Genetics, Vol 144, 715-726, Copyright © 1996

INVESTIGATIONS

Transgene Coplacement and High Efficiency Site-Specific Recombination With the Cre/loxP System in Drosophila

M. L. Siegal and D. L. Hartl
Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts 02138

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 F(1) progeny. Excision in germ cells of the F(1) yields a strong grand-maternal effect, observed as a highly skewed ratio of eye-color phenotypes in the F(2) 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.

This article has been cited by other articles:

				D. Gohl, M. Muller, V. Pirrotta, M. Affolter, and P. Schedl Enhancer Blocking and Transvection at the Drosophila apterous Locus Genetics, January 1, 2008; 178(1): 127 - 143. [Abstract] [Full Text] [PDF]

				K. J. T. Venken and H. J. Bellen Transgenesis upgrades for Drosophila melanogaster Development, October 15, 2007; 134(20): 3571 - 3584. [Abstract] [Full Text] [PDF]

				J. Bischof, R. K. Maeda, M. Hediger, F. Karch, and K. Basler An optimized transgenesis system for Drosophila using germ-line-specific {varphi}C31 integrases PNAS, February 27, 2007; 104(9): 3312 - 3317. [Abstract] [Full Text] [PDF]

				J. Mihaly, S. Barges, L. Sipos, R. Maeda, F. Cleard, I. Hogga, W. Bender, H. Gyurkovics, and F. Karch Dissecting the regulatory landscape of the Abd-B gene of the bithorax complex Development, August 1, 2006; 133(15): 2983 - 2993. [Abstract] [Full Text] [PDF]

				J. R. Bateman, A. M. Lee, and C.-t. Wu Site-Specific Transformation of Drosophila via {phi}C31 Integrase-Mediated Cassette Exchange Genetics, June 1, 2006; 173(2): 769 - 777. [Abstract] [Full Text] [PDF]

				W. J. Gong and K. G. Golic Loss of Hsp70 in Drosophila Is Pleiotropic, With Effects on Thermotolerance, Recovery From Heat Shock and Neurodegeneration Genetics, January 1, 2006; 172(1): 275 - 286. [Abstract] [Full Text] [PDF]

				T. Shimizu, T. Oishi, A. Omori, A. Sugiura, K. Hirota, H. Aoyama, T. Saito, T. Sugaya, Y. Kon, J. D. Engel, et al. Identification of cis-Regulatory Sequences in the Human Angiotensinogen Gene by Transgene Coplacement and Site-Specific Recombination Mol. Cell. Biol., April 15, 2005; 25(8): 2938 - 2945. [Abstract] [Full Text] [PDF]

				S. Schmitt, M. Prestel, and R. Paro Intergenic transcription through a Polycomb group response element counteracts silencing Genes & Dev., March 15, 2005; 19(6): 697 - 708. [Abstract] [Full Text] [PDF]

				W. J. Gong and K. G. Golic Genomic Deletions of the Drosophila melanogaster Hsp70 Genes Genetics, November 1, 2004; 168(3): 1467 - 1476. [Abstract] [Full Text] [PDF]

				H. B. Xie and K. G. Golic Gene Deletions by Ends-In Targeting in Drosophila melanogaster Genetics, November 1, 2004; 168(3): 1477 - 1489. [Abstract] [Full Text] [PDF]

				J. Parsch Functional Analysis of Drosophila melanogaster Gene Regulatory Sequences by Transgene Coplacement Genetics, September 1, 2004; 168(1): 559 - 561. [Abstract] [Full Text] [PDF]

				D. Egli, E. Hafen, and W. Schaffner An Efficient Method to Generate Chromosomal Rearrangements by Targeted DNA Double-Strand Breaks in Drosophila melanogaster Genome Res., July 1, 2004; 14(7): 1382 - 1393. [Abstract] [Full Text] [PDF]

				A. C. Groth, M. Fish, R. Nusse, and M. P. Calos Construction of Transgenic Drosophila by Using the Site-Specific Integrase From Phage {phi}C31 Genetics, April 1, 2004; 166(4): 1775 - 1782. [Abstract] [Full Text] [PDF]

				E. J. Kuhn, C. M. Hart, and P. K. Geyer Studies of the Role of the Drosophila scs and scs' Insulators in Defining Boundaries of a Chromosome Puff Mol. Cell. Biol., February 15, 2004; 24(4): 1470 - 1480. [Abstract] [Full Text] [PDF]

				N. Jasinskiene, C. J. Coates, A. Ashikyan, and A. A. James High efficiency, site-specific excision of a marker gene by the phage P1 cre-loxP system in the yellow fever mosquito, Aedes aegypti Nucleic Acids Res., November 15, 2003; 31(22): e147 - e147. [Abstract] [Full Text] [PDF]

				T. J. Parnell, M. M. Viering, A. Skjesol, C. Helou, E. J. Kuhn, and P. K. Geyer An endogenous Suppressor of Hairy-wing insulator separates regulatory domains in Drosophila PNAS, November 11, 2003; 100(23): 13436 - 13441. [Abstract] [Full Text] [PDF]

				W. J. Gong and K. G. Golic Ends-out, or replacement, gene targeting in Drosophila PNAS, March 4, 2003; 100(5): 2556 - 2561. [Abstract] [Full Text] [PDF]

				J.-L. Chen, K. L. Huisinga, M. M. Viering, S. A. Ou, C.-t. Wu, and P. K. Geyer Enhancer action in trans is permitted throughout the Drosophila genome PNAS, March 19, 2002; 99(6): 3723 - 3728. [Abstract] [Full Text] [PDF]

				J. E.F. Butler and J. T. Kadonaga Enhancer-promoter specificity mediated by DPE or TATA core promoter motifs Genes & Dev., October 1, 2001; 15(19): 2515 - 2519. [Abstract] [Full Text] [PDF]

				M. L. Siegal and D. L. Hartl An experimental test for lineage-specific position effects on alcohol dehydrogenase (Adh) genes in Drosophila PNAS, December 22, 1998; 95(26): 15513 - 15518. [Abstract] [Full Text] [PDF]