B6.Cg-Gt(ROSA)26Sor^{tm1(tetO-Yap1*,-GFP*)Stang}/J

Stock number: 031279
Product name: tetO-YAP-GFP
Research areas: Developmental Biology Research, Endocrine Deficiency Research, Research Tools

Description

Expression of a mutant S112A YAP1 protein and the nuclear-targeted fluorescent reporter HIST1H2BJ/GFP (H2B-GFP) is regulated by the Gt(ROSA)26Sor-driven tetracycline-responsive regulatory element (tetO) in these mice. When mated with a strain expressing tetracycline-controlled transactivator protein (tTA), expression of the mutant S112A YAP1 protein and HIST1H2BJ/GFP may be regulated with the tetracycline analog doxycycline (dox) in the double-allelic offspring. This strain may be useful in studies related to Hippo pathway signaling, acinar and endocrine tissue differentiation.

Detailed description

The Yap1 gene encodes a downstream effector of the Hippo pathway. Mutations in the Yap1 gene have been associated with ocular coloboma, with or without hearing impairment, cleft lip/palate, and/or mental retardation. These knock-in mice express a mutant Yap1, and the nuclear-targeted fluorescent reporter HIST1H2BJ/GFP (H2B-GFP) under the regulatory control of a Gt(ROSA)26Sor-driven tetO promoter. The mutant Yap1 contains an amino acid substitution of serine-to-alanine at position 112 (S112A), which confers enhanced nuclear translocation and activation of the YAP protein.
The Jackson Laboratory colony was viable and fertile as heterozygous or homozygous, but breeding homozygotes together was less productive than breeding heterozygotes.

When mated to a mutant strain expressing tetracycline-controlled transactivator protein (tTA), expression of the mutant S112A YAP1 protein and H2B-GFP (HIST1H2BJ/GFP) may be regulated with tetracycline or its analog doxycycline (dox) in the double mutant offspring. When crossed to a reverse tetracycline transactivator reporter strain, doxycycline-inducible expression of the mutant YAP1 and GFP is detected in pancreatic epithelial cells and some mesenchymal cells.

Development

A targeting vector containing a loxP site flanked PGK-neo cassette, a mutant mouse Yap1 gene (with an amino acid substitution of serine-to-alanine at position 112), viral 2A peptide sequence, HIST1H2BJ/GFP (H2B-GFP) cDNA, beta-globin and polyadenylation sequence, under the control of the tetO, tetracycline-responsive regulatory element (TRE-tight), was inserted into the Gt(ROSA)26Sor locus. The construct was electroporated into (C57BL/6 x 129S4/SvJae)F1 derived V6.5 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission. The mice were crossed to CD1 mice, and then backcrossed to C57BL/6 for 11 generations. 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: Gt(ROSA)26Sor^{tm1(tetO-Yap1*,-GFP*)Stang}
Name: targeted mutation 1, Ben Stanger
MGI accession ID: MGI:6115248
Generation method: Targeted
Attributes: Reporter; Inducible; Inserted expressed sequence
Synonyms: tetO-YAP-GFP
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Marker synonyms: beta geo; Gtrgeo26; Gtrosa26; R26; ROSA26; SETD5-AS1; Thumpd3as1
Chromosome: 6
Strain of origin: (C57BL/6 x 129S4/SvJae)F1
Expressed genes: Yap1,yes-associated protein 1,mouse, laboratory; GFP,Green Fluorescent Protein,
Site of expression: When mated to a mutant strain expressing tTA, expression of the mutant YAP1 protein and GFP may be regulated with tetracycline or its analog doxycycline in the double mutant offspring. When crossed to a rtTA reporter strain, doxycycline-inducible expression of the mutant YAP1 and GFP is detected in pancreatic epithelial cells and some mesenchymal cells.
Molecular note: The targeting vector contains a loxP site flanked PGK-neo cassette, a mutant mouse Yap1 gene (with an amino acid substitution of serine-to-alanine at position 112), viral 2A peptide sequence, HIST1H2BJ/GFP (H2B-GFP) cDNA, beta-globin and polyadenylation sequence, under the control of the tetO, tetracycline-responsive regulatory element (TRE-tight).