Department of Chemical Engineering, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai, Osaka, 599-8531, Japan.
World J Microbiol Biotechnol. 2019 Jul 6;35(7):111. doi: 10.1007/s11274-019-2688-8.
Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) immune systems in bacteria have been used as tools for genome engineering. Thus far, the CRISPR-Cas system has been used in various yeast, bacterial, and mammalian cells. Saccharomyces cerevisiae is a nonpathogenic yeast, classified under "generally recognized as safe", and has long been used to produce consumables such as alcohol or bread. Additionally, recombinant cells of S. cerevisiae have been constructed and used to produce various bio-based chemicals. Some types of CRISPR-Cas system for genetic manipulation have been constructed during the early developmental stages of the CRISPR-Cas system and have been mainly used for gene knock-in and knock-out manipulations. Thereafter, these systems have been used for various novel purposes such as metabolic engineering and tolerance engineering. In this review, we have summarized different aspects of the CRISPR-Cas in the yeast S. cerevisiae, from its basic principles to various applications. This review describes the CRISPR system in S. cerevisiae based on the differences in its origin and efficiency followed by its basic applications; for example, its involvement in gene knock-in and knock-out has been outlined. Finally, advanced applications of the CRISPR system in the bioproduction of useful chemicals have been summarized.
细菌中的成簇规律间隔短回文重复 (CRISPR) 和 CRISPR 相关 (Cas) 免疫系统已被用作基因组工程的工具。迄今为止,CRISPR-Cas 系统已在各种酵母、细菌和哺乳动物细胞中得到应用。酿酒酵母是一种非致病性酵母,被归类为“普遍认为安全”,长期以来一直用于生产酒精或面包等消费品。此外,还构建了重组酿酒酵母细胞,并用于生产各种基于生物的化学品。在 CRISPR-Cas 系统的早期发展阶段构建了一些用于遗传操作的 CRISPR-Cas 系统类型,主要用于基因敲入和敲除操作。此后,这些系统已被用于各种新的用途,如代谢工程和耐受工程。在这篇综述中,我们总结了酿酒酵母 S. cerevisiae 中 CRISPR-Cas 的不同方面,从其基本原理到各种应用。本综述根据其起源和效率的差异描述了酿酒酵母中的 CRISPR 系统,例如,概述了其在基因敲入和敲除中的作用。最后,总结了 CRISPR 系统在有用化学品生物生产中的高级应用。