Mapping germ-layer specification preventing genes in hPSCs via genome-scale CRISPR screening.
Mapping germ-layer specification preventing genes in hPSCs via genome-scale CRISPR screening.
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通过基因组规模 CRISPR 筛选绘制胚层规范阻止 hPSC 基因的图谱
DOI：
10.1016/j.isci.2020.101926
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发表时间：
2021-01-22
期刊：
影响因子：
5.8
通讯作者：
Zhang X
中科院分区：
文献类型：
--
作者：
Xu X;Du Y;Ma L;Zhang S;Shi L;Chen Z;Zhou Z;Hui Y;Liu Y;Fang Y;Fan B;Liu Z;Li N;Zhou S;Jiang C;Liu L;Zhang X
了解决定人类多能干细胞（hPSC）进入三个胚层的生物学过程是发育和干细胞生物学的中心问题。在这里，我们用胚层报告基因和诱导型CRISPR/Cas9敲除系统对hPSC进行了基因工程改造，并进行了基因组规模的筛选以确定限制胚层特化的途径。聚集在关键生物学过程中的基因，包括胚胎发育、mRNA加工、代谢和表观遗传调节，集中在多能性和谱系发育的管理中。除了典型的多能性转录因子和信号分子，中内胚层特异性因子的功能丧失导致高级神经外胚层分化，因为它们具有胚层间拮抗作用。关于表观遗传超家族，hPSC中富集的microRNA显示出明确的胚层靶向特异性。胆固醇合成途径通过阻滞神经外胚层特化来维持hPSC。因此，在这项研究中，我们确定了控制hPSC自我更新和三系特化的遗传布线和生物过程的完整景观。谱系报告基因和CRISPR筛选是研究细胞命运的有力工具在hPSC中鉴定谱系特异性预防基因（LPGs）LPGs通过靶向一个或多个胚层保持多能性LPGs聚集成不同的功能模块生物科学;细胞生物学;干细胞研究;发育生物学
Understanding the biological processes that determine the entry of three germ layers of human pluripotent stem cells (hPSCs) is a central question in developmental and stem cell biology. Here, we genetically engineered hPSCs with the germ layer reporter and inducible CRISPR/Cas9 knockout system, and a genome-scale screening was performed to define pathways restricting germ layer specification. Genes clustered in the key biological processes, including embryonic development, mRNA processing, metabolism, and epigenetic regulation, were centered in the governance of pluripotency and lineage development. Other than typical pluripotent transcription factors and signaling molecules, loss of function of mesendodermal specifiers resulted in advanced neuroectodermal differentiation, given their inter-germ layer antagonizing effect. Regarding the epigenetic superfamily, microRNAs enriched in hPSCs showed clear germ layer-targeting specificity. The cholesterol synthesis pathway maintained hPSCs via retardation of neuroectoderm specification. Thus, in this study, we identified a full landscape of genetic wiring and biological processes that control hPSC self-renewal and trilineage specification. Lineage reporter and CRISPR screening are powerful tools for studying cell fates Lineage-specification preventing genes (LPGs) are identified in hPSCs LPGs maintain pluripotency via targeting one or multiple germ layers LPGs are clustered into distinct functional modules Biological Sciences; Cell Biology; Stem Cells Research; Developmental Biology
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一个 iCRISPR 平台，用于在人类多能干细胞中进行快速、可多重、可诱导的基因组编辑。
影响因子：
23.9
作者：
Gonzalez, Federico;Zhu, Zengrong;Shi, Zhong-Dong;Lelli, Katherine;Verma, Nipun;Li, Qing V.;Huangfu, Danwei
通讯作者：
Huangfu, Danwei
影响因子：
48
作者：
Langmead, Ben;Salzberg, Steven L.
通讯作者：
Salzberg, Steven L.
影响因子：
64.5
作者：
Gifford CA;Ziller MJ;Gu H;Trapnell C;Donaghey J;Tsankov A;Shalek AK;Kelley DR;Shishkin AA;Issner R;Zhang X;Coyne M;Fostel JL;Holmes L;Meldrim J;Guttman M;Epstein C;Park H;Kohlbacher O;Rinn J;Gnirke A;Lander ES;Bernstein BE;Meissner A
通讯作者：
Meissner A
影响因子：
4.6
作者：
Lees, Jarmon G.;Gardner, David K.;Harvey, Alexandra J.
通讯作者：
Harvey, Alexandra J.
影响因子：
5.9
作者：
Allison TF;Smith AJH;Anastassiadis K;Sloane-Stanley J;Biga V;Stavish D;Hackland J;Sabri S;Langerman J;Jones M;Plath K;Coca D;Barbaric I;Gokhale P;Andrews PW
通讯作者：
Andrews PW