Table1_A One-step strategy to target essential factors with auxin-inducible degron system in mouse embryonic stem cells.DOCX

The self-renewal and pluripotency of embryonic stem cells (ESCs) are conferred by networks including transcription factors and histone modifiers. The Auxin-inducible degron (AID) system can rapidly and reversibly degrade its target proteins and is becoming a powerful tool to explore novel function of key pluripotent and histone modifier genes in ESCs. However, the low biallelic tagging efficiency and a basal degradation level of the current AID systems deem it unsuitable to target key pluripotent genes with tightly controlled expression levels. Here, we develop a one-step strategy to successfully target and repress the endogenous pluripotent genes in mouse ESCs and replace their expression with AID fused transgenes. Therefore, this work provides an efficient way for employing the AID system to uncover novel function of essential pluripotent and chromatin modifier genes in ESCs.

胚胎干细胞（ESCs）的自我更新和多能性由包括转录因子和组蛋白修饰酶在内的网络所赋予。由生长素诱导的降解子（AID）系统能够快速且可逆地降解其目标蛋白，并正逐渐成为探索ESCs中关键多能性和组蛋白修饰基因新功能的强大工具。然而，当前AID系统在双等位基因标记效率低和基础降解水平下，被认为不适用于靶向具有严格控制表达水平的关键多能性基因。在此，我们开发了一种一步法策略，成功靶向并抑制小鼠ESCs中的内源性多能性基因，并用AID融合的转基因替换其表达。因此，本研究为高效利用AID系统揭示ESCs中关键多能性和染色质修饰基因的新功能提供了一种有效途径。