STOCK Gabrg2^tm1Wul/J

Stock number: 021197
Research areas: Neurobiology Research, Research Tools

Description

These γ2I77lox mice contain the point mutation F77I and contain loxP sites flanking exon 4 of the gamma-aminobutyric acid (GABA) A receptor, subunit gamma 2 (Gabrg2) gene. GABAA receptors are rendered zolpidem-insensitive and allow cell type-specific ablations of the GABAA receptor γ2 subunit.

Development

A targeting vector was designed to insert a loxP site upstream of exon 4 followed by a frt-flanked neomycin resistance (neo) cassette, and a second loxP site downstream of exon 4 of the gamma-aminobutyric acid (GABA) A receptor, subunit gamma 2 (Gabrg2) gene. A point mutation, resulting in a phenylalanine (F) to isoleucine (I) substitution at amino acid position 77, was introduced into exon 4. The construct was electroporated into (129X1/SvJ x 129S1/Sv)F1-Kitl⁺-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and resulting chimeric mice were bred with B6.Cg-Tg(ACTFLPe)9205Dym/J transgenic mice (Stock No. 005703) to delete the neo cassette. Subsequently, these γ2I77lox mice were bred to C57BL/6J mice for two generations to remove the Flp-expressing transgene. These mice were maintained on a mixed background and also contained the Hdc^tm1(cre)Wwis allele, which was removed prior to shipping. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation.

Allele

Symbol: Gabrg2^tm1Wul
Name: targeted mutation 1, Peter Wulff
MGI accession ID: MGI:3721943
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Gabrg2
Marker name: gamma-aminobutyric acid (GABA) A receptor, subunit gamma 2
Chromosome: 11
Strain of origin: (129X1/SvJ x 129S1/Sv)F1-Kitl⁺
Molecular note: Exon 4 was replaced with an exon containing a phenylalanine to isoleucine mutation at position 77 (F77I). This point mutation confers drug insensitivity to zolpidem. A loxP site was inserted in the upstream intron and a frt-flanked neomycin cassette followed by another loxP site was inserted in the downstream intron. Chimeric mice were mated with a Flp deleter strain to remove the neomycin cassette. Genomic sequencing of homozygous mice confirmed the presence of the F to I mutation.