Reference - Detail
| 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 |
| Altmetric score |
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 18 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Drosophila | DGRC#115635 |