诱导降解系统揭示 PRDM14 在人原始生殖细胞命运决定中的关键作用。
A critical role of PRDM14 in human primordial germ cell fate revealed by inducible degrons.
机构信息
Wellcome Trust/Cancer Research UK Gurdon Institute, Henry Wellcome Building of Cancer and Developmental Biology, Cambridge, CB2 1QN, UK.
Physiology, Development and Neuroscience Department, University of Cambridge, Cambridge, CB2 3EL, UK.
出版信息
Nat Commun. 2020 Mar 9;11(1):1282. doi: 10.1038/s41467-020-15042-0.
PRDM14 is a crucial regulator of mouse primordial germ cells (mPGCs), epigenetic reprogramming and pluripotency, but its role in the evolutionarily divergent regulatory network of human PGCs (hPGCs) remains unclear. Besides, a previous knockdown study indicated that PRDM14 might be dispensable for human germ cell fate. Here, we decided to use inducible degrons for a more rapid and comprehensive PRDM14 depletion. We show that PRDM14 loss results in significantly reduced specification efficiency and an aberrant transcriptome of hPGC-like cells (hPGCLCs) obtained in vitro from human embryonic stem cells (hESCs). Chromatin immunoprecipitation and transcriptomic analyses suggest that PRDM14 cooperates with TFAP2C and BLIMP1 to upregulate germ cell and pluripotency genes, while repressing WNT signalling and somatic markers. Notably, PRDM14 targets are not conserved between mouse and human, emphasising the divergent molecular mechanisms of PGC specification. The effectiveness of degrons for acute protein depletion is widely applicable in various developmental contexts.
PRDM14 是小鼠原始生殖细胞（mPGCs）、表观遗传重编程和多能性的关键调节因子，但它在人类原始生殖细胞（hPGCs）进化上不同的调节网络中的作用尚不清楚。此外，之前的敲低研究表明，PRDM14 对于人类生殖细胞命运可能不是必需的。在这里，我们决定使用诱导降解结构域进行更快速和全面的 PRDM14 耗竭。我们发现 PRDM14 的缺失导致体外从人类胚胎干细胞（hESCs）获得的 hPGC 样细胞（hPGCLCs）的特异性效率显著降低，且转录组异常。染色质免疫沉淀和转录组分析表明，PRDM14 与 TFAP2C 和 BLIMP1 合作上调生殖细胞和多能性基因，同时抑制 WNT 信号和体细胞标记。值得注意的是，PRDM14 的靶标在小鼠和人类之间没有保守，强调了 PGC 特化的分子机制的分歧。降解结构域用于急性蛋白质耗竭的有效性在各种发育背景中广泛适用。
Zotero 插件