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Related Concept Videos

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Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines
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Intragenic Enhancers Attenuate Host Gene Expression.

Senthilkumar Cinghu1, Pengyi Yang1, Justin P Kosak1

  • 1Epigenetics & Stem Cell Biology Laboratory, National Institute of Environmental Health Sciences, NIH, Research Triangle Park, NC 27709, USA.

Molecular Cell
|October 7, 2017
PubMed
Summary

Intragenic enhancers regulate gene transcription by interfering with productive elongation. This transcription interference, not enhancer RNAs, attenuates host gene expression and influences cell fate determination.

Keywords:
enhancerenhancer RNAenhancer transcriptiongeneticsgenomicstranscription regulation

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Eukaryotic gene transcription involves multiple regulatory steps, but mechanisms governing productive elongation are unclear.
  • Enhancers activate gene transcription via RNA polymerase II (Pol II) but their transcription and enhancer RNAs (eRNAs) roles are not fully understood.

Purpose of the Study:

  • To investigate the role of transcription at intragenic enhancers during productive elongation.
  • To elucidate the mechanisms by which enhancers influence host gene transcription and cell fate.

Main Methods:

  • CRISPR/Cas9-mediated gene editing to delete intragenic enhancers.
  • Analysis of nascent eRNA expression and host gene transcription.
  • Assessment of cell fate determination.

Main Results:

  • Transcription at intragenic enhancers interferes with and attenuates host gene transcription during productive elongation.
  • Attenuation correlates with nascent eRNA levels, but is caused by the transcription process itself, not the eRNAs.
  • CRISPR/Cas9 deletions confirmed a physiological role for this attenuation in cell fate determination.

Conclusions:

  • Intragenic enhancers fine-tune host gene transcription via transcription interference during productive elongation.
  • This mechanism allows regulatory elements to serve dual functions: enhancing transcription from a distance and modulating host gene expression.
  • Intragenic enhancer-mediated transcription attenuation plays a critical role in cell fate determination.