B6(129S4)-Et(cre/ERT2)1645Rdav/J

Stock number: 009582
Strain synonyms: C57BL/6-Et(cre/ERT2)1645Rdav/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.

For characterization information, see images at the Allen Institute for Brain Science website (Et(cre/ERT2)1645Rdav 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 mice (F0) from line 1645 were bred to "C57 R26R" mutant mice from the Baylor College of Medicine (Gt(ROSA)26Sor^tm1Sor mice; similar to, but perhaps not as backcrossed as Stock No. 003474). The donating investigator stated that Cre-positive transgenic pups were bred to C57BL/6 mice or "C57 R26R" (see SNP note below) to maintain the colony. Upon arrival, transgenic mice were selectively bred to C57BL/6J inbred mice (Stock No. 000664) to remove the Gt(ROSA)26Sor^tm1Sor mutation.

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 26 of 27 markers throughout the genome suggested a C57BL/6 genetic background (1 suggested 129 background on Chromosome 13), at least of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.

Allele

Symbol: Et(cre/ERT2)1645Rdav
Name: enhancer trap 1645, Ron Davis
MGI accession ID: MGI:3852332
Generation method: Transgenic
Attributes: Recombinase-expressing; Inducible
Marker symbol: Et(cre/ERT2)1645Rdav
Marker name: enhancer trap 1645, 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 1645 was established on a C57BL/6J genetic background.