genesis · 2010 · 31 citations · 19 references
Life SciencesGeneticsMolecular GeneticsGene ExpressionGene RegulationTranscriptional RegulationCis-regulatory ElementsGene Expression RegulationGene Regulatory ElementsGenome Integration StrategiesRegulatory Element EngineeringRegulatory GenomicsCis-acting Regulatory ElementsDrosophila ModelsEnhancer ElementsFluorescent LabelingHigh-throughput Reporter AssaysReporter Gene ConstructionSite-specific IntegrationTranscriptional Control Elements
The fruit fly Drosophila is a leading model system for the study of transcriptional control by cis-regulatory elements or enhancers. Here, we present a rapid and highly efficient system for the large-scale analysis of enhancer elements, site-specifically integrated into the Drosophila genome. This system, which is scalable for either small projects or high-throughput approaches, makes use of the Gateway cloning technology and the PhiC31 site-specific integration system, which allows the insertion of constructs at predetermined genomic locations. Thus, this system allows not only a fast and easy analysis of reporter gene expression in live animals, but also the simultaneous analysis of different regulatory outputs on a cellular resolution by recombining in the same animal distinct enhancer elements fused to different fluorescent proteins.
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