CRISPR-Cas9 screen in human embryonic stem cells to identify genes required for neural differentiation.
CRISPR-Cas9 screen in human embryonic stem cells to identify genes required for neural differentiation.
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DOI：
10.1016/j.xpro.2022.101682
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发表时间：
2022-12-16
期刊：
影响因子：
--
通讯作者：
Yu, Hongtao
中科院分区：
文献类型：
--
作者：
Qi, Shutao;Sivakumar, Sushama;Yu, Hongtao
人类胚胎干细胞(HESCs)在培养过程中不断自我更新，并可被诱导分化为多种细胞类型，包括神经前体细胞(NPC)。在这里，我们提出了一种在hESCs中进行CRISPR-Cas9筛选的方案，以确定在鼻咽癌形成过程中促进SOX1表达的调控因子。该筛选方法可适用于其他终点报告者，用于鉴定与hESCs向其他细胞谱系承诺有关的基因。有关本议定书的使用和执行的完整细节，请参阅。神经分化期间在人ESCs中进行全基因组CRISPR-Cas9筛查流式细胞术为高通量遗传筛查分离感兴趣的细胞内源性蛋白的抗体标记绕过了对报告细胞系的需要该协议通常适用于基于流式细胞术的CRISPR筛查出版者注：进行任何实验方案都需要遵守当地机构的实验室安全和伦理指南。人类胚胎干细胞(HESCs)在培养过程中不断自我更新，并可被诱导分化为多种细胞类型，包括神经前体细胞(NPC)。在这里，我们提出了一种在hESCs中进行CRISPR-Cas9筛选的方案，以确定在鼻咽癌形成过程中促进SOX1表达的调控因子。该筛选方法可适用于其他终点报告者，用于鉴定与hESCs向其他细胞谱系承诺有关的基因。
Human embryonic stem cells (hESCs) continuously self-renew in culture and can be induced to differentiate into multiple cell types, including neural progenitor cells (NPCs). Here, we present a protocol to perform a CRISPR-Cas9 screen in hESCs to identify regulators that promote SOX1 expression during NPC formation. This screening protocol can be adapted with other endpoint reporters for the identification of genes involved in the commitment of hESCs to other cell lineages. For complete details on the use and execution of this protocol, please refer to. Genome-wide CRISPR-Cas9 screen in human ESCs during neural differentiation Flow cytometry to isolate cells of interest for high-throughput genetic screens Antibody labeling of endogenous proteins bypasses the need for reporter cell lines The protocol is generally applicable to flow-cytometry-based CRISPR screens Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Human embryonic stem cells (hESCs) continuously self-renew in culture and can be induced to differentiate into multiple cell types, including neural progenitor cells (NPCs). Here, we present a protocol to perform a CRISPR-Cas9 screen in hESCs to identify regulators that promote SOX1 expression during NPC formation. This screening protocol can be adapted with other endpoint reporters for the identification of genes involved in the commitment of hESCs to other cell lineages.
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影响因子：
8.8
作者：
Sivakumar S;Qi S;Cheng N;Sathe AA;Kanchwala M;Kumar A;Evers BM;Xing C;Yu H
通讯作者：
Yu H
基因组尺度CRISPR-CAS9敲除和转录激活筛选。
影响因子：
14.8
作者：
Joung J;Konermann S;Gootenberg JS;Abudayyeh OO;Platt RJ;Brigham MD;Sanjana NE;Zhang F
通讯作者：
Zhang F
DOI：
10.1126/science.1247005
发表时间：
2014-01-03
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Shalem O;Sanjana NE;Hartenian E;Shi X;Scott DA;Mikkelson T;Heckl D;Ebert BL;Root DE;Doench JG;Zhang F
通讯作者：
Zhang F
影响因子：
12.3
作者：
Li W;Xu H;Xiao T;Cong L;Love MI;Zhang F;Irizarry RA;Liu JS;Brown M;Liu XS
通讯作者：
Liu XS