STOCK Slc32a1^tm2(cre)Lowl/J

Stock number: 016962
Product name: Vgat-ires-cre
Research areas: Cancer Research, Neurobiology Research, Research Tools

Description

A congenic version of this strain is available as Stock No. 028862.

Vgat-ires-Cre knock-in mice have Cre recombinase expression directed to inhibitory GABAergic neuron cell bodies, without disrupting endogenous vesicular inhibitory amino acid transporter expression. These mice may be useful for removing floxed sequences in studies of GABAergic neuronal function.
Of note, the same Vgat-ires-Cre knock-in allele is also available on a C57BL/6J genetic background as Stock No. 028862.

Detailed description

Slc32a1^tm2(cre)Lowl knock-in mice (commonly referred to as Vgat-ires-Cre mice) have Cre recombinase expression directed to inhibitory GABAergic neuron cell bodies, without disrupting endogenous vesicular inhibitory amino acid transporter expression. Both heterozygous and homozygous mice are viable and fertile with no gross physical abnormalities. Specifically, Cre recombinase activity is detected in inhibitory GABAergic neuron cell bodies in the caudate putamen, suprachiasmatic nucleus, central amygdaloid nucleus, and zona incerta, nucleus accumbens, lateral septum, medial septum, reticular nucleus of the thalamus, substantia nigra pars reticulata, and Purkinje cell layer of the cerebellum. When crossed with a strain containing loxP site-flanked sequences, Cre-mediated recombination results in tissue-specific deletion of flanked sequences in the offspring.

Of note, the targeted Slc32a1 gene is located ~3.5 kbp (~1.9 cM) from the nonagouti locus (A). These C57BL/6;FVB;129S6 Vgat-ires-Cre knock-in mice have agouti coat color; presumably due to the presence of 129S6-derived A^w allele (from the ES cells used in creating the Vgat-ires-Cre allele).

Development

A targeting vector was used to insert an IRES-Cre cassette downstream of the stop codon of the solute carrier family 32 (GABA vesicular transporter), member 1 gene (Slc32a1 or VGAT) on chromosome 2. The construct was electroporated into 129S6/SvEvTac derived W4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were crossed to C57BL/6 mice, and then crossed to FVB. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.

Allele

Symbol: Slc32a1^tm2(cre)Lowl
Name: targeted mutation 2, Bradford B Lowell
MGI accession ID: MGI:5141270
Generation method: Targeted
Attributes: Recombinase-expressing
Marker symbol: Slc32a1
Marker name: solute carrier family 32 (GABA vesicular transporter), member 1
Chromosome: 2
Strain of origin: 129S6/SvEvTac
Expressed genes: cre,cre recombinase,bacteriophage P1
Site of expression: Cre recombinase activity is detected in inhibitory GABAergic neuron cell bodies in the caudate putamen, suprachiasmatic nucleus, central amygdaloid nucleus, and zona incerta, nucleus accumbens, lateral septum, medial septum, reticular nucleus of the thalamus, substantia nigra pars reticulata, and Purkinje cell layer of the cerebellum.
Molecular note: A targeting vector, constructed from a BAC clone containing the Slc32a1 (Vgat) gene, was utilized to insert an internal ribosomal entry site followed by the cre recombinase sequence into the Slc32a1 locus just downstream of the endogenous stop codon. Cre is expressed from the Slc32a1 gene promoter. Immunohistochemistry on brain and Western blot analysis shows decreased expression of SLC32A1 protein in homozygotes.