B6;129S-Ate1^tm2.1Akas/J

Stock number: 032519
Product name: Ate1 floxed
Research areas: Cell Biology Research, Developmental Biology Research, Research Tools

Description

Ate1-floxed mice possess loxP sites flanking exons 1-3 of the arginyltransferase 1 (Ate1) gene. These floxed mice are useful when studying the role of ATE1 in gametogenesis and early embryonic development.

Detailed description

Ate1-floxed mice possess loxP sites flanking exons 1-3 of the arginyltransferase 1 (Ate1) gene. Ate1 encodes an enzyme that is involved in posttranslational conjugation of arginine to proteins, a modification implicated in ubiquitin-dependent degradation, autophagy, stress response, and protein-protein interactions. It is essential for cardiovascular development, actin cytoskeleton functioning, tissue morphogenesis, and cell migration. 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 exons 1-3 deleted in the cre-expressing tissues.

When bred to Tek-cre mice, expressing Cre Recombinase in endothelial cells (ECs), resulting Tek-Ate1 knockout mice are viable and have no visible angiogenesis-related phenotypes. However, these mice exhibit reproductive defects, with high rates of embryonic lethality in the second generation, up to 80%, due to defects in gametogenesis.

 

Development

The Ate1-floxed targeting vector was designed to insert a loxP site upstream of exon 1, and a frt-flanked neomycin selection cassette, followed by a second loxP site, downstream of exon 3 of the arginyltransferase 1 (Ate1) gene. The construct was electroporated into (129S6/SvEvTac x C57BL/6J)F1-derived D1 embryonic stem (ES) cells. Correctly targeted ES cells were aggregated with recipient 8-cell CD-1 embryos and chimeric males were bred to ROSA26::FLPe females (Stock No. 003946) to remove the neo cassette. Progeny were crossed to remove the Flpe-expressing transgene, and offspring were bred to C57BL/6 mice for at least 4 generations (see SNP note below). Upon arrival at The Jackson Laboratory Repository, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).



A 48 SNP (single nucleotide polymorphism) panel analysis, with 43 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. 12 of the 43 markers throughout the genome were segregating with 129, suggesting an incomplete backcross.

Allele

Symbol: Ate1^tm2.1Akas
Name: targeted mutation 2.1, Anna Kashina
MGI accession ID: MGI:4414839
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Synonyms: Ate1^f
Marker symbol: Ate1
Marker name: arginyltransferase 1
Marker synonyms: Ate1_1; Ate1_2; Ate1_3; Ate1_4
Chromosome: 7
Strain of origin: (129S6/SvEvTac x C57BL/6J)F1
Molecular note: A loxP site was inserted upstream of exon 1 and an frt flanked neo cassette with a 3' loxP site was inserted downstream of exon 3. Flp mediated recombination removed the neo cassette leaving exons 1 through 3 floxed.