Open AccessXist Repression Shows Time‐Dependent Effects on the Reprogramming of Female Somatic Cells to Induced Pluripotent Stem Cells
Author(s) -
Chen Qi,
Gao Shuai,
He Wenteng,
Kou Xiaochen,
Zhao Yanhong,
Wang Hong,
Gao Shaorong
Publication year - 2014
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1002/stem.1775
Subject(s) - xist , reprogramming , biology , induced pluripotent stem cell , somatic cell , x inactivation , psychological repression , microbiology and biotechnology , gene knockdown , x chromosome , stem cell , genetics , embryonic stem cell , cell , gene expression , cell culture , gene
Abstract Although the reactivation of silenced X chromosomes has been observed as part of the process of reprogramming female somatic cells into induced pluripotent stem cells (iPSCs), it remains unknown whether repression of the X‐inactive specific transcript ( Xist ) can greatly enhance female iPSC induction similar to that observed in somatic cell nuclear transfer studies. In this study, we discovered that the repression of Xist plays opposite roles in the early and late phases of female iPSCs induction. Our results demonstrate that the downregulation of Xist by an isopropyl β‐ d ‐1‐thiogalactopyranoside (IPTG)‐inducible short hairpin RNA (shRNA) system can greatly impair the mesenchymal‐to‐epithelial transition (MET) in the early phase of iPSC induction but can significantly promote the transition of pre‐iPSCs to iPSCs in the late phase. Furthermore, we demonstrate that although the knockdown of Xist did not affect the H3K27me3 modification on the X chromosome, macroH2A was released from the inactivated X chromosome (Xi). This enables the X chromosome silencing to be a reversible event. Moreover, we demonstrate that the supplementation of vitamin C (Vc) can augment and stabilize the reversible X chromosome by preventing the relocalization of macroH2A to the Xi. Therefore, our study reveals an opposite role of Xist repression in the early and late stages of reprogramming female somatic cells to pluripotency and demonstrates that the release of macroH2A by Xist repression enables the transition from pre‐iPSCs to iPSCs. S tem C ells 2014;32:2642–2656