B6.Cg-Gt(ROSA)26Sor^{tm1(rtTA,EGFP)Nagy}/J

Stock number: 005670
Product name: ROSA26-rtTA-IRES-EGFP
Strain synonyms: rtTA^flox
Research areas: Research Tools

Description

ROSA26-rtTA mice can be used to generate triple mutant mice in which the tissue specificity of a cre-transgenic line and doxycycline inducibility of the rtTA/TRE-controlled transgenes can be combined to regulate expression of a target gene.

Detailed description

Homozygous mutant mice are viable, fertile, normal in size and do not display any behavioral abnormalities. When these gene targeted mice are bred to transgenic strains expressing Cre recombinase, functional rtTA and EGFP activity is observed in the double mutant offspring in the tissues that express cre. These double mutant mice may be bred to transgenic strains carrying genes of interest under the regulation of tetracycline responsive elements (TRE; tetO) to generate triple mutant mice in which the tissue specificity of the cre-transgenic line and doxycycline inducibility of the rtTA/TRE-controlled transgenes can be combined to regulate expression of the target gene.

In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. It should be noted that the phenotype could vary from that originally described. We will modify the strain description if necessary as published results become available.

Development

A targeting vector containing a loxP-flanked phosphoglycerate kinase-neomycin resistance gene-polyadenylation stop sequence (PGK-neo-pA) and a downstream reverse tetracycline-controlled transactivator gene-internal ribosomal entry site-enhanced green fluorescent protein gene-polyadenylation stop sequence (rtTA-IRES-EGFP-pA) fusion protein was inserted between exon 1 and 2 of the endogenous Gt(ROSA)26Sor locus. The construct was electroporated into (129X1/SvJ x 129S1/Sv)-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into ICR blastocysts and the resulting chimeric males were backcrossed for germ-line transmission to ICR females. Heterozygous offspring were sent to The Jackson Laboratory (as Stock No. 005572). Upon arrival, some mice were backcrossed to C57BL/6J for at least 5 generations to generate this congenic strain (Stock No. 005670).

Allele

Symbol: Gt(ROSA)26Sor^{tm1(rtTA,EGFP)Nagy}
Name: targeted mutation 1, Andras Nagy
MGI accession ID: MGI:3583817
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Transactivator
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Chromosome: 6
Strain of origin: (129X1/SvJ x 129S1/Sv)F1-Kitl⁺
Expressed genes: rtTA,reverse tetracycline-controlled transactivator,E. coli; GFP,Green Fluorescent Protein,
Site of expression: When these mutant mice are crossed to a strain expressing Cre recombinase, rtTA and EGFP are expressed in the cre-expressing tissues. These mice can be used to generate mutant mice in which the tissue specificity of cre-expression and doxycycline inducibility of the rtTA can be combined.
Molecular note: A targeting vector containing a floxed Pgk-neo-pA cassette and a rtTA-IRES-EGFP-pA cassette was inserted into intron 1 of the locus. Upon cre expression, the floxed neo cassette is removed and the ROSA26 promoter drives expression of rtTA and EGFP. Presence of doxycycline results in the formation of an active transcriptional activator and the activation of the responder transgene.