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

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...

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Detalles Bibliográficos
Autores principales: Yang, Qi, Zhou, Yao, He, Liuqing, Zhang, Yuanyuan, Tao, Liang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9344021/
https://www.ncbi.nlm.nih.gov/pubmed/35928004
http://dx.doi.org/10.1016/j.xpro.2022.101595
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author Yang, Qi
Zhou, Yao
He, Liuqing
Zhang, Yuanyuan
Tao, Liang
author_facet Yang, Qi
Zhou, Yao
He, Liuqing
Zhang, Yuanyuan
Tao, Liang
author_sort Yang, Qi
collection PubMed
description 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).
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spelling pubmed-93440212022-08-03 Protocol for genome-wide CRISPR knockout screens of bacterial cytotoxins in HeLa cells Yang, Qi Zhou, Yao He, Liuqing Zhang, Yuanyuan Tao, Liang STAR Protoc Protocol 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). Elsevier 2022-07-31 /pmc/articles/PMC9344021/ /pubmed/35928004 http://dx.doi.org/10.1016/j.xpro.2022.101595 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Yang, Qi
Zhou, Yao
He, Liuqing
Zhang, Yuanyuan
Tao, Liang
Protocol for genome-wide CRISPR knockout screens of bacterial cytotoxins in HeLa cells
title Protocol for genome-wide CRISPR knockout screens of bacterial cytotoxins in HeLa cells
title_full Protocol for genome-wide CRISPR knockout screens of bacterial cytotoxins in HeLa cells
title_fullStr Protocol for genome-wide CRISPR knockout screens of bacterial cytotoxins in HeLa cells
title_full_unstemmed Protocol for genome-wide CRISPR knockout screens of bacterial cytotoxins in HeLa cells
title_short Protocol for genome-wide CRISPR knockout screens of bacterial cytotoxins in HeLa cells
title_sort protocol for genome-wide crispr knockout screens of bacterial cytotoxins in hela cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9344021/
https://www.ncbi.nlm.nih.gov/pubmed/35928004
http://dx.doi.org/10.1016/j.xpro.2022.101595
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