STOCK Eif2ak3^tm1.2Drc/J

Stock number: 023066
Product name: PERK^loxP
Research areas: Cell Biology Research, Developmental Biology Research, Diabetes and Obesity Research, Internal/Organ Research, Research Tools

Description

These floxed mutant mice possess loxP sites flanking exon 7-9 of the Eif2ak3 gene. They may be useful in applications related to the study of protein synthesis regulation in secretory organs, as well as Wolcott-Rallison syndrome and Alzheimer's disease.

Detailed description

These Perk^loxP/loxP mutant mice possess loxP sites flanking exons 7-9 of the eukaryotic translation initiation factor 2 alpha kinase 3 (Eif2ak3) gene. PERK is an endoplasmic reticulum (ER) transmembrane protein, expressed mainly in secretory and endocrine organs such as the pancreas. Mutations in PERK have been implicated in the onset of Wolcott-Rallison syndrome which is characterized by early onset of insulin-dependent diabetes, dwarfism, and skeletal dysplasias. Mice that are homozygous for this allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting double mutant offspring will have exons 7-9 deleted in cre-expressing tissues.

For example, when crossed to B6.FVB-Tg(EIIa-cre)C5379Lmgd/J mice (Stock No. 003724), resulting Perk^-/- mice exhibit permanent neonatal diabetes, exocrine pancreas atrophy, growth retardation, severe osteopenia, skeletal dysplasia, recurrent hepatitis, and behavior/neurological dysfunctions.

 

Development

A targeting vector was designed to insert a single loxP site upstream of exon 7, and a loxP-flanked neomycin resistance (neo) cassette downstream of exon 9 of the eukaryotic translation initiation factor 2 alpha kinase 3 (Eif2ak3) gene. The construct was electroporated into 129S6/SvEvTac-derived TL1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6J blastocysts and resulting chimeric mice were bred with B6.FVB-Tg(EIIa-cre)C5379Lmgd/J mice transgenic mice (Stock No. 003724) to delete the neo cassette. Resulting mice contained multiple gene rearrangements; intact floxed-exons 7-9, intact floxed-neo cassette, or excision of both exons 7-9 and the neo cassette. Resulting mice, containing only the floxed-exons, were crossed to remove the cre-expressing transgene. These Perk^loxP/loxP mice were bred to C57BL/6J mice for at least ten generations (see SNP note below). Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation.

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 rederived living colony at The Jackson Laboratory Repository. Three markers throughout the genome were still segregating for 129 suggesting an incomplete backcross.

 

Allele

Symbol: Eif2ak3^tm1.2Drc
Name: targeted mutation 1.2, Douglas R Cavener
MGI accession ID: MGI:5502572
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Eif2ak3
Marker name: eukaryotic translation initiation factor 2 alpha kinase 3
Chromosome: 6
Strain of origin: 129S6/SvEvTac
Molecular note: Two loxP sites were introduced to intronic regions flanking the three exons encoding part of the lumenal domain, the transmembrane domain, and part of the catalytic domain. A third loxP site was introduced with a PGK-neo cassette directly downstream of the second loxP site in the same intron. Cre-mediated recombination removed the neo cassette and left the three exons floxed.