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Protocol for genome-wide CRISPR knockout screens of bacterial cytotoxins in HeLa cells.

ABSTRACT: 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 Clostridioides difficile 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).

SUBMITTER: Yang Q

PROVIDER: S-EPMC9344021 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature

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Protocol for genome-wide CRISPR knockout screens of bacterial cytotoxins in HeLa cells.

Yang Qi Q Zhou Yao Y He Liuqing L Zhang Yuanyuan Y Tao Liang L

STAR protocols 20220731 3

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 <i>Clostridioides difficile</i> 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 c ...[more]

PMID: 35928004

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