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TALE-mediated modulation of transcriptional enhancers in vivo

Nature Methods volume 10, pages 762–767 (2013)Cite this article

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Abstract

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.

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Figure 1: TALERs targeted to the promoter can repress expression of eve.
Figure 2: TALE targeted activation of the eve promoter.
Figure 3: TALER targeted repression of the eve stripe 2 enhancer.
Figure 4: TALE targeted repression and activation of eve stripe enhancers.
Figure 5: Phenotypes resulting from enhancer-TALEA-VP64 activation of eve enhancers.

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Acknowledgements

We thank T. Martin and A. DePace (Harvard Medical School) for the eveS2-lacZ enhancer flies; E. Preger-Ben Noon and C. Standley for comments on the manuscript; and B. Pfeiffer, M. Schroeder, R. Mann and the entire Stern lab for discussion. We also thank the Janelia Farm Research Campus community for facilitating this work and for providing an inspiring scientific environment.

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  1. Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia, USA

    Justin Crocker & David L Stern

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  1. Justin Crocker
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  2. David L Stern
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J.C. conceived of, designed and executed the experiments and analyzed the data, with mentorship from D.L.S. J.C. and D.L.S. wrote the manuscript.

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Correspondence to Justin Crocker.

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The authors declare no competing financial interests.

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Crocker, J., Stern, D. TALE-mediated modulation of transcriptional enhancers in vivo. Nat Methods 10, 762–767 (2013). https://doi.org/10.1038/nmeth.2543

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