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Nat Genet. 2004 Mar;36(3):283-7. Epub 2004 Feb 22.

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

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1
Exelixis, 170 Harbor Way, South San Francisco, California 94083-0511, USA. thibault@exelixis.com

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, 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.

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PMID:
14981521
DOI:
10.1038/ng1314
[Indexed for MEDLINE]
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