B6(129S4)-Et(cre/ERT2)7089Rdav/J

Stock number: 010690
Strain synonyms: C57BL/6-Et(cre/ERT2)7089Rdav/J
Research areas: Neurobiology Research, Research Tools

Description

These mice express the tamoxifen-inducible Cre-ERT2 fusion protein (Cre-ER^T2) under the control of the promoter/enhancer regions surrounding the site where the enhancer trap transgene randomly inserted. These mice may be useful for generating conditional mutations for studying gain or loss of function and/or fate mapping in brain tissues.

Detailed description

Mice hemizygous for this enhancer trap transgene are viable and fertile, with expression of the Cre-ERT2 fusion gene under control of the promoter/enhancer regions surrounding the site where the enhancer trap transgene randomly inserted. Cre recombinase activity is observed in brain tissues following tamoxifen administration. The donating investigators may not have assessed expression in tissues other than brain. When these enhancer trap transgenic mice are bred with mice containing a loxP-flanked sequence of interest, tamoxifen-inducible, Cre-mediated recombination will result in deletion of the floxed sequence(s) in cre-expressing tissues in the offspring.

Variable expression of a reporter gene has been reported in the feet, ears, and tail prior to induction of cre gene expression.

For characterization information, see images at the Allen Institute for Brain Science website (Et(cre/ERT2)7089Rdav images).

The Cre-ERT2 fusion protein (Cre-ER^T2) 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

An enhancer trap transgenic approach was used to generate these mice. The hsyn-bgi-Cre:ErT2-bGHpA transgene was designed with a human synapsin minimal promoter, a beta-globin intron, a CreERT2 fusion gene (Cre-ER^T2; Cre recombinase fused to a G400V/M543A/L544A triple mutation of the human estrogen receptor ligand binding domain), and a bovine growth hormone polyA signal. This transgene was injected into C57BL/6J embryos. Founder line 7089 was established on a C57BL/6J genetic background. Transgenic mice were subsequently bred for multiple generations with C57BL/6J mice and/or with Gt(ROSA)26Sor^tm1Sor mutant mice (backcrossed to C57BL/6 for at least 10 generations) prior to arrival at The Jackson Laboratory. Upon arrival, enhancer trap transgenic mice were selectively bred to C57BL/6J inbred mice (Stock No. 000664) to remove the Gt(ROSA)26Sor^tm1Sor mutation.

SNP (single nucleotide polymorphism) analysis performed by The Jackson Laboratory revealed that the Donating Investigator used C57BL/6N (5 of 5 markers segregating) in the development of the strain. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J at least once to establish the colony.

Allele

Symbol: Et(cre/ERT2)7089Rdav
Name: enhancer trap 7089, Ron Davis
MGI accession ID: MGI:3852340
Generation method: Transgenic
Attributes: Recombinase-expressing; Inducible
Marker symbol: Et(cre/ERT2)7089Rdav
Marker name: enhancer trap 7089, Ron Davis
Chromosome: UN
Strain of origin: C57BL/6J
Expressed genes: cre/ERT2,Cre recombinase and estrogen receptor 1 (human) fusion gene,
Site of expression: Cre recombinase activity is observed in brain tissues following tamoxifen administration.
Molecular note: An enhancer trap transgenic approach was used to generate these mice. The hsyn-bgi-cre:ERT2-bGHpA transgene was designed with the human synapsin minimal promoter (hsyn) minimal promoter driving a cre-ER^T2 fusion gene (cre recombinase fused to a G400V/M543A/L544A triple mutation of the human estrogen receptor ligand binding domain).A beta globin intron separates the promoter from the cre gene and the bovine growth hormone polyadenylation signal follows the cre sequence. This transgene was injected into C57BL/6J embryos. Founder line 7089 was established on a C57BL/6J genetic background.