STOCK Gata6^tm1Hadj/J

Stock number: 028096
Product name: Gata6H2B-Venus knock-in reporter
Research areas: Developmental Biology Research, Research Tools

Description

These Gata6^H2B-Venus knock-in reporter mice are suitable for use in applications related to live imaging and isolation of cells expressing Gata6, and the study of endoderm and mesoderm development.

Detailed description

The Gata6 gene encodes a zinc finger transcription factor that is important in heart and endoderm development. These knock-in/knock-out mice express Venus variant Yellow Fluorescent Protein from the endogenous Gata6 locus. Venus is detected in mesoderm and endoderm progenitor cells after endoderm differentiation in heterozygous embryos. Venus fluorescence is similar to endogenous Gata6 expression patterns. In heterozygous E8.25 embryos fluorescence is not detected in lateral mesoderm derived endocardium and body wall, but is detected in the yolk sac. In adult heterozygotes, fluorescence is detected in heart, lung, pancreas, liver, gall bladder, ovaries, adrenal glands, stomach (mucosa and mesothelium of the gastric corpus and small intestine), bladder, and skin (sebaceous glands). Mice that are heterozygous for the targeted mutation are viable, and may have reduced fertility (the number of adult heterozygotes are lower than the expected Mendelian ratio). Homozygotes are not viable. No Gata6 gene product (protein) is detected by immunostaining/immunofluorescence of homozygous E3.5 embryos.

Development

A targeting vector designed by Dr. Anna-Katerina Hadjantonakis (Memorial Sloan Kettering Cancer Center) containing (3' to 5'): FRT site, engrailed 2 intron and exon with splice acceptor site, H2B-Venus-polyadenylation sequence, rox site flanked beta actin promoter driven NEO cassette, loxP site flanked Gata6 exon 2, was used to disrupt intron 1 of the Gata6 gene, upstream of the translation initiation codons. The construct was electroporated into (C57BL/6 x 129S4/SvJae)F1 embryonic stem (ES) cells derived from KH2 ES cell mice carrying Gt(ROSA)26Sor^{tm1(rtTA*M2)Jae} and Col1a1^{tm13(neo/hygro*)Jae} alleles. (The KH2 ES cell mice were originally generated in (C57BL/6 x 129S4/SvJae)F1 derived v6.5 ES cells) Correctly targeted ES cells were injected into C2J blastocysts. The resulting chimeric males were crossed to CD-1 female mice. The Gata6^tm1Hadj (Gata6^H2B-Venus) allele was bred away from the other alleles through breeding to CD-1 females. 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).

Allele

Symbol: Gata6^tm1Hadj
Name: targeted mutation 1, Anna-Katerina Hadjantonakis
MGI accession ID: MGI:5700508
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Null/Knockout
Marker symbol: Gata6
Marker name: GATA binding protein 6
Chromosome: 18
Strain of origin: (C57BL/6 x 129S4/SvJae)F1
Expressed genes: YFP,Yellow Fluorescent Protein,jellyfish
Site of expression: Venus variant Yellow Fluorescent Protein is detected in mesoderm and endoderm progenitor cells.
Molecular note: The targeting vector is designed to insert downstream of the first non-coding intron with (from 3' to 5') an FRT site, engrailed 2 intron and exon with splice acceptor site, H2B-Venus-polyadenylation sequence, rox-flanked beta actin promoter driven NEO cassette, and a loxP site, followed by exon 2 and a second loxP site. Two alternative translation initiation codons are present in exon 2.
General note: The allele was made in KH2 cells that carry Gt(ROSA)26Sor^{tm1(rtTA*M2)Jae} and Col1a1^{tm2(tetO-Pou5f1)Jae} (J:159351).