干细胞多能性获得及丧失与全基因组转录水平及染色质结构密切相关，受表观遗传修饰及识别蛋白调控。甲基结合蛋白（MBD）是重要的识别蛋白，在染色质重构和基因表达调控中发挥重要作用。本组前期发现甲基结合蛋白1（Mbd1）对小鼠胚胎干细胞多能性维持具有重要作用，但其对人胚胎干细胞的作用及影响多能性的机制尚不清楚。本项目将使用Crispr/Cas9方法获得MBD1敲除的人胚胎干细胞，并检测其对人胚胎干细胞多能性的影响；使用体外诱导多能干细胞技术研究MBD1在小鼠和人体细胞重编程过程中的功能；利用基因亚型荧光标记技术探讨不同亚型MBD1对胚胎干细胞多能性的影响及机制；利用干细胞体外分化技术研究胚胎/诱导多能干细胞定向分化为神经前体细胞中MBD1的作用及机制。本研究预期阐明胚胎干细胞分化（神经前体细胞）和诱导多能干细胞获得过程中MBD1的功能及调控机制，补充干细胞多能性调控中表观遗传学的理论。

During the gain and loss of stem cell pluripotency, the whole genome transcription and chromatin remolding occur. The epigenetic information of 5-methylcytosine (5mC) in mammalian cells can be translated by cytosine modification readers, such as the methyl-CpG binding domain (MBD) proteins. The interplay between 5mC and MBDs plays a major role in chromatin remodeling and gene expression regulation, but the role of the interplay is still unclear in the differentiation of embryonic stem cells and induction of pluripotent cell. In our previous study, we showed that the Mbd1 is very crucial for the pluripotency maintenance of mouse embryonic stem cells. In this study, we will establish a human MBD1 knockout cell line using Crispr/Cas9 and study the role of MBD1 on the pluripotency maintenance of human embryonic stem cells. Then, we will investigate the role of Mbd1 during the reprogramming of human and mouse somatic cells using induced pluripotent stem cell technology. Moreover, we will focus on the role of Mbd1 isoform in the gain and loss of pluripotency in vitro and in vivo using gene isoform fluorescently labeling technology. Finally, we will investigate the role of Mbd1 in the differentiation of mouse and human pluripotent cells to neural stem/progenitor cells. The outcome of this study will not only elucidate the function and mechanism of Mbd1 in embryonic stem cell differentiation (to neural stem/progenitor cells) and induced pluripotent stem cell reprogramming, but also provide more evidence for the epigenetic regulation of stem cell pluripotency.