B6.Cg-Tg(GFAP-cre/ERT2)505Fmv/J

Stock number: 012849
Product name: GCE
Research areas: Neurobiology Research, Research Tools

Description

These GCE transgenic mice have a human glial fibrillary acidic protein (hGFAP) promoter sequence directing expression of a Cre-estrogen receptor (Cre-ERT2) fusion protein. When these transgenic mice are bred with mice containing a loxP-flanked sequence, tamoxifen-inducible Cre-mediated recombination is expected to result in deletion of the floxed sequences in the Cre recombinase-expressing tissues of the offspring.

Detailed description

Mice hemizygous for the GCE transgene are viable and fertile, with a human glial fibrillary acidic protein (hGFAP) promoter sequence directing expression of a Cre-estrogen receptor (Cre-ERT2) fusion protein. This Cre-ERT2 fusion protein is only active when it binds to the estrogen analog 4-hydroxytamoxifen (OHT). Specifically, Cre recombinase expression is observed in subventricular zone (SVZ), dentate gyrus (DG), celebellum, mesencephalon, diencephalon, and cerebral cortex. Cre expression was also enriched in the sub-granular zone of the DG and the cerebellar Purkinje layer. When these transgenic mice are bred with mice containing a loxP-flanked sequence, tamoxifen-inducible Cre-mediated recombination is expected to result in deletion of the floxed sequences in the Cre recombinase-expressing tissues of the offspring.

The Cre-ERT2 fusion protein consists of Cre recombinase fused to a triple mutant form of the human estrogen receptor which does not bind its natural ligand (17β-estradiol) at physiological concentrations but will bind the synthetic estrogen receptor ligands 4-hydroxytamoxifen (OHT or tamoxifen) and, with lesser sensitivity, ICI 182780. Restricted to the cytoplasm, Cre-ERT2 can only gain access to the nuclear compartment after exposure to tamoxifen. To counteract the mixed estrogen agonist effects of tamoxifen injections, which can result in late fetal abortions in pregnant mice, progesterone may be coadministered.

Development

The GCE transgene was designed containing a Cre-estrogen receptor (Cre-ER^T2) fusion protein (Cre-ERT2; Cre recombinase fused to a G400V/M543A/L544A triple mutation of the human estrogen receptor ligand binding domain) driven by the human glial fibrillary acidic protein (hGFAP) promoter. The transgene was microinjected into fertilized (C57BL/6J x SJL/J)F2 oocytes, and founder mice from line 505 were bred to C57BL/6J mice. Subsequently these mice were backcrossed to C57BL/6J mice for at least 8 generations to establish a colony of GCE mice. Upon arrival at The Jackson Laboratory, transgenic mice were bred to C57BL/6J inbred mice (Stock No. 000664) to establish the colony.

Allele

Symbol: Tg(GFAP-cre/ERT2)505Fmv
Name: transgene insertion 505, Flora M Vaccarino
MGI accession ID: MGI:3774167
Generation method: Transgenic
Attributes: Recombinase-expressing; Inducible
Marker symbol: Tg(GFAP-cre/ERT2)505Fmv
Marker name: transgene insertion 505, Flora M Vaccarino
Chromosome: UN
Strain of origin: (C57BL/6 x SJL/J)F2
Expressed genes: cre,cre recombinase,bacteriophage P1
Site of expression: Cre recombinase expression is observed in subventricular zone (SVZ), dentate gyrus (DG), celebellum, mesencephalon, diencephalon, and cerebral cortex. Cre expression was also enriched in the sub-granular zone of the DG and the cerebellar Purkinje layer.
Molecular note: The human GFAP promoter drives the inducible expression of a cre-ERT2 fusion cassette.