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Nat Commun ; 10(1): 2262, 2019 05 22.
Artigo em Inglês | MEDLINE | ID: mdl-31118412

RESUMO

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.


Assuntos
Edição de Genes/métodos , Vetores Genéticos/genética , Integrases/genética , Plasmídeos/genética , Animais , Técnicas de Cultura de Células , Clonagem Molecular/métodos , Camundongos , Camundongos Transgênicos , Células-Tronco Embrionárias Murinas , Regiões Promotoras Genéticas/efeitos dos fármacos , Regiões Promotoras Genéticas/genética , Recombinação Genética/efeitos dos fármacos , Tamoxifeno/farmacologia
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