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The cells of the blastocyst inner cell mass only remain pluripotent for a short time. This state of pluripotency and self-renewal can be maintained in embryonic stem (ES) cell culture by adding specific chemicals or growth factors to ensure the cells can continue dividing and later differentiate into different cell types. In some cases, the cells are grown on a feeder layer of differentiated cells, which provides the growth factors and extracellular matrix components necessary for stem cell...
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Transcription pausing regulates mouse embryonic stem cell differentiation.

Melodi Tastemel1, Aishwarya A Gogate2, Venkat S Malladi2

  • 1Cecil H. and Ida Green Center for Reproductive Biology Sciences and Division of Basic Reproductive Biology Research, Department of Obstetrics and Gynecology, University of Texas Southwestern Medical Center, Dallas, TX, USA; Genetics, Development and Diseases Graduate Program, University of Texas Southwestern Medical Center, Dallas, TX, USA.

Stem Cell Research
|November 28, 2017
PubMed
Summary

Embryonic stem cell (ESC) pluripotency requires proper gene regulation. This study shows that disrupting SPT5 impairs RNA polymerase II (Pol II) pausing, hindering ESC differentiation and highlighting Pol II pausing

Keywords:
Global run-on sequencing (GRO-seq)Mouse embryonic stem cellTranscription pausing

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Area of Science:

  • Molecular Biology
  • Developmental Biology
  • Stem Cell Biology

Background:

  • Embryonic stem cell (ESC) pluripotency depends on precise gene expression control.
  • Promoter-proximal RNA polymerase II (Pol II) pausing is implicated in maintaining gene expression readiness.
  • The role of Pol II pausing in ESC pluripotency and differentiation remains to be fully elucidated.

Purpose of the Study:

  • To investigate the function of Pol II pausing in regulating mouse ESC (mESC) pluripotency.
  • To determine the impact of SPT5, a pause-inducing factor, on Pol II pausing and ESC differentiation.

Main Methods:

  • Generation of mESCs with a mutation in the SPT5 gene.
  • Genome-wide analysis of Pol II pausing using genomic techniques.
  • Assessment of ESC differentiation capacity upon disruption of SPT5 function.

Main Results:

  • Mutant SPT5 led to a genome-wide reduction in paused Pol II.
  • A strong correlation was observed between Pol II pausing effects and the local chromatin environment.
  • SPT5-deficient ESCs exhibited impaired differentiation upon withdrawal of self-renewal signals.

Conclusions:

  • Pol II pausing plays a critical role in regulating ESC differentiation.
  • Pol II pausing likely collaborates with epigenetic modifications to control transcription during mESC differentiation.
  • SPT5 is a key regulator of Pol II pausing and subsequent ESC fate decisions.