C57BL/6J-Gabra2^{C57BL/6J+em1Geh}/J

Stock number: 034310
Product name: Gabra2^KI

Description

Gabra2^KI mice have a CRISPR/Cas9-generated thymine insertion at the intron/exon junction at 71,014,638 bp (GRCm38/mm10) of the gamma-aminobutyric acid (GABA) A receptor, subunit alpha 2 (Gabra2) gene. This mutation corrects a naturally occurring deletion found specifically in the C57BL/6J substrain.

Detailed description

CRISPR/cas9 endonuclease mediated genome editing of the gamma-aminobutyric acid (GABA) A receptor, subunit alpha 2 (Gabra2) gene was used to introduce a thymine residue at the intron/exon junction at 71,014,638 bp (GRCm38/mm10). The targeted gene encodes a subunit of GABA type-A (GABA-A) receptors that is expressed in the hippocampus, frontal cortex, amygdala, dorsal and ventral striatum, and hypothalamus and is important for motivation, reward, anxiety, depression, and fear. Mutations in GABRA2 have been implicated in epilepsy, affective and psychiatric disorders, alcoholism, and drug abuse. The C57BL6/J substrain has a spontaneous intronic deletion that results in a decrease of GABRA2 protein and mRNA expression. This insertion fully restores GABRA2 protein and mRNA levels in the brain tissue of B6-homozygous Gabra2^KI/KI and heterozygous Gabra2^KI/B6J F2 offspring.

Development

Plasmids encoding a guide RNA designed to introduce a T insertion at the intron/exon junction at 71,014,638 bp (GRCm38/mm10) of the gamma-aminobutyric acid (GABA) A receptor, subunit alpha 2 (Gabra2) gene and the cas9 nuclease were introduced into the cytoplasm of C57BL/6J-derived fertilized eggs with well recognized pronuclei. Correctly targeted embryos were transferred to pseudopregnant females. Pups were identified by sequencing and PCR and male founder (F0) was bred to a C57BL/6J female. Resulting Gabra2^KI offspring were intercrossed to develop the colony. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. 000664) oocytes.

Allele

Symbol: Gabra2^{C57BL/6J+em1Geh}
Name: endonuclease-mediated revertant 1, Gregg E Homanics
MGI accession ID: MGI:6381661
Generation method: Endonuclease-mediated
Attributes: Modified regulatory region
Synonyms: Gabra2^em1Geh; Gabra2^KI; Rr126^{C57BL/6J+em1Geh}
Marker symbol: Gabra2
Marker name: gamma-aminobutyric acid (GABA) A receptor, subunit alpha 2
Marker synonyms: C630048P16Rik; DEE78; EIEE78; Gabra-2
Chromosome: 5
Strain of origin: C57BL/6J
Molecular note: CRISPR/Cas9 was used to insert a single base pair (A on negative strand) between chr5:71171981-71171982 (GRCm39) to correct the spontaneous deletion of an A in intron 4 that is inherent in the C57BL/6J and BXD40 strains. This restores the allele to the "wild-type" form as found in C57BL/6NJ, C57BL/6ByJ, C57BL/6EiJ, DBA/2J and others.