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用于基因功能研究的细菌CRISPR筛选。

Bacterial CRISPR screens for gene function.

作者信息

Todor Horia, Silvis Melanie R, Osadnik Hendrik, Gross Carol A

机构信息

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.

出版信息

Curr Opin Microbiol. 2021 Feb;59:102-109. doi: 10.1016/j.mib.2020.11.005. Epub 2020 Dec 4.

Abstract

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.

摘要

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。

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