C57BL/6-Gt(ROSA)26Sor^{tm1(CAG-FOXO1,GFP)Moli}/J

Stock number: 029316
Product name: Foxo1^AAA
Strain synonyms: R26StopFlFoxo1AAA
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

Foxo1^AAA knock-in mice express a Cre recombinase-dependent haemagglutinin-tagged mutant human FOXO1, and is useful in applications related to regulatory T cell differentiation, homeostasis and function.

Detailed description

The FOXO1 gene encodes a forkhead transcription factor that is important in metabolic homeostasis, cell cycle regulation, apoptosis, T-cell regulation and oxidative stress response. These Foxo1^AAA knock-in mice have Cre recombinase-dependent expression of a HA-tagged human FOXO1 (AAA) mutant cDNA from the Gt(ROSA)26Sor locus. A loxP-flanked ‘neo-STOP’ cassette prevents expression of the downstream mutant human FOXO1. The FOXO1 (AAA) mutant allele carries alanine amino acid substitutions at the Akt phosphorylation sites, resulting in defective nuclear export of the transcription factor.

When these mutant mice are bred to mice that express Cre recombinase, the resulting offspring will express HA-tagged mutant AAA human FOXO1 in the cre-expressing tissues. Mice that are homozygous for the targeted mutation are viable and fertile.

Development

A targeting vector containing CAG promoter, loxP-flanked Neo-STOP cassette, HA-tagged human FOXO1 (AAA) mutant cDNA, an FRT-flanked IRES/EGFP, and polyadenylation sequence, was inserted into the Gt(ROSA)26Sor locus. The construct was electroporated into unspecified C57BL/6-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were bred to C57BL/6 mice to test for germline transmission. The mice were maintained on the C57BL/6 background (see SNP note below). Upon arrival, 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. One the 43 markers throughout the genome was segregating, also 3 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.

Allele

Symbol: Gt(ROSA)26Sor^{tm1(CAG-FOXO1,GFP)Moli}
Name: targeted mutation 1, Ming O Li
MGI accession ID: MGI:5447989
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence; Humanized sequence
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Chromosome: 6
Strain of origin: C57BL/6
Expressed genes: Foxo1,forkhead box O1,mouse, laboratory; GFP,Green Fluorescent Protein,; HA,influenza hemagglutinin,
Site of expression: When these mutant mice are bred to mice that express Cre recombinase, the resulting offspring will express HA-tagged mutant AAA human FOXO1 in the cre-expressing tissues.
Molecular note: The targeting contruct inserted into intron 1 contained a CAG promoter, floxed neo/STOP cassette, HA-tagged human cDNA, an FRT-flanked IRES/EGFP and a polyadenylation sequence.