C57BL/6-Gt(ROSA)26Sor^{tm1(HBEGF)Awai}/J

Stock number: 007900
Product name: ROSA26iDTR
Strain synonyms: B6-iDTR
Research areas: Neurobiology Research, Research Tools

Description

The Cre-inducible expression of DTR in these iDTR knock-in mice render cells susceptible to ablation following Diphtheria toxin administration.

Detailed description

Mice homozygous for this iDTR mutation are viable and fertile. These mice have the simian diphtheria toxin receptor (DTR; from simian Hbegf) inserted into the Gt(ROSA)26Sor (ROSA26) locus. Widespread expression of DTR is blocked by an upstream loxP-flanked STOP sequence. Of note, it has been the experience of The Jackson Laboratory that mice homozygous for the DTR, in the absence of Cre, are undersized at 3 weeks of age. We therefore recommend weaning these mice at ~4 weeks of age.

When bred to Cre recombinase-expressing mice, the STOP sequence is deleted in tissues where Cre is present, permitting DTR expression. Cells expressing DTR are rendered susceptible to ablation following Diphtheria toxin administration.

For example, when bred to a strain with a Cd19 null allele and expressing Cre recombinase during the B lymphocyte development (Stock No. 006785), this mutant mouse strain may be useful in studies of lymphocyte cell ablation.

When crossed to a strain expressing Cre recombinase in oocytes (see Stock No. 011062), this mutant mouse strain may be useful in studies of ovarian development.

When crossed to a strain expressing Cre recombinase in the pituitary and, at lower levels, in the testes (see Stock No. 011069), this mutant mouse strain may be useful in studies of metabolic dysfunction.

Of note, iDTR mice are also available on a BALB/cBy congenic background (as Stock No. 008040) and a NOD congenic background (as Stock No. 016603).

Of note, The Jackson Laboratory Repository also distributes RC::L-DTA mice (Stock No. 026944), which have Cre recombinase-dependent inverted tox176 attenuated diphtheria toxin subunit alpha gene (DTA*G128D), as well as widespread eGFP fluorescence in the absence of Cre recombinase.

Development

A targeting vector was designed with a loxP-flanked STOP cassette upstream of the open reading frame of the simian Diphtheria Toxin Receptor (DTR; from simian hbEGF cDNA (base pair 56-682)). The loxP-flanked region contains two SV40 polyA signals, an frt-flanked neomycin resistance gene, and a transcriptional STOP cassette. This construct was inserted into the Gt(ROSA)26Sor locus via electroporation of C57BL/6-derived Bruce4 ES embryonic stem (ES) cells. Correctly targeted ES cells were microinjected in CB20 blastocysts. Chimeric mice were bred to C57BL/6 to generate heterozygous "iDTR" mice. The donating investigator reported that these iDTR mice were maintained as homozygotes on the C57BL/6 genetic background (see SNP note below) prior to arrival at The Jackson Laboratory.

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 27 markers throughout the genome suggested a C57BL/6 genetic background, 3 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: Gt(ROSA)26Sor^{tm1(HBEGF)Awai}
Name: targeted mutation 1, Ari Waisman
MGI accession ID: MGI:3772576
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Inserted expressed sequence
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Chromosome: 6
Strain of origin: B6.Cg-Thy1^a
Expressed genes: HBEGF,heparin binding EGF like growth factor,chimpanzee; DTR,Simian Diphtheria Toxin Receptor,
Molecular note: A targeting vector was designed with a loxP-flanked STOP cassette upstream of the open reading frame of the simian Diphtheria Toxin Receptor (DTR; from simian hbEGF cDNA (base pair 56-682)). The loxP-flanked region contains two SV40 polyA signals, an frt-flanked neomycin resistance gene, and a transcriptional STOP cassette. This construct was inserted into the Gt(ROSA)26Sor locus via electroporation of C57BL/6-derived Bruce4 ES embryonic stem (ES) cells. Widespread expression of DTR is blocked by an upstream loxP-flanked STOP sequence. When bred to Cre recombinase-expressing mice, the STOP sequence is deleted in tissues where Cre is present, permitting DTR expression. Cells expressing DTR are rendered susceptible to ablation following Diphtheria toxin administration.