CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming.
CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming.
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DOI：
10.1016/j.stem.2018.09.003
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发表时间：
2018-11-01
期刊：
影响因子：
23.9
通讯作者：
Qi LS
中科院分区：
文献类型：
--
作者：
Liu Y;Yu C;Daley TP;Wang F;Cao WS;Bhate S;Lin X;Still C 2nd;Liu H;Zhao D;Wang H;Xie XS;Ding S;Wong WH;Wernig M;Qi LS
全面鉴定可以指定神经元命运的因素可以为谱系规范和重编程提供有价值的见解，但系统地询问转录因子及其彼此之间的相互作用已被证明在技术上具有挑战性。我们开发了一种 CRISPR 激活 (CRISPRa) 方法来系统地识别神经元命运规范的调节因子。我们通过胚胎干细胞中的混合 CRISPRa 筛选激活了所有内源转录因子和其他调节因子的表达，揭示了包括表观遗传调节因子（例如 Ezh2）在内的可以诱导神经元命运的基因。基于 CRISPR 的系统性因子对激活使我们能够生成神经元分化的遗传相互作用图谱，并确认顶级个体和组合命中是神经元命运的真正诱导物。几个因子对可以直接将成纤维细胞重编程为神经元，这些神经元与内源性神经元共享相似的转录程序。这项研究为系统鉴定驱动细胞命运获得的基因提供了一种公正的发现方法。 Liu 及其同事开发了一种高通量 CRISPR 激活筛选方法，以系统地识别可有效促进 ESC 神经元命运的转录因子。其中一些单个和成对因子可以进一步将成纤维细胞重新编程为神经元，这表明这种方法可能对工程细胞谱系具有广泛的用途。
Comprehensive identification of factors that can specify neuronal fate could provide valuable insights into lineage specification and reprogramming, but systematic interrogation of transcription factors, and their interactions with each other, has proven technically challenging. We developed a CRISPR activation (CRISPRa) approach to systematically identify regulators of neuronal fate specification. We activated expression of all endogenous transcription factors and other regulators via a pooled CRISPRa screen in embryonic stem cells, revealing genes including epigenetic regulators such as Ezh2 that can induce neuronal fate. Systematic CRISPR-based activation of factor pairs allowed us to generate a genetic interaction map for neuronal differentiation, with confirmation of top individual and combinatorial hits as bona fide inducers of neuronal fate. Several factor pairs could directly reprogram fibroblasts into neurons, which shared similar transcriptional programs with endogenous neurons. This study provides an unbiased discovery approach for systematic identification of genes that drive cell fate acquisition. Liu and colleagues developed a high-throughput CRISPR activation screening approach to systematically identify transcription factors that efficiently promote neuronal fate from ESCs. Some of these single and pairwise factors can further reprogram fibroblasts into neurons, showing that this approach may have broad utility for engineering cell lineages.
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影响因子：
64.8
作者：
Konermann S;Brigham MD;Trevino AE;Joung J;Abudayyeh OO;Barcena C;Hsu PD;Habib N;Gootenberg JS;Nishimasu H;Nureki O;Zhang F
通讯作者：
Zhang F
DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
Nanog在表皮干细胞增殖和分化过程中通过β-catenin磷酸化下调Wnt信号通路。
影响因子：
7.5
作者：
Cheng P;Sun X;Yin D;Xu F;Yang K;Qin L;Dong Y;Guo F;Chen A;Zhang W;Huang H
通讯作者：
Huang H
影响因子：
64.5
作者：
Dixit, Atray;Pamas, Oren;Li, Biyu;Chen, Jenny;Fulco, Charles P.;Jerby-Amon, Livnat;Marjanovic, Nemanja D.;Dionne, Danielle;Burks, Tyler;Raychowdhury, Raktima;Adamson, Britt;Norman, Thomas M.;Lander, Eric S.;Weissman, Jonathan S.;Friedman, Nir;Regev, Aviv
通讯作者：
Regev, Aviv
影响因子：
23.9
作者：
Black, Joshua B.;Adler, Andrew F.;Wang, Hong-Gang;D'Ippolito, Anthony M.;Hutchinson, Hunter A.;Reddy, Timothy E.;Pitt, Geoffrey S.;Leong, Kam W.;Gersbach, Charles A.
通讯作者：
Gersbach, Charles A.