Abstract
The Cre/loxP site-specific recombination system has been used successfully for genome manipulation in a wide range of species. However, in Drosophila melanogaster, a major model organism for genetic analyses, the alternative FLP/FRT system, which is less efficient at least in mammalian cells, has been established, primarily for the generation of genetic mosaics for clonal analyses. To extend genetic methodology in D. melanogaster, we have created transgenic lines allowing tissue-specific expression of Cre recombinase with the UAS/GAL4 system. Surprisingly, chronic expression of Cre recombinase from these transgenes (UAST-cre) was found to be toxic for proliferating cells. Therefore, we also generated transgenic lines allowing the expression of Cre recombinase fused to the ligand-binding domain of the human estrogen receptor (UASP-cre-EBD). We demonstrate that recombination can be efficiently dissociated from toxicity by estrogen-dependent regulation of recombinase activity of the UASP-cre-EBD transgene products.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Animals, Genetically Modified
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Apoptosis / drug effects
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Attachment Sites, Microbiological / genetics
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Cell Division / drug effects
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Cell Line
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Dose-Response Relationship, Drug
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Drosophila melanogaster / cytology*
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Drosophila melanogaster / drug effects*
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Drosophila melanogaster / genetics
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Enzyme Activation / drug effects
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Estradiol / pharmacology*
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Eye / growth & development
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Eye / metabolism
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Eye / ultrastructure
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Gene Expression Regulation, Developmental / drug effects*
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Humans
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Integrases / genetics
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Integrases / metabolism*
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Integrases / toxicity
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Mutagenesis, Site-Directed / drug effects
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Mutagenesis, Site-Directed / genetics
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Organ Specificity
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Receptors, Estrogen / genetics
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Receptors, Estrogen / metabolism
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Recombinant Fusion Proteins / toxicity
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Recombination, Genetic / drug effects
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Recombination, Genetic / genetics
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Transgenes / genetics
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Viral Proteins / genetics
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Viral Proteins / metabolism*
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Viral Proteins / toxicity
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Wings, Animal / growth & development
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Wings, Animal / metabolism
Substances
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Receptors, Estrogen
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Recombinant Fusion Proteins
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Viral Proteins
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Estradiol
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Cre recombinase
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Integrases