The progression from naive through formative to primed in vitro pluripotent stem cell states recapitulates epiblast development in vivo during the peri-implantation period of mouse embryo development. Activation of the de novo DNA methyltransferases and reorganization of transcriptional and epigenetic landscapes are key events that occur during these pluripotent state transitions. However, the upstream regulators that coordinate these events are relatively underexplored. Here, using Zfp281 knockout mouse and degron knockin cell models, we identify the direct transcriptional activation of Dnmt3a/3b by ZFP281 in pluripotent stem cells. Chromatin co-occupancy of ZFP281 and DNA hydroxylase TET1, which is dependent on the formation of R-loops in ZFP281-targeted gene promoters, undergoes a “high-low-high” bimodal pattern regulating dynamic DNA methylation and gene expression during the naive-formative-primed transitions. ZFP281 also safeguards DNA methylation in maintaining primed pluripotency. Our study demonstrates a previously unappreciated role for ZFP281 in coordinating DNMT3A/3B and TET1 functions to promote pluripotent state transitions.

体外多能干细胞状态从幼稚到形成再到启动的过程概括了小鼠胚胎发育的围植入期体内外胚层的发育。从头DNA 甲基转移酶的激活以及转录和表观遗传景观的重组是这些多能状态转换期间发生的关键事件。然而，协调这些事件的上游监管机构的探索相对不足。在这里，使用Zfp281敲除小鼠和 degron 敲入细胞模型，我们鉴定了ZFP281 在多能干细胞中对Dnmt3a/3b的直接转录激活。ZFP281 和 DNA 羟化酶 TET1 的染色质共占用依赖于 ZFP281 靶向基因启动子中 R 环的形成，在初始阶段经历“高-低-高”双峰模式调节动态 DNA 甲基化和基因表达。形成引发的转变。ZFP281 还可以保护 DNA 甲基化以维持引发的多能性。我们的研究表明，ZFP281 在协调 DNMT3A/3B 和 TET1 功能以促进多能状态转换方面发挥着先前未被认识到的作用。