A genome-wide CRISPR-Cas9 knockout screen identifies essential and growth-restricting genes in human trophoblast stem cells.
A genome-wide CRISPR-Cas9 knockout screen identifies essential and growth-restricting genes in human trophoblast stem cells.
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DOI：
10.1038/s41467-022-30207-9
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发表时间：
2022-05-10
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
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作者：
最近人类滋养细胞干细胞（hTSCs）的衍生提供了一个可扩展的人类胎盘发育的体外模型系统，但hTSC身份的分子调节因子迄今尚未系统地探索。在这里，我们利用全基因组CRISPR-Cas9敲除筛选来全面鉴定hTSCs中的必需和生长限制基因。通过将我们的数据与在其他细胞类型中进行的类似基因筛选的数据以及早期人类胚胎的基因表达数据交叉对照，我们定义了htsc特异性和富集的调节因子。这些包括已建立的和以前未表征的滋养细胞调节因子，如ARID3A， GATA2和TEAD1（必需），以及GCM1， PTPN14和TET2（生长限制）。染色质可及性、基因表达和全基因组定位数据的综合分析显示，转录因子TEAD1调节hTSCs中许多滋养细胞调节因子的表达。在缺乏TEAD1的情况下，hTSCs不能完全忠实地分化为上皮外滋养细胞（EVT）细胞，而是倾向于向合胞滋养细胞（STB）分化，从而表明该转录因子保护了hTSCs的双潜能谱系。总之，我们的研究为剖析人类胎盘发育和疾病的分子调控提供了宝贵的资源。在这里，作者进行了全基因组CRISPR-Cas9敲除筛选，以系统地鉴定和表征人类滋养细胞中的必需基因和生长限制基因。他们发现TEAD1是一个关键的调节因子，通过调节染色质结构和基因表达，在人类滋养细胞谱系的规范、维持和分化中发挥重要作用。
The recent derivation of human trophoblast stem cells (hTSCs) provides a scalable in vitro model system of human placental development, but the molecular regulators of hTSC identity have not been systematically explored thus far. Here, we utilize a genome-wide CRISPR-Cas9 knockout screen to comprehensively identify essential and growth-restricting genes in hTSCs. By cross-referencing our data to those from similar genetic screens performed in other cell types, as well as gene expression data from early human embryos, we define hTSC-specific and -enriched regulators. These include both well-established and previously uncharacterized trophoblast regulators, such as ARID3A, GATA2, and TEAD1 (essential), and GCM1, PTPN14, and TET2 (growth-restricting). Integrated analysis of chromatin accessibility, gene expression, and genome-wide location data reveals that the transcription factor TEAD1 regulates the expression of many trophoblast regulators in hTSCs. In the absence of TEAD1, hTSCs fail to complete faithful differentiation into extravillous trophoblast (EVT) cells and instead show a bias towards syncytiotrophoblast (STB) differentiation, thus indicating that this transcription factor safeguards the bipotent lineage potential of hTSCs. Overall, our study provides a valuable resource for dissecting the molecular regulation of human placental development and diseases. Here the authors perform a genome-wide CRISPR-Cas9 knockout screen to systematically identify and characterize essential and growth-restricting genes in human trophoblast cells. They identify TEAD1 as a key regulator that plays an important role in the specification, maintenance, and differentiation of the human trophoblast lineage by modulating chromatin architecture and gene expression.
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影响因子：
46.9
作者：
Doench JG;Fusi N;Sullender M;Hegde M;Vaimberg EW;Donovan KF;Smith I;Tothova Z;Wilen C;Orchard R;Virgin HW;Listgarten J;Root DE
通讯作者：
Root DE
DOI：
10.1007/978-1-0716-1908-7_2
发表时间：
2022
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
通讯作者：
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影响因子：
7.7
作者：
Dong, Chen;Beltcheva, Mariana;Theunissen, Thorold W.
通讯作者：
Theunissen, Thorold W.
影响因子：
5.9
作者：
Cinkornpumin, Jessica K.;Kwon, Sin Young;Pastor, William A.
通讯作者：
Pastor, William A.
影响因子：
9.2
作者：
Cockburn, Katie;Biechele, Steffen;Rossant, Janet
通讯作者：
Rossant, Janet