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A novel approach to differentiate rat embryonic stem cells in vitro reveals a role for RNF12 in activation of X chromosome inactivation

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  • Aristea Magaraki
  • Agnese Loda
  • Cristina Gontan
  • Sarra Merzouk
  • Esther Sleddens-Linkels
  • Stephen Meek
  • Willy M. Baarends
  • Thomas Burdon
  • Joost Gribnau

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Original languageEnglish
JournalScientific Reports
Early online date15 Apr 2019
Publication statusE-pub ahead of print - 15 Apr 2019


X chromosome inactivation (XCI) is a eutherian specific, developmentally regulated process relying on several mechanisms including antisense transcription, non-coding RNA-mediated silencing, and recruitment of chromatin remodeling complexes. In vitro modeling of XCI, through differentiation of embryonic stem cells (ESCs), provides a powerful tool to study the dynamics of XCI, overcoming the need for embryos, and facilitating genetic modification of key regulatory players. However, to date, robust initiation of XCI in vitro has been mostly limited to mouse pluripotent stem cells. Here, we adapted eXisting protocols to establish a novel monolayer differentiation protocol for rat ESCs to study XCI. We show that differentiating rat ESCs properly downregulate pluripotency factor genes, and present female specific Xist RNA accumulation and silencing of X-linked genes. We also demonstrate that
RNF12 seems to be an important player in regulation of initiation of XCI in rat, acting as an Xist activator. Our work provides the basis to investigate the mechanisms directing the XCI process in a model organism different from the mouse.

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