| RRC ID |
30452
|
| Author |
Horn C, Offen N, Nystedt S, Häcker U, Wimmer EA.
|
| Title |
piggyBac-based insertional mutagenesis and enhancer detection as a tool for functional insect genomics.
|
| Journal |
Genetics
|
| Abstract |
Transposon mutagenesis provides a fundamental tool for functional genomics. Here we present a non-species-specific, combined enhancer detection and binary expression system based on the transposable element piggyBac: For the different components of this insertional mutagenesis system, we used widely applicable transposons and distinguishable broad-range transformation markers, which should enable this system to be operational in nonmodel arthropods. In a pilot screen in Drosophila melanogaster, piggyBac mutator elements on the X chromosome were mobilized in males by a Hermes-based jumpstarter element providing piggyBac transposase activity under control of the alpha1-tubulin promoter. As primary reporters in the piggyBac mutator elements, we employed the heterologous transactivators GAL4delta or tTA. To identify larval and adult enhancer detectors, strains carrying UASp-EYFP or TRE-EYFP as secondary reporter elements were used. Tissue-specific enhancer activities were readily observed in the GAL4delta/UASp-based systems, but only rarely in the tTA/TRE system. Novel autosomal insertions were recovered with an average jumping rate of 80%. Of these novel insertions, 3.8% showed homozygous lethality, which was reversible by piggyBac excision. Insertions were found in both coding and noncoding regions of characterized genes and also in noncharacterized and non-P-targeted CG-number genes. This indicates that piggyBac will greatly facilitate the intended saturation mutagenesis in Drosophila.
|
| Volume |
163(2)
|
| Pages |
647-61
|
| Published |
2003-2-1
|
| DOI |
10.1093/genetics/163.2.647
|
| PMID |
12618403
|
| PMC |
PMC1462455
|
| MeSH |
Animals
DNA Transposable Elements*
Drosophila melanogaster / genetics*
Enhancer Elements, Genetic*
Genes, Reporter
Genomics*
Mutagenesis, Insertional
Mutation
|
| IF |
4.015
|
| Times Cited |
113
|
|
WOS Category
|
GENETICS & HEREDITY
|
| Resource |
| Drosophila |
DGRC#115635 |
