全基因组筛选揭示了 2 细胞样细胞状态的新调控因子。
A genome-wide screen reveals new regulators of the 2-cell-like cell state.
机构信息
Epigenetics and Cell Fate, Université Paris Cité, CNRS, Paris, France.
Joint AZ CRUK Functional Genomics Centre, The Milner Therapeutics Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, UK.
出版信息
Nat Struct Mol Biol. 2023 Aug;30(8):1105-1118. doi: 10.1038/s41594-023-01038-z. Epub 2023 Jul 24.
In mammals, only the zygote and blastomeres of the early embryo are totipotent. This totipotency is mirrored in vitro by mouse '2-cell-like cells' (2CLCs), which appear at low frequency in cultures of embryonic stem cells (ESCs). Because totipotency is not completely understood, we carried out a genome-wide CRISPR knockout screen in mouse ESCs, searching for mutants that reactivate the expression of Dazl, a gene expressed in 2CLCs. Here we report the identification of four mutants that reactivate Dazl and a broader 2-cell-like signature: the E3 ubiquitin ligase adaptor SPOP, the Zinc-Finger transcription factor ZBTB14, MCM3AP, a component of the RNA processing complex TREX-2, and the lysine demethylase KDM5C. All four factors function upstream of DPPA2 and DUX, but not via p53. In addition, SPOP binds DPPA2, and KDM5C interacts with ncPRC1.6 and inhibits 2CLC gene expression in a catalytic-independent manner. These results extend our knowledge of totipotency, a key phase of organismal life.
在哺乳动物中，只有受精卵和早期胚胎的卵裂球具有全能性。这种全能性在体外由小鼠“2 细胞样细胞”（2CLCs）反映出来，这些细胞在胚胎干细胞（ESCs）的培养中以低频率出现。由于全能性尚未完全理解，我们在小鼠 ESCs 中进行了全基因组 CRISPR 敲除筛选，寻找能重新激活 Dazl 表达的突变体，Dazl 是一种在 2CLCs 中表达的基因。在这里，我们报告了四个重新激活 Dazl 和更广泛的 2 细胞样特征的突变体的鉴定：E3 泛素连接酶接头 SPOP、锌指转录因子 ZBTB14、MCM3AP，是 RNA 加工复合物 TREX-2 的一个组成部分，以及赖氨酸去甲基酶 KDM5C。这四个因素都在 DPPA2 和 DUX 的上游起作用，但不通过 p53。此外，SPOP 结合 DPPA2，而 KDM5C 与 ncPRC1.6 相互作用，并以非催化依赖性方式抑制 2CLC 基因表达。这些结果扩展了我们对全能性的认识，全能性是生物体生命的关键阶段。
Zotero 插件