Todor Horia, Silvis Melanie R, Osadnik Hendrik, Gross Carol A
Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94158, USA.
Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94158, USA; Department of Cell and Tissue Biology, University of California, San Francisco, San Francisco, CA 94158, USA; California Institute of Quantitative Biology, University of California, San Francisco, San Francisco 94158 CA, USA.
Curr Opin Microbiol. 2021 Feb;59:102-109. doi: 10.1016/j.mib.2020.11.005. Epub 2020 Dec 4.
In this review we describe the application of CRISPR tools for functional genomics screens in bacteria, with a focus on the use of interference (CRISPRi) approaches. We review recent developments in CRISPRi titration, which has enabled essential gene functional screens, and genome-scale pooled CRISPRi screens. We summarize progress toward enabling CRISPRi screens in non-model and pathogenic bacteria, including the development of new dCas9 variants. Taking into account the current state of the field, we provide a forward-looking analysis of CRISPRi strategies for determining gene function in bacteria.
在本综述中，我们描述了CRISPR工具在细菌功能基因组学筛选中的应用，重点是干扰（CRISPRi）方法的使用。我们回顾了CRISPRi滴定法的最新进展，该方法已实现了必需基因功能筛选和全基因组规模的CRISPRi筛选。我们总结了在非模式细菌和致病细菌中进行CRISPRi筛选方面取得的进展，包括新型dCas9变体的开发。考虑到该领域的现状，我们对用于确定细菌基因功能的CRISPRi策略进行了前瞻性分析。
