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

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ID: 266622
2001
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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.
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srinivas2001bmccre Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Shankar Srinivas;Tomoko Watanabe;Chyuan-Sheng Lin;Chris M William;Yasuto Tanabe;Thomas M Jessell;Frank Costantini;Shankar Srinivas;Tomoko Watanabe;Chyuan-Sheng Lin;Chris M William;Yasuto Tanabe;Thomas M Jessell;Frank Costantini;
Journal bmc developmental biology
Year 2001
DOI
doi:10.1186/1471-213X-1-4
URL
http://dx.doi.org/10.1186/1471-213X-1-4
Keywords
developmental biology life sciences animal models general National Center for Biotechnology Information NCBI NLM MEDLINE Mice Inbred C57BL RNA animals pubmed abstract nih national institutes of health national library of medicine research support non-u.s. gov't u.s. gov't female P.H.S. cell line Genes recombination bacterial proteins / genetics* stem cells / metabolism brain / metabolism genetic vectors green fluorescent proteins proteins / genetics* embryo transgenic transfection viral proteins / genetics* genetic markers / genetics* mutagenesis inbred strains mammalian genetic / genetics luminescent proteins / biosynthesis integrases / genetics* pmid:11299042 pmc31338 doi:10.1186/1471-213x-1-4 s srinivas t watanabe f costantini bacterial proteins / biosynthesis brain / cytology gene targeting / methods* reporter / genetics* luminescent proteins / genetics* insertional untranslated stem cells / chemistry

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