Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Microbiol Mol Biol Rev. 2024 Jun 27;88(2):e0017022. doi: 10.1128/mmbr.00170-22. Epub 2024 May 29.
SUMMARYFunctional genomics is the use of systematic gene perturbation approaches to determine the contributions of genes under conditions of interest. Although functional genomic strategies have been used in bacteria for decades, recent studies have taken advantage of CRISPR (clustered regularly interspaced short palindromic repeats) technologies, such as CRISPRi (CRISPR interference), that are capable of precisely modulating expression of all genes in the genome. Here, we discuss and review the use of CRISPRi and related technologies for bacterial functional genomics. We discuss the strengths and weaknesses of CRISPRi as well as design considerations for CRISPRi genetic screens. We also review examples of how CRISPRi screens have defined relevant genetic targets for medical and industrial applications. Finally, we outline a few of the many possible directions that could be pursued using CRISPR-based functional genomics in bacteria. Our view is that the most exciting screens and discoveries are yet to come.
摘要功能基因组学是利用系统的基因干扰方法来确定在感兴趣的条件下基因的贡献。尽管功能基因组学策略在细菌中已经使用了几十年，但最近的研究利用了 CRISPR（成簇规律间隔短回文重复）技术，如 CRISPRi（CRISPR 干扰），能够精确地调节基因组中所有基因的表达。在这里，我们讨论和回顾了 CRISPRi 和相关技术在细菌功能基因组学中的应用。我们讨论了 CRISPRi 的优缺点以及 CRISPRi 遗传筛选的设计考虑因素。我们还回顾了 CRISPRi 筛选如何为医疗和工业应用定义相关遗传靶标的例子。最后，我们概述了使用基于 CRISPR 的功能基因组学在细菌中可能追求的许多方向。我们的观点是，最令人兴奋的筛选和发现还在后面。
