TALE-mediated modulation of transcriptional enhancers in vivo

• Published in: Nature methods Vol. 10; no. 8; pp. 762 - 767
• Main Authors: Crocker, Justin, Stern, David L
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.08.2013; Nature Publishing Group
• Subjects: 631/1647/1511; 631/1647/1513/1967/2316; 631/208/200; 631/337/572; Analysis; Animals; Bioassays; Bioinformatics; Biological Microscopy; Biological Techniques; Biomedical Engineering/Biotechnology; Deoxyribonucleic acid; DNA; Drosophila; Drosophila melanogaster - genetics; Drosophila Proteins - genetics; Enhancer Elements, Genetic; Female; Gene expression; Gene Expression Regulation, Developmental; Genetic aspects; Genetic transcription; Genome, Insect; Genomes; Genomics; Homeodomain Proteins - genetics; Insects; Larvae; Life Sciences; Male; Microscopy, Confocal; Organisms, Genetically Modified; Proteins; Proteomics; Transcription Factors - genetics; Transcriptional Activation
• ISSN: 1548-7091; 1548-7105
• DOI: 10.1038/nmeth.2543

Transcription activator–like effectors are used for in vivo activation and repression of endogenous promoters and enhancers in the fruit fly. We tested whether transcription activator–like effectors (TALEs) could mediate repression and activation of endogenous enhancers in the Drosophila genome. TALE repressors (TALERs) targeting each of the five even-skipped ( eve ) stripe enhancers generated repression specifically of the focal stripes. TALE activators (TALEAs) targeting the eve promoter or enhancers caused increased expression primarily in cells normally activated by the promoter or targeted enhancer, respectively. This effect supports the view that repression acts in a dominant fashion on transcriptional activators and that the activity state of an enhancer influences TALE binding or the ability of the VP16 domain to enhance transcription. In these assays, the Hairy repression domain did not exhibit previously described long-range transcriptional repression activity. The phenotypic effects of TALER and TALEA expression in larvae and adults are consistent with the observed modulations of eve expression. TALEs thus provide a novel tool for detection and functional modulation of transcriptional enhancers in their native genomic context.