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

A complementary transposon tool kit for Drosophila melanogaster using P and piggyBac 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 1 , the workhorse of D. melanogaster molecular genetics, has a pronounced nonrandom insertion spectrum 2 . 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 insertions 2 . 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Nature Genetics Springer Journals

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

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References (34)

Publisher
Springer Journals
Copyright
Copyright © 2004 by Nature Publishing Group
Subject
Biomedicine; Biomedicine, general; Human Genetics; Cancer Research; Agriculture; Gene Function; Animal Genetics and Genomics
ISSN
1061-4036
eISSN
1546-1718
DOI
10.1038/ng1314
Publisher site
See Article on Publisher Site

Abstract

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 1 , the workhorse of D. melanogaster molecular genetics, has a pronounced nonrandom insertion spectrum 2 . 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 insertions 2 . 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.

Journal

Nature Genetics – Springer Journals

Published: Feb 22, 2004

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