B6(129S4)-Et(cre/ERT2)398Rdav/J

Stock number: 009578
Strain synonyms: C57BL/6-Et(cre/ERT2)398Rdav/J
Research areas: Neurobiology Research, Research Tools

Description

These mice express the 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 (although some cre activity is reported prior to 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, 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)398Rdav 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 UAS-TK-cre:ERT2-bGHpA transgene was designed with an upstream activation sequence, a thymidine kinase minimal promoter, 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 398 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). Cre-positive transgenic pups were bred to C57BL/6 mice or "C57 R26R" 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.

In March 2010, a single nucleotide polymorphism (SNP) assay predicted these mice were 94% congenic onto the C57BL/6 genetic background (8 of 141 markers were not C57BL/6-allele type (on chr. 6, 9, 11, and 14).

Allele

Symbol: Et(cre/ERT2)398Rdav
Name: enhancer trap 398, Ron Davis
MGI accession ID: MGI:3852327
Generation method: Transgenic
Attributes: Recombinase-expressing; Inducible
Marker symbol: Et(cre/ERT2)398Rdav
Marker name: enhancer trap 398, 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 (although some cre activity is reported prior to tamoxifen administration).
Molecular note: An enhancer trap transgenic approach was used to generate these mice. The TK-bgi-cre:ERT2-bGHpA transgene was designed with the thymidine kinase (TK) 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). The bovine growth hormone polyadenylation signal follows the cre sequence. This transgene was injected into C57BL/6J embryos. Founder line 296 was established on a C57BL/6J genetic background.