Zhu Qing, Xu Chen, Zhang Shuwen, Xie Ning, Pang Xiaoyang, Lü Jiaping
Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Sheng Wu Gong Cheng Xue Bao. 2022 Jul 25;38(7):2447-2458. doi: 10.13345/j.cjb.210958.
The CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated proteins) system is an efficient and precise gene editing tool. The development of this technology has promoted genome editing into a rapid development stage. The most widely used Cas9 protein is SpyCas9 from . As a "gene scissors", the SpyCas9 protein is widely used in eukaryotes including mammals and plants. However, the application of this protein in some lactic acid bacteria (LAB) is still hampered by many factors. It has been identified that LAB genomes possess many types of CRISPR system and are rich in uncharacterized Cas proteins. Genome editing of LAB is possible by repurposing the endogenous CRISPR-Cas systems in LAB combined with exogenous single guide RNA (sgRNA) and homologous recombination template. This method employs its endogenous CRISPR-Cas system for gene editing, which has the advantages including easiness for transformation due to the relatively small targeting vector, and no concern about the toxicity of heterologous Cas9 to host cells. Compared to CRISPR-SpyCas9, the endogenous CRISPR-Cas system is more suitable for genome editing of LAB, and it may become the main genome editing tool for some LAB in the future. This article summarizes the advances in this field.
CRISPR（成簇规律间隔短回文重复序列）-Cas（CRISPR相关蛋白）系统是一种高效且精确的基因编辑工具。该技术的发展推动了基因组编辑进入快速发展阶段。应用最为广泛的Cas9蛋白是来自[具体来源未给出]的SpyCas9。作为一把“基因剪刀”，SpyCas9蛋白在包括哺乳动物和植物在内的真核生物中被广泛使用。然而，这种蛋白在一些乳酸菌（LAB）中的应用仍受到诸多因素的阻碍。已确定LAB基因组拥有多种类型的CRISPR系统，并且富含未表征的Cas蛋白。通过将LAB中的内源性CRISPR-Cas系统与外源性单导向RNA（sgRNA）和同源重组模板相结合，LAB的基因组编辑成为可能。该方法利用其内源CRISPR-Cas系统进行基因编辑，具有诸多优点，包括由于靶向载体相对较小而易于转化，以及无需担心异源Cas9对宿主细胞的毒性。与CRISPR-SpyCas9相比，内源性CRISPR-Cas系统更适合LAB的基因组编辑，并且未来它可能会成为一些LAB的主要基因组编辑工具。本文总结了该领域的进展。
