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...

Descripción completa

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
Descripción
Sumario: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).

Ejemplares similares