Ligand-regulated site-specific recombination.
Site-specific recombination offers a potential way to alter a living genome by design in a precise and stable manner. This potential requires strategies which can be used to regulate the recombination event. We describe a strategy to regulate FLP recombinase activity which relies on expressing FLP as a fusion protein with steroid hormone receptor ligand binding domains (LBDs). In the absence of a ligand cognate to the LBD, the recombinase activity of the fusion protein is extremely low. Upon ligand administration, recombinase activity is rapidly induced. These results outline the basis for inducible expression or disruption strategies based on inducible recombination. Additionally, we have exploited the conditional nature of FLP-LBD fusion proteins to direct integration of a plasmid into a specific genomic site at frequencies approaching the frequency of random integration.
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Gene Expression Program, European Molecular Biology Laboratory, Heidelberg, Federal Republic of Germany.
Stewart
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D000818:Animals; D002460:Cell Line; D004254:DNA Nucleotidyltransferases / Q000096:biosynthesis / Q000378:metabolism*; D004958:Estradiol / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008024:Ligands; D011944:Receptors, Androgen / Q000096:biosynthesis / Q000378:metabolism; D011960:Receptors, Estrogen / Q000096:biosynthesis / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000096:biosynthesis / Q000378:metabolism; D011987:Receptors, Steroid / Q000096:biosynthesis / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism; D011995:Recombination, Genetic; D015183:Restriction Mapping; D014162:Transfection
D008024:Ligands; D011944:Receptors, Androgen; D011960:Receptors, Estrogen; D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D011993:Recombinant Fusion Proteins; D004958:Estradiol; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase