Transgene Coplacement and high efficiency site-specific recombination with the Cre/loxP system in Drosophila.
Studies of gene function and regulation in transgenic Drosophila are often compromised by the possibility of genomic position effects on gene expression. We have developed a method called transgene coplacement, in which any two sequences can be positioned at exactly the same site and orientation in the genome. Transgene coplacement makes use of the bacteriophage P1 system of Cre/loxP site-specific recombination, which we have introduced into Drosophila. In the presence of a cre transgene driven by a dual hsp70-Mos1 promoter, a white reporter gene flanked by loxP sites is excised with virtually 100% efficiency both in somatic cells and in germ cells. A strong maternal effect, resulting from Cre recombinase present in the oocyte, is observed as white or mosaic eye color in F1 progeny. Excision in germ cells of the F1 yields a strong grand-maternal effect, observed as a highly skewed ratio of eye-color phenotypes in the F2 generation. The excision reactions of Cre/loxP and the related FLP/FRT system are used to create Drosophila lines in which transgenes are at exactly allelic sites in homologous chromosomes.
Siegal
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ML
Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
Hartl
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DL
D018528:ATP-Binding Cassette Transporters; D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D004247:DNA; D029721:Drosophila Proteins; D005136:Eye Proteins; D005260:Female; D015870:Gene Expression; D017930:Genes, Reporter; D005854:Germ Cells; D007301:Insect Hormones / Q000235:genetics*; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D011995:Recombination, Genetic; D019076:Transgenes; D014764:Viral Proteins
D018528:ATP-Binding Cassette Transporters; D029721:Drosophila Proteins; D005136:Eye Proteins; D007301:Insect Hormones; D014764:Viral Proteins; C059254:w protein, Drosophila; D004247:DNA; C045073:Cre recombinase; D019426:Integrases