STOCK Ubr4^tm1.2Nkt/MdnJ

Stock number: 024844
Product name: p600 flox
Research areas: Developmental Biology Research, Neurobiology Research, Research Tools

Description

These p600^flox/flox mice may be useful for studying the embryonic expression of p600.

Detailed description

These p600^flox/flox mutant mice possess loxP sites flanking exon 1 of the ubiquitin protein ligase E3 component n-recognin 4 (Ubr4) gene. P600 is involved in cytoskeletal organization, membrane morphogenesis, integrin-mediated survival signaling, calcium-calmodulin signaling, and the N-end rule pathway of ubiquitin-proteasome-mediated proteolysis. Embryonic p600 is essential for liver, cardiac, and brain development. Mice that are homozygous for this allele are viable and fertile. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exon 1 deleted in the cre-expressing tissues.

For example, when bred to B6.Cg-Tg(Sox2-cre)1Amc/J mice (Stock No. 008454) expressing Cre-recombinase under the Sox2 promoter, resulting mice die by E14.5 due to severe cardiac problems including ventricular septal defects and thin ventricular walls. Brain of the knockout mice displays thinner cortical plate with significant less Tuj1-positive neurons. Starting from E13.5, the animals display enlarged ventricles. The orientation of the mitotic spindle in cortical neural progenitors is randomized in E12.5 animals.

Development

A targeting vector was designed to insert a loxP site followed by a frt-flanked neomycin resistance (neo) cassette upstream of exon 1, and a second loxP site downstream of exon 1 of the ubiquitin protein ligase E3 component n-recognin 4 (Ubr4) gene. The construct was electroporated into 129S6/SvEvTac-derived TC1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric males were bred with C57BL/6NCrl females. Offspring were bred with B6;SJL-Tg(ACTFLPe)9205Dym/J transgenic mice (Stock No. 003800) to delete the neo cassette, and progeny were crossed to remove the Flp-expressing transgene. The donating investigator reported that these p600^flox/flox mice were bred to C57BL/6NCrl mice for at least 15 generations (See SNP note below). Upon arrival, mice were bred to C57BL/6NJ inbred mice (Stock No. 005304) for at least one generation to establish the colony.

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. 5 markers on three choromosomes were segregating, suggesting an incomplete backcross.

Allele

Symbol: Ubr4^tm1.2Nkt
Name: targeted mutation 1.2, Yoshihiro Nakatani
MGI accession ID: MGI:5532332
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Ubr4
Marker name: ubiquitin protein ligase E3 component n-recognin 4
Chromosome: 4
Strain of origin: 129S6/SvEvTac
Molecular note: An FRT-flanked neomycin resistance cassette with a 5' loxP site was inserted upstream of exon 1. An additional loxP site was inserted downstream of exon 1. Flp-mediated recombination removed the neomycin resistance cassette and left exon 1 floxed.