These RCE:FRT mice harbor the R26R CAG-boosted EGFP (RCE) reporter allele with a FRT-flanked STOP cassette upstream of the enhanced green fluorescent protein (EGFP) gene. After removal of the FRT-flanked STOP cassette by Flp-mediated recombination, EGFP reporter expression is directed to the cells/tissues dependent upon expression of the promoter driving FLP recombinase.
Gord Fishell, Harvard Medical School
Genetic Background | Generation |
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?+pN2F2
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Null/Knockout) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
Mice homozygous for the R26R CAG-boosted EGFP (RCE) reporter allele harboring an FRT-flanked STOP cassette (RCE:FRT) are viable and fertile. Under control of the endogenous Gt(ROSA)26Sor promoter/enhancer regions and the CMV-IE enhancer/chicken beta-actin/rabbit beta-globin (CAG) hybrid promoter, widespread expression of enhanced green fluorescent protein (EGFP) is prevented by the upstream FRT-flanked STOP cassette. After removal of the FRT-flanked STOP cassette by Flp-mediated recombination, EGFP reporter expression is directed to the cells/tissues dependent upon the expression pattern of the promoter driving FLP recombinase.
The R26R CAG-boosted EGFP (RCE) reporter targeting vector was designed with (from 5' to 3') the CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG; from the pCAGGS vector), a splice acceptor site, a loxP-flanked transcriptional STOP cassette (PGK-Neo-polyA and 3x-polyA), an FRT-flanked modified pBS302 STOP cassette, an enhanced green fluorescent protein (EGFP) sequence, and two AttP-polyA sequences. This entire construct was inserted into the first exon of the Gt(ROSA)26Sor locus via electroporation into 129S6/SvEvTac-derived W4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts, and chimeric mice were bred with C57BL/6 mice to generate the RCE:dual colony. RCE:dual mice were then bred with a germline Cre-expressing strain (unspecified genetic background) to delete the loxP-flanked transcriptional STOP cassette. The resulting RCE:FRT offspring were mated with outbred Swiss Webster mice for many generations (selectively removing the Cre-expressing transgene), and subsequently maintained as homozygotes prior to arrival at The Jackson Laboratory. Upon arrival, mutant mice were bred to C57BL/6J inbred mice for at least one generation to establish the RCE:FRT colony.
Allele Name | targeted mutation 1.2, Gordon Fishell |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | RCE:FRT |
Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano |
Gene Synonym(s) | |
Site of Expression | EGFP will be expressed in cells where promoters driving Flp recombinase are expressed. |
Strain of Origin | 129S6/SvEvTac |
Chromosome | 6 |
Molecular Note | Cre mediated recombination removes the loxP flanked STOP cassette from Gt(ROSA)26Sortm1(CAG-EGFP)Fsh. After removal of the FRT-flanked STOP cassette by Flp-mediated recombination, EGFP reporter expression is directed to the cells/tissues dependent upon the expression pattern of the promoter driving FLP recombinase. |
Mutations Made By | Gord Fishell, Harvard Medical School |
When maintaining a live colony, heterozygous mice may be bred together or to wildtype mice from the colony. Homozygous mice may also be bred together.
When using the RCE:FRT mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #32038 in your Materials and Methods section.
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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