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Nat Commun ; 10(1): 2262, 2019 05 22.
Article in English | MEDLINE | ID: mdl-31118412

ABSTRACT

Most biomedical research aimed at understanding gene function uses the Cre-Lox system, which consists of the Cre recombinase-dependent deletion of genes containing LoxP sites. This system enables conditional genetic modifications because the expression and activity of the recombinase Cre/CreERT2 can be regulated in space by tissue-specific promoters and in time by the ligand tamoxifen. Since the precise Cre-Lox recombination event is invisible, methods were developed to report Cre activity and are widely used. However, numerous studies have shown that expression of a given Cre activity reporter cannot be assumed to indicate deletion of other LoxP-flanked genes of interest. Here, we report the generation of an inducible dual reporter-Cre mouse allele, iSuRe-Cre. By significantly increasing Cre activity in reporter-expressing cells, iSuRe-Cre provides certainty that these cells have completely recombined floxed alleles. This genetic tool increases the ease, efficiency, and reliability of conditional mutagenesis and gene function analysis.


Subject(s)
Gene Editing/methods , Genetic Vectors/genetics , Integrases/genetics , Plasmids/genetics , Animals , Cell Culture Techniques , Cloning, Molecular/methods , Mice , Mice, Transgenic , Mouse Embryonic Stem Cells , Promoter Regions, Genetic/drug effects , Promoter Regions, Genetic/genetics , Recombination, Genetic/drug effects , Tamoxifen/pharmacology
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