染色质的三维空间结构中增强子-启动子间的远距离互作对基因的转录过程具有重要的调控作用。目前，不同的互作类型对Sox2基因转录调控的分子机制及其对胚胎干细胞多能性维持的生物学效应还不清楚。本项目在前期研究中鉴定到Sox2基因相关的三个不同类型增强子-启动子的远距离互作，利用CRISPR/Cas9在胚胎干细胞中构建了增强子敲除细胞株；利用RNA-Seq鉴定了不同互作可能影响到的差异表达基因，并分别对干细胞多能性、分化潜能及相关表型进行了研究。未来拟利用高效Hi-C技术，构建不同细胞的染色质三维互作图谱，以Sox2基因为核心构建三维调控网络，揭示不同互作类型对其转录调控及三维空间构象的影响；与此同时，本项目将联合分析三维基因组和其他多组学数据，探究其与胚胎干细胞多能性之间的关系。这将为基因的时空特异性转录调控研究提供新的思路，有望推进三维基因组学在关键基因或重要转录因子功能研究中的应用。

In embryonic stem cells, three-dimensional (3D) conformation of genome and distal enhancer-promoter interactions have critical impact on Sox2 gene regulation. At present, its detailed molecular mechanism and biological effect on pluripotency maintenance of embryonic stem cells (ESCs) are still unclear. With three different kinds of identified distal interactions, we constructed enhancer knockout cell lines in ESCs using CRISPR/Cas9. We get the differentiated expression gene list by using RNA-Seq and further study potential effects on pluripotency and differentiation ability in ESCs. In the future, we plan use efficient Hi-C to reconstruct the 3D interactome in each cells. And will use Sox2 gene as the core factor to rebuild each individual network and do the comparison by using differentiated DNA-Loops. Furthermore, these will be combined with other -omics datasets to analyze the biological effect and molecular mechanism of different interactions. This project provides a new idea on spatiotemporal gene expression researches and will facilitate to apply 3D genome information on key gene/transcription factor function studies.