Fu Yuting, Liu Fangyuan, Cao Shuo, Zhang Jia, Wang Huizhi, Wu Baojiang, Song Yongli, Duo Shuguang, Li Xihe, Bao Siqin
State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, Inner Mongolia University, Hohhot, China.
Institute of Animal Genetic Research of Mongolia Plateau, College of Life Sciences, Inner Mongolia University, Hohhot, China.
Front Cell Dev Biol. 2021 Apr 8;9:655145. doi: 10.3389/fcell.2021.655145. eCollection 2021.
3-hydroxybutyrate dehydrogenase-2 (), a short-chain dehydrogenase, catalyzes a rate-limiting step in the biogenesis of the mammalian siderophore, playing a key role in iron homeostasis, energy metabolism and apoptosis. However, the function of in embryonic stem cells (ESCs) remains unknown. To gain insights into the role of on pluripotency and cell fate decisions of mouse ESCs, we generated homozygous knockout lines for both mouse advanced embryonic stem cell (ASC) and ESC using CRISPR/Cas9 genome editing technology. deficiency in both ASCs and ESCs had no effect on expression of core pluripotent transcription factors and alkaline phosphatase activity, suggesting dispensability of for self-renewal and pluripotency of ESCs. Interestingly, cells with deficiency exhibited potency of endoderm differentiation ; with upregulated endoderm associated genes revealed by RNA-seq and RT-qPCR. We further demonstrate that loss inhibited expression of multiple methyltransferases (DNMTs) at both RNA and protein level, suggesting that may be essentially required to maintain DNA methylation in ASCs and ESCs. Overall, this study provides valuable data and resources for understanding how regulate earliest cell fate decision and DNA methylation in ASCs/ESCs.
3-羟基丁酸脱氢酶-2（），一种短链脱氢酶，催化哺乳动物铁载体生物合成中的限速步骤，在铁稳态、能量代谢和细胞凋亡中起关键作用。然而，其在胚胎干细胞（ESC）中的功能仍不清楚。为了深入了解其对小鼠胚胎干细胞多能性和细胞命运决定的作用，我们使用CRISPR/Cas9基因组编辑技术为小鼠高级胚胎干细胞（ASC）和胚胎干细胞生成了纯合敲除系。ASC和ESC中的缺陷对核心多能转录因子的表达和碱性磷酸酶活性没有影响，表明其对胚胎干细胞的自我更新和多能性并非必需。有趣的是，有缺陷的细胞表现出内胚层分化的潜能；RNA测序和逆转录定量聚合酶链反应显示内胚层相关基因上调。我们进一步证明，的缺失在RNA和蛋白质水平上均抑制了多种甲基转移酶（DNMT）的表达，表明可能是维持ASC和ESC中DNA甲基化所必需的。总体而言，本研究为理解如何调节ASC/ESC中最早的细胞命运决定和DNA甲基化提供了有价值的数据和资源。
