PRC2 Facilitates the Regulatory Topology Required for Poised Enhancer Function during Pluripotent Stem Cell Differentiation
PRC2 Facilitates the Regulatory Topology Required for Poised Enhancer Function during Pluripotent Stem Cell Differentiation
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DOI：
10.1016/j.stem.2017.02.004
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发表时间：
2017-05-04
期刊：
影响因子：
23.9
通讯作者：
Rada-Iglesias, Alvaro
中科院分区：
文献类型：
--
作者：
Cruz-Molina, Sara;Respuela, Patricia;Rada-Iglesias, Alvaro
在多能干细胞中由H3 K27 me 3标记的定位增强子已经涉及在胚胎干细胞（ESC）分化期间建立体细胞表达程序。然而，潜在增强子的功能相关性和作用机制仍然未知。使用CRISPR/Cas9技术来设计精确的遗传缺失，我们证明了平衡的增强子对于诱导主要的前神经调节因子是必要的。有趣的是，环状染色体构象捕获测序（4C-seq）显示，平衡增强子已经以多梳抑制复合物2（PRC 2）依赖性方式与ESC中的靶基因建立了物理相互作用。PRC 2的丢失不会激活未分化ESC中的平衡增强子或诱导其推定的靶基因;然而，在分化ESC中PRC 2的丢失严重且特异性地损害代表平衡增强子靶的主要前神经基因的诱导。总的来说，我们的工作阐明了一个意想不到的功能polycomb蛋白在促进神经诱导赋予主要的前神经基因座与一个许可的监管拓扑结构。
Poised enhancers marked by H3K27me3 in pluripotent stem cells have been implicated in the establishment of somatic expression programs during embryonic stem cell (ESC) differentiation. However, the functional relevance and mechanism of action of poised enhancers remain unknown. Using CRISPR/Cas9 technology to engineer precise genetic deletions, we demonstrate that poised enhancers are necessary for the induction of major anterior neural regulators. Interestingly, circularized chromosome conformation capture sequencing (4C-seq) shows that poised enhancers already establish physical interactions with their target genes in ESCs in a polycomb repressive complex 2 (PRC2)-dependent manner. Loss of PRC2 does not activate poised enhancers or induce their putative target genes in undifferentiated ESCs; however, loss of PRC2 in differentiating ESCs severely and specifically compromises the induction of major anterior neural genes representing poised enhancer targets. Overall, our work illuminates an unexpected function for polycomb proteins in facilitating neural induction by endowing major anterior neural loci with a permissive regulatory topology.