用于在 HeLa 细胞中进行全基因组 CRISPR 敲除筛选细菌细胞毒素的方案。
Protocol for genome-wide CRISPR knockout screens of bacterial cytotoxins in HeLa cells.
机构信息
College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, China; Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang 310024, China; Center for Infectious Disease Research, Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang 310024, China.
Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang 310024, China; Center for Infectious Disease Research, Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang 310024, China.
出版信息
STAR Protoc. 2022 Jul 31;3(3):101595. doi: 10.1016/j.xpro.2022.101595. eCollection 2022 Sep 16.
CRISPR screening is a powerful tool to identify host factors for pathogenic agents including viruses and bacterial toxins. Here, we present a protocol to conduct a genome-scale CRISPR screen on HeLa cells for host factors involved in the toxin action of TcdB4. We describe in detail how to prepare the library, set up the screen, obtain the gene sequences, and analyze the results. This protocol can also be modified for other genome-scale libraries, cell lines, and cytotoxins. For complete details on the use and execution of this protocol, please refer to Luo et al. (2022).
CRISPR 筛选是一种强大的工具，可用于鉴定包括病毒和细菌毒素在内的病原体的宿主因素。在这里，我们介绍了一种在 HeLa 细胞上进行全基因组 CRISPR 筛选的方案，用于鉴定与 TcdB4 毒素作用相关的宿主因素。我们详细描述了如何制备文库、设置筛选、获得基因序列以及分析结果。该方案也可以针对其他全基因组文库、细胞系和细胞毒素进行修改。有关此方案的使用和执行的完整详细信息，请参阅 Luo 等人（2022 年）。
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