Pooled CRISPR-activation screening coupled with single-cell RNA-seq in mouse embryonic stem cells.
Pooled CRISPR-activation screening coupled with single-cell RNA-seq in mouse embryonic stem cells.
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DOI：
10.1016/j.xpro.2021.100426
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发表时间：
2021-06-18
期刊：
影响因子：
--
通讯作者：
Reik W
中科院分区：
文献类型：
--
作者：
Alda-Catalinas C;Eckersley-Maslin MA;Reik W
CRISPR/Cas9筛选是识别生物过程关键调控因子的有效方法。通过将合并的CRISPR/Cas9筛选与单细胞RNA测序读数相结合，可以全面和大规模地平行评估个体扰动。重要的是，这使得基因功能和调控可以在细胞水平上以无偏的方式进行询问。在这里，我们提出了一种使用10× Genomics scRNA-seq作为读数在小鼠胚胎干细胞中进行合并CRISPR激活筛选的方案。有关本协议生成和使用的完整信息，请参阅。CRISPR/Cas9筛选是鉴定生物学过程关键调控因子的有效方法。CRISPR/Cas9筛选是鉴定生物学过程关键调控因子的有效方法。通过将合并的CRISPR/Cas9筛选与单细胞RNA测序读数相结合，可以全面和大规模地平行评估个体扰动。重要的是，这使得基因功能和调控可以在细胞水平上以无偏的方式进行询问。在这里，我们提出了一种使用10× Genomics scRNA-seq作为读数在小鼠胚胎干细胞中进行合并CRISPR激活筛选的方案。
CRISPR/Cas9 screens are a powerful approach to identify key regulators of biological processes. By combining pooled CRISPR/Cas9 screening with single-cell RNA-sequencing readout, individual perturbations can be assessed in parallel both comprehensively and at scale. Importantly, this allows gene function and regulation to be interrogated at a cellular level in an unbiased manner. Here, we present a protocol to perform pooled CRISPR-activation screens in mouse embryonic stem cells using 10× Genomics scRNA-seq as a readout. For complete information on the generation and use of this protocol, please refer to. Protocol for CRISPRa screens with single-cell readout to interrogate gene function Detailed description of CRISPRa screening procedures in mouse embryonic stem cells Detailed steps on how to construct derived single-cell sgRNA amplicon libraries CRISPR/Cas9 screens are a powerful approach to identify key regulators of biological processes. By combining pooled CRISPR/Cas9 screening with a single-cell RNA-sequencing readout, individual perturbations can be assessed in parallel both comprehensively and at scale. Importantly, this allows gene function and regulation to be interrogated at a cellular level in an unbiased manner. Here, we present a protocol to perform pooled CRISPR-activation screens in mouse embryonic stem cells using 10× Genomics scRNA-seq as a readout.
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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
影响因子：
16.6
作者：
Zheng GX;Terry JM;Belgrader P;Ryvkin P;Bent ZW;Wilson R;Ziraldo SB;Wheeler TD;McDermott GP;Zhu J;Gregory MT;Shuga J;Montesclaros L;Underwood JG;Masquelier DA;Nishimura SY;Schnall-Levin M;Wyatt PW;Hindson CM;Bharadwaj R;Wong A;Ness KD;Beppu LW;Deeg HJ;McFarland C;Loeb KR;Valente WJ;Ericson NG;Stevens EA;Radich JP;Mikkelsen TS;Hindson BJ;Bielas JH
通讯作者：
Bielas JH
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
影响因子：
16.6
作者：
Sanson, Kendall R.;Hanna, Ruth E.;Doench, John G.
通讯作者：
Doench, John G.
影响因子：
48
作者：
Hill, Andrew J.;McFaline-Figueroa, Jose L.;Trapnell, Cole
通讯作者：
Trapnell, Cole