We are analyzing https://link.springer.com/article/10.1186/1471-213x-1-4.

Title:
Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus | BMC Developmental Biology
Description:
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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Keywords {🔍}

mice, expression, article, pubmed, reporter, cre, eyfp, cells, ecfp, google, scholar, cas, rosa, sequence, plasmid, strains, lacz, targeting, digested, resulting, site, locus, targeted, strain, inserted, gene, cremediated, genomic, figure, paci, cell, islcre, embryos, allele, fluorescent, protein, transgenic, embryo, asci, kpni, excision, lineage, vector, original, mouse, data, stop, green, recombination, express,

Topics {✒️}

β-actin promoter/cmv enhancer multiple cloning site adenovirus splice acceptor recombinase-mediated gene activation green fluorescent protein β-actin-cre transgene cre-mediated chromosome loss chyuan-sheng lin cyan fluorescent protein mutate developmental genes cre-mediated excision event motor neuron-dependent step p1 site-specific recombination β-actin/cre mice related subjects cre-expressing transgenic mice characterized r26r-lacz strains genetic ablation reveals frank costantini embryonic stem cells entire floxed neo-tpa floxed neo-tpa cassette tissue-specific cre strains conditional gene targeting pgk-neo selectable marker doubly transgenic offspring loxp-neo-tpa-loxp r26r-lacz reporter mice conditional knockout alleles r26r-yfp targeting construct loxp-flanked stop sequence article srinivas reporter mouse strain sv40 polyadenylation sequence r26r-ecfp colonies carried β-actin-cre authors’ original file privacy choices/manage cookies medical research heterozygous isl1-cre mice express enhanced yellow lim homeobox genes cre reporter strains cre-mediated recombination reporter proteins diphtheria toxin gene cre-loxp system rosa26 genomic arms improved reporter strain cre-mediated excision

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         headline:Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus
         description: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. 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. 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.
         datePublished:2001-03-27T00:00:00Z
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            Splice Acceptor
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      headline:Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus
      description: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. 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. 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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         Green Fluorescent Protein
         PacI
         ApaI
         Multiple Cloning Site
         Splice Acceptor
         Developmental Biology
         Animal Models
         Life Sciences
         general
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      name:Department of Genetics and Development, Columbia University, New York, USA

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