STOCK Prkaa2^tm1.1Sjm/J

Stock number: 014142
Product name: Prkaa2^fl
Research areas: Cancer Research, Cell Biology Research, Developmental Biology Research, Research Tools

Description

These Prkaa2 mutant mice possess loxP sites flanking exon 2 of the protein kinase, AMP-activated, alpha 2 catalytic subunit (Prkaa2) gene. This strain may be useful for studying cell growth and energy expenditure, as well as bladder cancer.

Detailed description

These Prkaa2 mutant mice possess loxP sites flanking exon 2 of the protein kinase, AMP-activated, alpha 2 catalytic subunit (Prkaa2) gene. Mice that are homozygous for this allele are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. Prkaa2 belongs to the serine/threonine protein kinase family and is the catalytic subunit of the 5'-prime-AMP-activated protein kinase (AMPK). AMPK is a sensor of the energy status of cells and ensures survival during stress. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exon 2 deleted in the cre-expressing tissues. When bred to transgenic mice expressing Cre recombinase driven by myxovirus (influenza virus) resistance 1 (Mx1) promoter/enhanced elements, PRKAA2 expression is abolished in hematopoietic cells. This strain may be useful for studying cell growth and energy expenditure, as well as bladder cancer.

Development

A targeting vector was designed to insert a loxP site upstream of exon 2, and a Frt-flanked neomycin resistance cassette (neo) followed by a second loxP site downstream of exon 2 of the protein kinase, AMP-activated, alpha 2 catalytic subunit (Prkaa2) gene. The construct was electroporated into Bruce4-derived C57BL/6 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts from B6(Cg)-Tyr^c-2J/J mice (Stock No. 000058) and the resulting chimeric males were bred to female B6(Cg)-Tyr^c-2J/J mice to establish a colony. These mice were then bred to mice carrying Tg(ACTFLPe)9205Dym to delete the neo cassette. Offspring were then backcrossed to C57BL/6 for at least four generations (see SNP results below) prior to sending males to The Jackson Laboratory Repository. Upon arrival, males were used to cryopreserve sperm. To establish the living mouse colony, an aliquot of the frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the living colony at The Jackson Laboratory Repository. This revealed 3 of 27 markers, one each on chromosomes 9, 11, and 13, that were not fixed for C57BL/6 allele-type (e.g.: still segregating for other genetic background). The source of this genetic contribution is not known. In addition, all 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating (on chromosomes 8, 11, 13, 15 and 19). These data suggest the mice may have been backcrossed to C57BL/6 for fewer generations than reported prior to arrival at The Jackson Laboratory.

Allele

Symbol: Prkaa2^tm1.1Sjm
Name: targeted mutation 1.1, Sean J Morrison
MGI accession ID: MGI:4836201
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Prkaa2
Marker name: protein kinase, AMP-activated, alpha 2 catalytic subunit
Chromosome: 4
Strain of origin: B6.Cg-Thy1^a
Molecular note: A loxP site was inserted upstream of exon 2 and an FRT-flanked neo cassette with a 3' loxP site was inserted downstream of exon 2. Flp-mediated recombination removed the neo selection cassette.