STOCK Gli1^tm2Alj/J

Stock number: 008211
Product name: Gli1^lz
Research areas: Developmental Biology Research, Neurobiology Research, Research Tools

Description

These lacZ reporter mutant mice harbor a β-galactosidase "knock-in" mutation that also abolishes endogenous GLI-Kruppel family member GLI1 (Gli1) gene function, and may be useful for studying Hedgehog/Sonic Hedgehog signaling in axis patterning, proliferation, and cell fate specification of Hedgehog responding cells at different stages of embryogenesis.

Detailed description

Mice homozygous for the Gli1^lz (or Gli1^lacZ) allele are viable and semi-fertile, with a "knock-in" of β-galactosidase (lacZ) inserted into the first coding exon (exon 2) and replacing the genomic fragment encoding the entire N-terminal and zinc-finger domains of the targeted locus (exons 2-7); abolishing endogenous gene function even if alternative splicing occurs. Under control of the upstream promoter/enhancer elements, lacZ expression is observed in a pattern indistinguishable from wildtype gene mRNA expression. As Gli1 transcription is a readout of high level Hedgehog signaling, these Gli1^lz (or Gli1^lacZ) mice may be useful for studying Hedgehog/Sonic Hedgehog signaling in axis patterning, proliferation, and cell fate specification of Hedgehog responding cells at different stages of embryogenesis.

Development

A targeting vector was designed to insert a nuclear localized β-galactosidase (lacZ) cDNA and loxP-flanked, reverse-oriented neo cassette into the first coding exon (exon 2) and delete the coding sequences in exons two to seven of the targeted gene. This construct was electroporated into 129S6/SvEvTac-derived W4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric mice were bred with 129SvEv (Taconic) mice. Next, mutant mice were bred to TK-Cre transgenic mice (129SvEv genetic background) to remove the floxed neo cassette. The resulting Gli1^lz (or Gli1^lacZ) offspring were selectively bred to remove the TK-Cre transgene and then subsequently maintained by breeding to Swiss Webster mice and also interbreeding for many generations prior to arrival at The Jackson Laboratory. Upon arrival, mutant mice were bred to 129S1/SvImJ (Stock No. 002448) for at least one generation to establish the colony.

Allele

Symbol: Gli1^tm2Alj
Name: targeted mutation 2, Alexandra L Joyner
MGI accession ID: MGI:2449767
Generation method: Targeted
Attributes: Reporter; Null/Knockout
Marker symbol: Gli1
Marker name: GLI-Kruppel family member GLI1
Chromosome: 10
Strain of origin: 129S6/SvEvTac
Expressed genes: lacZ,beta-galactosidase,E. coli
Site of expression: LacZ expression replaces that of the targeted gene in a pattern indistinguishable from that of the wildtype.
Molecular note: The gene was disrupted by replacement of a genomic region encoding the Gli N-terminal and zinc finger domains with a floxed-neo cassette and a lacZ having a nuclear localizing signal. The floxed-neo cassette was subsequently removed by crossing mutant animals to a cre deleter strain. The lacZ expression pattern in homozygous mutant embryos is identical to that of wild-type Gli as determined by immunohistochemical analysis.