mT/mG mice (Stock No. 007676) have the same allele as Stock No. 007576 but has a C57BL/6J congenic background. ROSAmT/mG is a cell membrane-targeted, two-color fluorescent Cre-reporter allele. Prior to Cre recombination, cell membrane-localized tdTomato (mT) fluorescence expression is widespread in cells/tissues. Cre recombinase expressing cells (and future cell lineages derived from these cells) have cell membrane-localized EGFP (mG) fluorescence expression replacing the red fluorescence. ROSAmT/mG mice (Stock No. 007576 and Stock No. 007676) function similarly to ROSAnT-nG mice (Stock No. 023035 and Stock No. 023537), with ROSAmT/mG directed to cell membrane and ROSAnT-nG directed to nuclei.
Liqun Luo, Stanford University
Genetic Background | Generation |
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N11F12
|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed), Reporter) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
Mice homozygous for this mT/mG mutation are viable and fertile. These mice possess loxP sites on either side of a membrane-targeted tdTomato (mT) cassette and express strong red fluorescence in all tissues and cell types examined. Tail or whole body epifluorescence is sufficient to identify mT/mG mice from wildtype mice. When bred to Cre recombinase expressing mice, the resulting offspring have the mT cassette deleted in the cre expressing tissue(s), allowing expression of the membrane-targeted EGFP (mG) cassette located just downstream. The donating investigator reports that the ACTB promoter allows stronger and persistent expression of the fluorescent proteins (especially in adult cells) compared to the endogenous Gt(ROSA) locus alone. This double-fluorescent system allows direct live visualization of both recombined and non-recombined cells at single cell resolution, offering an internal control for phenotypic analysis of Cre-induced mosaic mutants and providing a second marker for lineage tracing applications. In addition, the localization of fluorescent proteins to membrane structures outlines cell morphology and allows resolution of fine cellular processes. These mT/mG mice are useful as a Cre reporter strain; expressing red fluorescence prior to, and green fluorescence following, Cre-mediated recombination in widespread cell and tissue types.
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.
The mT/mG (membrane-Tomato/membrane-Green) targeting vector was designed with a CMV enhancer/chicken beta-actin core promoter (pCA) driving expression of a loxP-flanked, N-terminal membrane-tagged, tdTomato protein sequence followed by a polyadenylation signal (tdTomato is a non-oligomerizing DsRed variant with a 12 residue linker fusing two copies of the protein (tandem dimer)). Immediately distal to the second loxP site is an N-terminal membrane-tagged, enhanced green fluorescent protein (EGFP) sequence itself followed by a polyadenylation signal. An frt-flanked neo cassette was also located distal to the expression vector. This entire mT/mG construct was inserted into the Gt(ROSA)26Sor locus via electroporation of (129X1/SvJ x 129S1/Sv)F1-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were microinjected in C57BL/6J blastocysts. Chimeric progeny were bred to outbred CD-1 mice. The resulting mutant mice were interbred to generate homozygotes prior to arrival at The Jackson Laboratory (as Stock No. 007576). Upon arrival, some mice were backcrossed to C57BL/6J for at least five generations to generate this congenic strain (Stock No. 007676).
Expressed Gene | GFP, Green Fluorescent Protein, |
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Expressed Gene | RFP, Red Fluorescent Protein, coral |
Site of Expression | Membrane-targeted tdTomato (mT) is expressed in all tissues and cell types examined. When bred to Cre recombinase expressing mice, the mT cassette is deleted in the cre expressing tissue(s), allowing expression of the membrane-targeted EGFP (mG). |
Allele Name | targeted mutation 4, Liqun Luo |
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Allele Type | Targeted (Conditional ready (e.g. floxed), Reporter) |
Allele Synonym(s) | Gt(ROSA26)ACTB-tdTomato-EGFP; mT/mG; mTmG; mTomato/mEGFP; R26TG; R26-mR/mG; R26-X; ROSAmT-mE; RosamTmG; Rosa26flox-mTRed-Stop-flox-mGFP; Rosa26mTmG; ROSA26-mTmG; Rosa26mTmGflox; Rosa26RmTmG; ROSA-mT/mG; ROSA-R/G; TomatoGFP+ |
Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano |
Gene Synonym(s) | |
Expressed Gene | GFP, Green Fluorescent Protein, |
Expressed Gene | RFP, Red Fluorescent Protein, coral |
Site of Expression | Membrane-targeted tdTomato (mT) is expressed in all tissues and cell types examined. When bred to Cre recombinase expressing mice, the mT cassette is deleted in the cre expressing tissue(s), allowing expression of the membrane-targeted EGFP (mG). |
Strain of Origin | (129X1/SvJ x 129S1/Sv)F1-Kitl+ |
Chromosome | 6 |
Molecular Note | The targeting vector was designed with a CMV enhancer/chicken beta-actin core promoter (pCA) driving expression of a loxP-flanked, N-terminal membrane-tagged, optimized DsRed fluorescent protein variant (called tandem-dimer-Tomato or tdTomato) sequence followed by a polyadenylation signal. Immediately distal to the second loxP site is an N-terminal membrane-tagged, enhanced green fluorescent protein (EGFP) sequence itself followed by a polyadenylation signal. An frt-flanked neo cassette was also located distal to the expression vector. Red fluorescence is detected in all tissues tested. When mice are bred to Cre expressing mice, the floxed region is excised in Cre-expressing tissue and this allows expression of the EGFP cassette. |
Mutations Made By | Liqun Luo, Stanford University |
Mutant mice were bred to C57BL/6J mice to generate this congenic strain. When maintaining the live congenic colony, homozygous mice may be bred together.
When using the mT/mG , mTmG mouse strain in a publication, please cite the originating article(s) and include JAX stock #007676 in your Materials and Methods section.
Service/Product | Description | Price |
---|---|---|
Heterozygous for Gt(ROSA)26Sor<tm4(ACTB-tdTomato,-EGFP)Luo> |
Frozen Mouse Embryo | B6.129(Cg)-Gt(ROSA)26Sor<tm4(ACTB-tdTomato -EGFP)Luo>/J Froz | $2595.00 |
Frozen Mouse Embryo | B6.129(Cg)-Gt(ROSA)26Sor<tm4(ACTB-tdTomato -EGFP)Luo>/J Froz | $2595.00 |
Frozen Mouse Embryo | B6.129(Cg)-Gt(ROSA)26Sor<tm4(ACTB-tdTomato -EGFP)Luo>/J Froz | $3373.50 |
Frozen Mouse Embryo | B6.129(Cg)-Gt(ROSA)26Sor<tm4(ACTB-tdTomato -EGFP)Luo>/J Froz | $3373.50 |
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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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