A complementary transposon tool kit for Drosophila melanogaster using P and piggyBac

Autor: Ross Buchholz, Jonathan Margolis, Kimberly Greer, Carol M. Singh, Lory R Tan, Lynn M Stevens, Cathy Erickson, Julie Mazzotta, Wesley Y Miyazaki, Lora Zhao, Angela Chong, Robert Fawcett, William W Fisher, Lai Man Cheung, Keith Killpack, Alex Laufer, Geoffrey Duyk, Stephen T Thibault, Candace Swimmer, Matthew A Singer, Helen Francis-Lang, Casey Kopczynski, Elizabeth Howie, Lakshmi Jakkula, Madelyn Robin Demsky, Brett Milash, Irena Zakrajsek, Richard Benn Abegania Ventura, Feng Chen, Nicholas A Dompe, Margaret L. Winberg, Alesa Woo, Lisa Ryner, Daniel Joo, Ronald D. Smith, Stephanie R Hartouni, Christiana Stuber
Rok vydání: 2004
Předmět:
Zdroj: Nature Genetics. 36:283-287
ISSN: 1546-1718
1061-4036
DOI: 10.1038/ng1314
Popis: With 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 transposon, the workhorse of D. melanogaster molecular genetics, has a pronounced nonrandom insertion spectrum. It has been estimated that 87% saturation of the approximately 13,500-gene complement of D. melanogaster might require generating and analyzing up to 150,000 insertions. We describe specific improvements to the lepidopteran transposon piggyBac 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.
Databáze: OpenAIRE
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