Live Cell Imaging of the Nascent Inactive X Chromosome during the Early Differentiation Process of Naive ES Cells towards Epiblast Stem Cells

• DOI: 10.1371/journal.pone.0116109
• Publication Date: 2014-12-29T18:57:49Z
• Authors: Aurélia Guyochin; Sylvain Maenner; Erin Tsi-Jia Chu; Asma Hentati; Mikael Attia; Philip Avner; Philippe Clerc
• Keywords: Epiblast; XIST; PRC2

Random X-chromosome inactivation ensures dosage compensation in mammals through the transcriptional silencing of one two X chromosomes present each female cell. Silencing is initiated differentiating epiblast mouse embryos coating nascent inactive chromosome by non-coding RNA Xist, which subsequently recruits Polycomb Complex PRC2 leading to histone H3-K27 methylation. Here we examined ES cells early steps transition from naive towards stem as a model for inducing vitro. We show that these conditions efficiently induce random XCI. Importantly, transient phase this differentiation pathway, both are coated with Xist up 15% XX cells. In an attempt determine dynamics process, designed strategy aimed at visualizing live generated transgenic expressing component Ezh2 fused fluorescent protein Venus. The fusion was expressed sub-physiological levels and located nuclei Upon cell fate, Venus-fluorescent territories appearing interphase were identified their association RNA. Imaging Ezh2-Venus 24 hours during process showed survival some domains surprising across division course process. Our data reveal suggests possibility large plasticity chromosome.