Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus

By S. Srinivas, Tomoko Watanabe, Chyuan-Sheng Lin, C. William, Y. Tanabe, T. Jessell, F. Costantini. BMC Developmental Biology, 2001

3,190 citations recorded by the source corpus.

BackgroundSeveral Cre reporter strains of mice have been described, in which a lacZ gene is turned on in cells expressing Cre recombinase, as well as their daughter cells, following Cre-mediated excision of a loxP-flanked transcriptional "stop" sequence. These mice are useful for cell lineage tracing experiments as well as for monitoring the expression of Cre transgenes. The green fluorescent protein (GFP) and variants such as EYFP and ECFP offer an advantage over lacZ as a reporter, in that they can be easily visualized without recourse to the vital substrates required to visualize β-gal in living tissue.ResultsIn view of the general utility of targeting the ubiquitously expressed ROSA26 locus, we constructed a generic ROSA26 targeting vector. We then generated two reporter lines of mice by inserting EYFP or ECFP cDNAs into the ROSA26 locus, preceded by a loxP-flanked stop sequence. These strains were tested by crossing them with transgenic strains expressing Cre in a ubiquitous (β-actin-Cre) or a cell-specific (Isl1-Cre and En1-Cre) pattern. The resulting EYFP or ECFP expression patterns indicated that the reporter strains function as faithful monitors of Cre activity.ConclusionsIn contrast to existing lacZ reporter lines, where lacZ expression cannot easily be detected in living tissue, the EYFP and ECFP reporter strains are useful for monitoring the expression of Cre and tracing the lineage of these cells and their descendants in cultured embryos or organs. The non-overlapping emission spectra of EYFP and ECFP make them ideal for double labeling studies in living tissues.

Subfields: Biophysics, Genetics, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/bbca7a6ffa5b4d1a8161a43569192899c33c21a5

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Experience dictates stem cell fate in the adult hippocampus. (2011, positive): “Nestin-CreERT2 animals were bred with R26R enhanced yellow fluorescent protein (EYFP) reporter mice (Srinivas et al., 2001).”
The notch ligands Dll4 and Jagged1 have opposing effects on angiogenesis. (2009, positive): “For mosaic deletion of Jagged1 in the retina, Jag1floxed/floxed were combined with VE-Cadherin(PAC)-CreERT2 (R.H.A., unpublished data) and Rosa26 EYFP Cre reporter (Srinivas et al., 2001) transgenes.”
Isl1 identifies a cardiac progenitor population that proliferates prior to differentiation and contributes a majority of cells to the heart. (2003, methodology): “Jessell and have been previously described (Srinivas et al., 2001).”
Sox1 marks an activated neural stem/progenitor cell in the hippocampus (2012, background): “LC-1, R26R, R26eYFP and Z/EG mice were previously published (Soriano, 1999; Novak et al., 2000; Srinivas et al., 2001; Schönig et al., 2002).”
Macrophage-Induced Blood Vessels Guide Schwann Cell-Mediated Regeneration of Peripheral Nerves (2015, background): “The mice also contained a floxed YFPreporter gene in the Rosa26 locus to monitor the efficiency of CRE-mediated recombination (Srinivas et al., 2001).”
Fate tracing reveals the endothelial origin of hematopoietic stem cells (2008, methodology): “VE-cadherin Cre and inducible VE-cadherin Cre mice, described elsewhere (Alva et al., 2006; Monvoisin et al., 2006), were crossed to R26R Cre reporter lines with lacZ reporter (Soriano, 1999) and separately EYFP reporter (Srinivas et al., 2001).”
Acquisition of granule neuron precursor identity is a critical determinant of progenitor cell competence to form Hedgehog-induced medulloblastoma (2008, positive): “To establish a possible lineage relationship between hGFAPcre and Olig2+ cells, we fate mapped hGFAP-cre-expressing precursors with a conditional ROSA26-eYPFflox/flox reporter (Srinivas et al., 2001).”
EphB signaling controls lineage plasticity of adult neural stem cell niche cells (2010, neutral): “, 2007), R26R-LacZ (Soriano, 1999), and R26R-YFP (Srinivas et al., 2001) mice on”
Multiple layers of B cell memory with different effector functions (2009, background): “We further crossed that transgenic mouse with the Rosa26-loxP-EYFP reporter mous”