Red Light/Green Light, a Dual Fluorescent Protein Reporter System To Study Enhancer-Promoter Specificity in Drosophila

Enhancers activate gene transcription in spatial and temporal patterns by interactions with gene promoters. These elements typically reside distal to their target promoter, with which they must interact selectively. Additional elements may contribute to enhancer-promoter specificity, including remot...

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Bibliographic Details
Published in:G3 : genes - genomes - genetics 2020-03, Vol.10 (3), p.985-997
Main Authors: Camino, Eric M, Weinstein, Micheal L, List, Mary P, Vellky, Jordan E, Rebeiz, Mark, Williams, Thomas M
Format: Article
Language:English
Subjects:
Animals
Animals, Genetically Modified
drosophila
Drosophila melanogaster - genetics
enhancer
Enhancer Elements, Genetic
Female
Fluorescence
Genes, Reporter
Insect Proteins - genetics
Investigations
Male
promoter
Promoter Regions, Genetic
remote control element
tethering element
Transgenes
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Summary:Enhancers activate gene transcription in spatial and temporal patterns by interactions with gene promoters. These elements typically reside distal to their target promoter, with which they must interact selectively. Additional elements may contribute to enhancer-promoter specificity, including remote control element sequences within enhancers, tethering elements near promoters, and insulator/boundary elements that disrupt off-target interactions. However, few of these elements have been mapped, and as a result, the mechanisms by which these elements interact remain poorly understood. One impediment is their method of study, namely reporter transgenes in which enhancers are placed adjacent to a heterologous promoter, which may circumvent mechanisms controlling enhancer-promoter specificity and long-range interactions. Here, we report an optimized dual reporter transgene system in that allows the simultaneous comparison of an enhancer's ability to activate proximal and distal fluorescent reporter genes. Testing a panel of fluorescent transgenes , we found a two-protein combination that allows simultaneous measurement with minimal detection interference. We note differences among four tested enhancers in their ability to regulate a distally placed reporter transgene. These results suggest that enhancers differ in their requirements for promoter interaction and raise important practical considerations when studying enhancer function.
ISSN:2160-1836
2160-1836
DOI:10.1534/g3.119.401033