STOCK Gli3^tm1.1Amc/PsonJ

Stock number: 026135
Product name: Gli3^3xFLAG
Research areas: Research Tools

Description

These Gli3^3XFlag knockin mice contain an N-terminal 3XFLAG epitope tag and an AviTag biotinylation site inserted in the Gli3 (GLI-Kruppel family member GLI3) start site. These mice may be useful for understanding Sonic hedgehog signaling and identifying targets of Gli3 action in developing mouse limbs.

Detailed description

Gli3 (GLI-Kruppel family member GLI3) encodes a DNA-binding transcription factor and functions as a mediator of Sonic hedgehog signaling. Gli3^3XFlag knockin mice contain an N-terminal 3XFLAG epitope for use with anti-FLAG antibodies and an AviTag that allows for streptavidin capture. Mice homozygous for the mutation are viable and fertile with no apparent phenotype. Expression of Gli3 is not affected. These mice may be useful for understanding Sonic hedgehog signaling and identifying targets of Gli3 action in developing mouse limbs.

Development

The Gli3^3XFlag targeting vector, created in the laboratory of Dr. Andrew McMahon (University of Southern California), contains an N-terminal triple FLAG epitope tag inserted downstream of the ATG start site of the Gli3 (GLI-Kruppel family member GLI3) gene followed by sequence for an AviTag (GLNDIFEAQKIEWHE) and GSG linker sequence. A loxP-flanked neomycin cassette also is present in the construct. 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. Chimeric males were crossed to C57BL/6J females and progeny were crossed to B6.Cg-Edil3^{Tg(Sox2-cre)1Amc}/J to remove the neomycin cassette. Following an additional cross to C57BL/6J, mice were intercrossed and maintained as a homozygous colony. Dr. Kevin Peterson (The Jackson Laboratory) obtained homozygous Gli3^3XFlag mice in 2014 and cryorecovered using C57BL/6J donors. The resulting heterozygous animals were intercrossed to obtain homozygous colony.

Allele

Symbol: Gli3^tm1.1Amc
Name: targeted mutation 1.1, Andrew P McMahon
MGI accession ID: MGI:6457549
Generation method: Targeted
Attributes: Epitope tag
Synonyms: Gli3^3xFLAG
Marker symbol: Gli3
Marker name: GLI-Kruppel family member GLI3
Marker synonyms: ACLS; Bph; brachyphalangy; GCPS; GLI3-190; GLI3FL; PAPA; PAP-A; PAPA1; PAPB; PHS; PPDIV
Chromosome: 13
Strain of origin: (C57BL/6 x 129S4/SvJae)F1
Molecular note: Sequence for triple FLAG tags was inserted immediately downstream of the ATG translation start site, followed by sequence for an AviTag (GLNDIFEAQKIEWHE) and GSG linker sequence. The peptide encoded by this allele is thus tagged with N-terminal FLAG tags and contains an N-terminal biotinylation site that allows for streptavidin capture. The loxP site flanked neomycin resistance gene cassette that was also inserted was removed through subsequent cre-mediated recombination using mice carrying the