B6;129-Eif2ak4^tm1.1Dron/J

Stock number: 008452
Research areas: Apoptosis Research, Cancer Research, Cell Biology Research, Immunology, Inflammation and Autoimmunity Research, Neurobiology Research, Research Tools

Description

GCN2.KO4^conditional (also called GCN2.KO4c) mice have loxP sites flanking exon XII of the eukaryotic translation initiation factor 2 alpha kinase 4 gene (Eif2ak4 or GCN2), and may be useful in generating conditional mutations for studying eIF2 (eukaryotic initiation factor 2) phosphorylation in response to environmental stresses (amino acid starvation, proteasome inhibition and UV irradiation).

Detailed description

Mice homozygous for the GCN2.KO4^conditional allele (also called GCN2.KO4c) are viable and fertile, with loxP sites flanking exon XII of the targeted gene. When bred to mice that express Cre recombinase, the resulting offspring will have this region (containing amino acid residues 606-648 encoding the critical lysine 618 required for kinase activity) deleted in the cre-expressing tissue(s). GCN2 is a protein kinase that phosphorylates eIF2 (eukaryotic initiation factor 2) in response to environmental stresses (amino acid starvation, proteasome inhibition and UV irradiation) resulting in a reduction of global translation and activation of stress-related transcription factors (such as NF-kappaB). These GCN2.KO4^conditional mice may be useful in studies related to eIF2 phosphorylation in response to environmental stresses.

When GCN2.KO4^conditional are bred to a strain expressing Cre recombinase in the central and peripheral nervous system (see Stock No. 003771 for example), this mutant mouse strain may be useful in studies of amino acid homeostasis.

Of note, mice with a traditional "knockout" (deletion of exon XII) are also available (see Stock No. 008240).

Development

A targeting vector was designed to place a loxP site just upstream of, and a loxP-flanked Neo resistance cassette just downstream of exon XII of the targeted gene. The construct was electroporated into 129S6/SvEvTac-derived W4 embryonic stem (ES) cells. Correctly targeted ES cells were then transiently transfected with a Cre recombinase expression plasmid. The resulting ES cells with the selection cassette removed and a single loxP site remaining on each side of exon XII were injected into recipient blastocysts to generate chimeric mice. The donating investigator reports that the resulting GCN2.KO4^conditional (also called GCN2.KO4c) mutant mice were then maintained on a mixed C57BL/6;129 genetic background and bred as homozygotes prior to arrival at The Jackson Laboratory. Upon arrival, mice were bred with C57BL/6J for at least one generation to establish the colony.

Allele

Symbol: Eif2ak4^tm1.1Dron
Name: targeted mutation 1.1, David Ron
MGI accession ID: MGI:3774318
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Eif2ak4
Marker name: eukaryotic translation initiation factor 2 alpha kinase 4
Chromosome: 2
Strain of origin: 129S6/SvEvTac
Molecular note: Exon 12 was flanked by loxP sites by inserting a loxP sequence into the EcorRI site in intron 11, and a floxed neomycin resistance cassette into the NheI site of intron 12. After homologous recombination, Cre was transiently expressed and ES cells were selected that had the neomycin cassette removed. Geneotypes were confirmed by PCR. The floxed exon 12 encodes a region of the kinase domain required for ATP binding. In addition, removal of this exon leads to a frameshift mutation with multiple stop codons.