Nature Genetics
36, 283 - 287 (2004)
Published online: 22 February 2004; | doi:10.1038/ng1314
A complementary transposon tool kit for Drosophila melanogaster using P and piggyBacStephen T Thibault1, Matthew A Singer1, 2, Wesley Y Miyazaki1, Brett Milash1, 2, Nicholas A Dompe1, Carol M Singh1, 2, Ross Buchholz1, Madelyn Demsky1, Robert Fawcett1, Helen L Francis-Lang1, Lisa Ryner1, Lai Man Cheung1, Angela Chong1, Cathy Erickson1, William W Fisher1, Kimberly Greer1, Stephanie R Hartouni1, Elizabeth Howie1, Lakshmi Jakkula1, Daniel Joo1, 2, Keith Killpack1, Alex Laufer1, 2, Julie Mazzotta1, Ronald D Smith1, Lynn M Stevens1, Christiana Stuber1, Lory R Tan1, Richard Ventura1, Alesa Woo1, Irena Zakrajsek1, 2, Lora Zhao1, Feng Chen1, Candace Swimmer1, Casey Kopczynski1, 2, Geoffrey Duyk1, Margaret L Winberg1
& Jonathan Margolis11
Exelixis, 170 Harbor Way, South San Francisco, California 94083-0511, USA. 2
Present addresses: Chemicon International, 28820 Single Oak Drive, Temecula, California 92590, USA (M.A.S.); Huntsman Cancer Institute, 2000 Circle of Hope, University of Utah, Salt Lake City, Utah 84132, USA (B.M.); EnVivo Pharmaceuticals, 480 Arsenal Street, Building 1, Watertown, Massachusetts 02472, USA (C.M.S.); Metabolex, 3876 Bay Center Place, Hayward, California 94545, USA (D.J.); Renovis, Two Corporate Dr., South San Francisco, California 94080, USA (A.L.); Genentech, 1 DNA Way, South San Francisco, California 94080, USA (I.Z.); Ercole Biotech, 7030 Kit Creek Road, P.O. Box 12295, Research Triangle Park, North Carolina 27709, USA (C.K.).
Correspondence should be addressed to Stephen T Thibault thibault@exelixis.comWith the availability of complete genome sequence for Drosophila melanogaster, one of the next strategic goals for fly researchers is a complete gene knockout collection. The P-element transposon1, the workhorse of D. melanogaster molecular genetics, has a pronounced nonrandom insertion spectrum2. It has been estimated that 87% saturation of the  13,500-gene complement of D. melanogaster
3 might require generating and analyzing up to 150,000 insertions2. We describe specific improvements to the lepidopteran transposon piggyBac
4 and the P element that enabled us to tag and disrupt genes in D. melanogaster more efficiently. We generated over 29,000 inserts resulting in 53% gene saturation and a more diverse collection of phenotypically stronger insertional alleles. We found that piggyBac has distinct global and local gene-tagging behavior from that of P elements. Notably, piggyBac excisions from the germ line are nearly always precise, piggyBac does not share chromosomal hotspots associated with P and piggyBac is more effective at gene disruption because it lacks the P bias for insertion in 5' regulatory sequences.
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