Advances in utilizing the endogenous CRISPR-Cas system for genome editing of lactic acid bacteria

Abstract

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 Streptococcus pyogenes. 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.