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Open Access Highly Accessed Methodology article

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

Shankar Srinivas14, Tomoko Watanabe1, Chyuan-Sheng Lin2, Chris M William3, Yasuto Tanabe3, Thomas M Jessell3 and Frank Costantini1*

Author Affiliations

1 Department of Genetics and Development, Columbia University, New York, USA

2 Herbert Irving Comprehensive Cancer Center, Columbia University, New York, USA

3 Howard Hughes Medical Institute, Department of Biochemistry and Molecular Biophysics, and Center for Neurobiology and Behavior, Columbia University, New York, USA

4 Present address: National Institute for Medical Research, The Ridgeway, Mill Hill, London, United Kingdom

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BMC Developmental Biology 2001, 1:4  doi:10.1186/1471-213X-1-4

Published: 27 March 2001

Abstract

Background

Several 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.

Results

In 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.

Conclusions

In 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.