice contain a mutation in the Prkcd gene, allowing PKCδ to be regulated by specific kinase inhibitors. These mice are useful when studying PKC&delta substrates for mapping downstream signaling pathways.

Detailed description

AS PKCδ mice contain the amino acid mutation M425A in exon 13 of the endogenous protein kinase C, delta ( Prkcd) gene. PKCδ plays a role in stroke-reperfusion injury by stimulating the infiltration of neutrophils to the ischemic brain tissues. Neutrophils release free radicals, proteases, and cytokines to promote further recruitment of neutrophils and other leukocytes to the site of injury. These inflammatory responses lead to secondary damage to the effected tissue. The M425A mutation introduces a conformational change in the “gatekeeper residue” which lies in the middle of the catalytic domain and is a key residue for determining the selectivity of kinase inhibitors. This AS mutation allows AS PKCδ to use ATP analogs, such as N6-(benzyl)-ATP, and can be specifically inhibited by and PP1 analogs, such as , such as N6-(benzyl)-ATP, 1NA-PP1 and 2MB-PP1.

Development

A targeting construct was designed to insert a loxP-flanked neomycin (neo) resistance cassette downstream of exon 13 of the protein kinase C, delta (Prkcd) gene. A point mutation was introduced in exon 13, M425A, which encodes a key residue for determining the selectivity of PKCδ inhibitors. This targeting construct was electroporated into C57BL/6-derived embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a Cre expression plasmid to delete the neo cassette and resulting ES cells were injected into blastocysts. The resulting chimeric mice were bred to C57BL/6NTac mice for at least 10 generations. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes (Stock No. 005304).

Allele

Symbol: Prkcd^tm1.1Chou
Name: targeted mutation 1.1, Wen-Hai Chou
MGI accession ID: MGI:5699182
Generation method: Targeted
Attributes: Not Applicable
Marker symbol: Prkcd
Marker name: protein kinase C, delta
Chromosome: 14
Strain of origin: C57BL/6
Molecular note: The targeting construct was designed to insert a loxP-flanked neomycin (neo) resistance cassette downstream of exon 13 of the gene. A point mutation, M425A, is introduced in exon 13, which encodes a key residue for determining the selectivity of PKC inhibitors. Cre-mediated recombination removed the floxed neo cassette.