These RCE:loxP mice harbor the R26R CAG-boosted EGFP (RCE) reporter allele with a loxP-flanked STOP cassette upstream of the enhanced green fluorescent protein (EGFP) gene. After removal of the loxP-flanked STOP cassette by cre-mediated recombination, EGFP reporter expression is directed to the cells/tissues dependent upon expression pattern of the promoter driving Cre recombinase.
Gord Fishell, NYU Neuroscience Institute
Mice homozygous for the R26R CAG-boosted EGFP (RCE) reporter allele harboring a loxP-flanked STOP cassette (RCE:loxP) 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 floxed-STOP cassette. After removal of the loxP-flanked STOP cassette by cre-mediated recombination, EGFP reporter expression is directed to the cells/tissues dependent upon expression of the promoter expression driving Cre 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 FLP-expressing strain (unspecified genetic background) to delete the FRT-flanked modified pBS302 STOP cassette. The resulting RCE:loxP offspring were mated with outbred Swiss Webster mice for many generations (selectively removing the FLP-expressing mutation), 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:loxP colony.
|Allele Name||targeted mutation 1.1, Gordon Fishell|
|Allele Type||Targeted (Reporter)|
|Allele Synonym(s)||RCE; RCE-GFP; RCE-LoxP; RCE:loxP; RCEEGFP; RCEfl; RCEfsEGFP|
|Gene Symbol and Name||Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano|
|Gene Synonym(s)||AV258896; Gtrgeo26; Gtrgeo26; Gtrosa26; Gtrosa26; R26; ROSA26; SETD5-AS1; beta geo; expressed sequence AV258896; gene trap ROSA 26; gene trap ROSA b-geo 26|
|Strain of Origin||129S6/SvEvTac|
|Mutations Made By|| |
Gord Fishell, NYU Neuroscience Institute
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