STOCK Gt(ROSA)26Sor^{tm1(birA)Mejr} Gata4^tm3.1Wtp/J

Stock number: 018121
Product name: Gata4^flbio Rosa26^bira
Research areas: Research Tools

Description

In these double mutant mice, GATA4 is biotinylated, allowing for specific imunoprecipitation of both GATA4 and the proteins GATA4 interacts with.

Detailed description

These biallelic Gata4^flbio mutant mice possess a FLAG/bio epitope tag downstream of the 3' UTR of the GATA binding protein 4 (Gata4) gene. GATA4 is a zinc-finger-containing transcription factor expressed in cardiomyocytes and has a role in regulating the expression of genes involved in cardiac differentiation. These mice also contain the Gt(ROSA)26Sor^{tm1.1(birA)Mejr} allele, which expresses HA (hemagglutinin) tagged bacterial birA, bifunctional protein (biotin ligase) (see Stock No. 010920). In these double mutant mice, GATA4 is biotinylated, allowing for specific imunoprecipitation of GATA4 and bound proteins. Homozygous mice are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities.

Development

A targeting vector was designed to insert a FLAG/bio epitope tag and a frt-flanked neomycin resistance (neo) cassette at the endogenous GATA binding protein 4 (Gata4) stop codon and upstream of the 3' UTR. The construct was electroporated into 129S4/SvJae-derived J1 embryonic stem (ES) cells and correctly targeted ES cells were injected into blastocysts. The resulting chimeric mice were bred to outbred Swiss Webster mice. Offspring were bred with Tg(ACTFLPe)9205Dym transgenic mice to delete the neo cassette. The resulting Gata4^flbio mice were maintained on a Swiss Webster background. These mice were subsequently bred to FVB;129P2-Gt(ROSA)26Sor^{tm1.1(birA)Mejr}/J mice (Stock No. 010920) and double homozygous mice were maintained by the Cardiovascular Development Consortium (CvDC), an NIH-funded consortium within the Bench-to-Bassinett (B2B) program. Upon arrival, mice were bred to FVB/NJ inbred mice (Stock No. 001800) for at least one generation to establish The Jackson Laboratory Repository colony.

Allele

Symbol: Gata4^tm3.1Wtp
Name: transgene insertion 3.1, William T Pu
MGI accession ID: MGI:5308206
Generation method: Targeted
Marker symbol: Gata4
Marker name: GATA binding protein 4
Chromosome: 14
Strain of origin: Not Specified
Molecular note: Flag and bio epitope tags and an FRT flanked neo cassette were inserted at the C terminus of the gene via homologous recombination. Flp mediated recombination removed the neo cassette.

Allele

Symbol: Gt(ROSA)26Sor^{tm1.1(birA)Mejr}
Name: targeted mutation 1.1, Dies Meijer
MGI accession ID: MGI:3588507
Generation method: Targeted
Attributes: Not Applicable
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Chromosome: 6
Strain of origin: 129P2/OlaHsd
Expressed genes: birA,biotin ligase),bacteria
Molecular note: An HA (hemagglutinin) tagged E. coli bacterial birA, bifunctional protein (biotin ligase), gene was inserted into exon 3 of the rabbit beta-globin gene. The entire birA ORF, with the rabbit beta globin splice acceptor and transcription termination sequences were excised to generate a targeting vector that also contained a floxed PGK- puromycin selection cassette. This targeting vector was inserted into the first intron of the Gt(ROSA)26Sor gene. Cre-mediated recombination removed the floxed puromycin cassette. Western blot of mutants demonstrated the expression of birA in all tissues tested, though levels of expression varied from tissue to tissue.