Department of Chemical and Pharmaceutical Biology, Groningen Research Institute of Pharmacy, University of Groningen, Groningen, The Netherlands.
Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Acadamy of Sciences, Guangzhou, China.
J Appl Microbiol. 2022 Oct;133(4):2280-2298. doi: 10.1111/jam.15704. Epub 2022 Jul 19.
Recently, the clustered regularly interspaced short palindromic repeats (CRISPR) system has been developed into a precise and efficient genome editing tool. Since its discovery as an adaptive immune system in prokaryotes, it has been applied in many different research fields including biotechnology and medical sciences. The high demand for rapid, highly efficient and versatile genetic tools to thrive in bacteria-based cell factories accelerates this process. This review mainly focuses on significant advancements of the CRISPR system in Bacillus subtilis, including the achievements in gene editing, and on problems still remaining. Next, we comprehensively summarize this genetic tool's up-to-date development and utilization in other Bacillus species, including B. licheniformis, B. methanolicus, B. anthracis, B. cereus, B. smithii and B. thuringiensis. Furthermore, we describe the current application of CRISPR tools in phages to increase Bacillus hosts' resistance to virulent phages and phage genetic modification. Finally, we suggest potential strategies to further improve this advanced technique and provide insights into future directions of CRISPR technologies for rendering Bacillus species cell factories more effective and more powerful.
最近，成簇规律间隔短回文重复（CRISPR）系统已发展成为一种精确且高效的基因组编辑工具。自其在原核生物中被发现作为适应性免疫系统以来，它已被应用于许多不同的研究领域，包括生物技术和医学科学。对在基于细菌的细胞工厂中快速、高效和多功能遗传工具的高需求加速了这一过程。本文主要综述了 CRISPR 系统在枯草芽孢杆菌中的重要进展，包括基因编辑方面的成就以及仍存在的问题。其次，我们全面总结了该遗传工具在其他芽孢杆菌属物种中的最新发展和利用，包括地衣芽孢杆菌、产甲烷芽孢杆菌、炭疽芽孢杆菌、解淀粉芽孢杆菌、苏云金芽孢杆菌和短小芽孢杆菌。此外，我们还描述了 CRISPR 工具在噬菌体中的应用，以提高芽孢杆菌宿主对毒性噬菌体和噬菌体遗传修饰的抗性。最后，我们提出了进一步改进该先进技术的潜在策略，并探讨了未来 CRISPR 技术在使芽孢杆菌属细胞工厂更有效和更强大方面的方向。
