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Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines

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Evolved Repression Overcomes Enhancer Robustness.

Ella Preger-Ben Noon¹, Fred P Davis¹, David L Stern¹

¹Janelia Research Campus, Howard Hughes Medical Institute, 19700 Helix Drive, Ashburn, VA 20147, USA.

Developmental Cell

|November 15, 2016

Summary

This summary is machine-generated.

Robust enhancers can evolve loss of expression by recruiting repressor binding sites. This mechanism, observed in the shavenbaby gene, can overcome robustness and minimize pleiotropic effects during evolution.

Keywords:

enhancer evolution morphology repression robustness transcription

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Area of Science:

Developmental biology
Evolutionary genetics
Molecular biology

Background:

Biological systems exhibit remarkable robustness, particularly transcriptional enhancers.
Enhancer robustness often arises from clustered transcription factor binding sites.
Mechanisms by which robust enhancers evolve loss of function via point mutations remain unclear.

Purpose of the Study:

To functionally dissect a robust transcriptional enhancer of the shavenbaby gene.
To understand how enhancer robustness can be overcome during evolution.
To investigate the role of repressor binding sites in evolutionary changes of enhancer activity.

Main Methods:

High-resolution functional dissection of the shavenbaby enhancer.
Analysis of transcription factor binding sites, including those for Arrowhead and Abrupt.
Comparative analysis of enhancer activity during evolution.

Main Results:

Enhancer robustness is encoded by multiple binding sites for the transcriptional activator Arrowhead.
Evolutionary loss of Arrowhead sites weakened enhancer activity.
Complete silencing of enhancer function required the evolution of a binding site for the repressor Abrupt.

Conclusions:

Recruitment of repressor binding sites can overcome enhancer robustness.
Repressor recruitment may be a general mechanism for breaking robust regulatory linkages during evolution.
This process can minimize pleiotropic consequences of enhancer evolution.