Pluripotency-State-Dependent Role of Dax1 in Embryonic Stem Cells Self-Renewal

Jianrong He; Yuda Cheng; Yan Ruan; Jiali Wang; Yanping Tian; Jiaqi Wang; Fengsheng Wang; Chen Zhang; Yixiao Xu; Lianlian Liu; Meng Yu; Jiangjun Wang; Binyu Zhao; Yue Zhang; Yi Yang; Gaoke Liu; Wei Wu; Ping He; Jiaxiang Xiong; He Huang; Junlei Zhang; Rui Jian

doi:10.1155/2021/5522723

Pluripotency-State-Dependent Role of Dax1 in Embryonic Stem Cells Self-Renewal

Stem Cells Int. 2021 Jul 10:2021:5522723. doi: 10.1155/2021/5522723. eCollection 2021.

Authors

Jianrong He^{1

2}, Yuda Cheng², Yan Ruan², Jiali Wang², Yanping Tian², Jiaqi Wang², Fengsheng Wang², Chen Zhang², Yixiao Xu^{2

3}, Lianlian Liu², Meng Yu², Jiangjun Wang², Binyu Zhao², Yue Zhang^{2

3}, Yi Yang⁴, Gaoke Liu², Wei Wu⁵, Ping He⁶, Jiaxiang Xiong⁴, He Huang¹, Junlei Zhang², Rui Jian²

Affiliations

¹ Department of Anesthesiology, The Second Affiliated Hospital, Chongqing Medical University, Chongqing 400010, China.
² Laboratory of Stem Cell & Developmental Biology, Department of Histology and Embryology, Army Medical University, Chongqing 400038, China.
³ Southwest Hospital/Southwest Eye Hospital, The First Hospital Affiliated to Army Medical University, Chongqing 400038, China.
⁴ Experimental Center of Basic Medicine, College of Basic Medical Sciences, Army Medical University, Chongqing 400038, China.
⁵ Thoracic Surgery Department, Southwest Hospital, The First Hospital Affiliated to Army Medical University, Chongqing 400038, China.
⁶ Cardiac Surgery Department, Southwest Hospital, The First Hospital Affiliated to Army Medical University, Chongqing 400038, China.

Abstract

Dax1(also known as Nr0b1) is regarded as an important component of the transcription factor network in mouse embryonic stem cells (ESCs). However, the role and the molecular mechanism of Dax1 in the maintenance of different pluripotency states are poorly understood. Here, we constructed a stable Dax1 knockout (KO) cell line using the CRISPR/Cas9 system to analyze the precise function of Dax1. We reported that 2i/LIF-ESCs had significantly lower Dax1 expression than LIF/serum-ESCs. Dax1KO ES cell lines could be established in 2i/LIF and their pluripotency was confirmed. In contrast, Dax1-null ESCs could not be continuously passaged in LIF/serum due to severe differentiation and apoptosis. In LIF/serum, the activities of the Core module and Myc module were significantly reduced, while the PRC2 module was activated after Dax1KO. The expression of most proapoptotic genes and lineage-commitment genes were drastically increased, while the downregulated expression of antiapoptotic genes and many pluripotency genes was observed. Our research on the pluripotent state-dependent role of Dax1 provides clues to understand the molecular regulation mechanism at different stages of early embryonic development.