STOCK Gt(ROSA)26Sor^tm3(Gli3)Amc/J

Stock number: 013124
Research areas: Developmental Biology Research, Internal/Organ Research, Research Tools

Description

These RosaGli3TFlag c/c mice contain a floxed-neomycin resistance (neo) cassette and polyadenylation signal, cDNA encoding a FLAG-tagged GLI-Kruppel family member (Gli3) repressor gene, an internal ribosome entry site (IRES), and a Venus yellow fluorescent protein (YFP) under control of the ubiquitous Gt(ROSA)26Sor locus. These mice may be useful for understanding Sonic hedgehog signaling and identifying targets of Gli3 action in developing mouse limbs.

Detailed description

These RosaGli3TFlag c/c mice contain a floxed-neomycin resistance (neo) cassette and polyadenylation signal, cDNA encoding a FLAG-tagged GLI-Kruppel family member (Gli3) repressor gene, an internal ribosome entry site (IRES), and a Venus yellow fluorescent protein (YFP) under control of the ubiquitous Gt(ROSA)26Sor locus. Breeding these mutant mice to mice that express Cre-recombinase will also result in Floxed-neo-stop excision. When these mice are crossed to mice containing Cre-recombinase under direction of a paired related homeobox 1 (Prrx1) promoter (see Stock No. 005584), active in early limb mesenchyme, the mice produce Gli3TFlag at levels that are comparable with the endogenous protein. Mice exhibited a variety of limb defects including a variable preaxial forelimb polydactyly, limb truncation, and reduced mineralization. These mice may be useful for understanding Sonic hedgehog signaling and identifying targets of Gli3 action in developing mouse limbs.

Development

A targeting vector was designed to contain (from 5' to 3) a floxed-neomycin resistance (neo) cassette and polyadenylation signal, cDNA encoding a FLAG-tagged GLI-Kruppel family member (Gli3) gene, an internal ribosome entry site (IRES), and a Venus yellow fluorescent protein (YFP) under control of the endogenous Gt(ROSA)26Sor locus. The construct was electroporated into 129-derived YFP3.1 embryonic stem (ES) cells. These ES cells carry the tamoxifen inducible CAGGS-CreERTM transgene and are heterozygous at the Gt(ROSA)26Sor locus (Gt(ROSA)26Sor^tm1Sor/(ROSA)26Sor^tm1(EYFP)Cos). Correctly targeted ES cells were injected into C57BL/6 blastocysts and the resulting chimeric males were bred to C57BL/6 females to generate a colony of RosaGli3T FLAG c/c mice. These mice were subsequently backcrossed 2 generations to C57BL/6 background. These mice no longer carry the cre transgene. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J (Stock No. 000664) for at least one generation to establish the colony.

Allele

Symbol: Gt(ROSA)26Sor^tm3(Gli3)Amc
Name: targeted mutation 3, Andrew P McMahon
MGI accession ID: MGI:3828280
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Chromosome: 6
Strain of origin: 129 and C57BL/6 and CBA and Swiss Webster
Expressed genes: Gli3,GLI-Kruppel family member GLI3,mouse, laboratory
Molecular note: A loxP site flanked neomycin resistance gene cassette, a cDNA encoding the first 740 amino acids of Gli3, three copies of the sequence coding for a flag tag, and an IRES-YFP (Venus) cassette were inserted into the locus. Expression is only achieved following cre-mediated removal of the floxed neo cassette.
General note: ES cell line = YFP3.1, a male ES cell line carrying Tg(CAG-cre/Esr1*)5Amc and is compound heterozgous at Gt(ROSA)26Sor: Gt(ROSA)26Sor^tm1Sor and Gt(ROSA)26Sor^tm1(EYFP)Cos.