Nuclear architecture underlies the transcriptional programs within the cell to establish cell identity. As previously demonstrated, long-range chromatin interactions of the Oct4 distal enhancer (DE) are correlated with active transcription in naïve state embryonic stem cells. Here, we identify and characterize extreme long-range interactions of the Oct4 DE through a novel CRISPR labeling technique we developed and chromosome conformation capture to identify lethal giant larvae 2 (Llgl2) and growth factor receptor-bound protein 7 (Grb7) as putative functional interacting target genes in different chromosomes. We show that the Oct4 DE directly regulates expression of Llgl2 and Grb7 in addition to Oct4. Expression of Llgl2 and Grb7 closely correlates with the pluripotent state, where knock down of either result in loss of pluripotency, and overexpression enhances somatic cell reprogramming. We demonstrated that biologically important interactions of the Oct4 DE can occur at extreme distances that are necessary for the maintenance of the pluripotent state.

核结构是细胞内转录程序建立细胞身份的基础。如前所述， Oct4远端增强子 (DE) 的长程染色质相互作用与幼稚状态胚胎干细胞中的活性转录相关。在这里，我们通过我们开发的新型 CRISPR 标记技术和染色体构象捕获来识别和表征Oct4 DE 的极端长程相互作用，以识别致命巨型幼虫 2 ( Llgl2 ) 和生长因子受体结合蛋白 7 ( Grb7 ) 作为假定的功能不同染色体上相互作用的靶基因。我们发现除了Oct4之外， Oct4 DE 还直接调节Llgl2和Grb7的表达。 Llgl2和Grb7的表达与多能状态密切相关，其中任一个的敲低都会导致多能性的丧失，而过度表达会增强体细胞重编程。我们证明了Oct4 DE 的生物学上重要的相互作用可以在维持多能状态所必需的极端距离处发生。