Ⅱ类CRISPR/Cas系统及其在细菌合成生物学中的应用

doi:10.11843/j.issn.0366-6964.2025.04.012

Abstract

Abstract:

The CRISPR/Cas system is an adaptive immune system widely present in bacteria and archaea. It targets and cleaves specific nucleic acid sequences using RNA-guided nucleases to defend against foreign nucleic acid invasion. Based on the composition of effector protein complexes, the CRISPR/Cas system can be divided into two main classes: Class I systems utilize effector complexes composed of multiple Cas proteins, acting together with crRNA to exert target cleavage function, while Class II systems consist of single, multi-domain protein effector modules. Among them, the single-component effector protein-mediated Class II CRISPR gene editing technology is relatively straightforward and has been widely applied in synthetic biology fields such as bacterial gene expression regulation, genetic modification, metabolic pathway optimization, and pathogenic microorganism detection. Based on the differences in the Cas effector protein nuclease domains, Class II systems can further be categorized into subtypes II, V, and VI, each exhibiting differences in their applications in bacterial synthetic biology. This article reviews the immunological mechanisms, classification, and characteristics of Class II CRISPR-Cas systems, as well as their current applications and the latest developments in industrial bacteria, aiming to provide a reference for future research in bacterial synthetic biology to select, optimize, and explore more CRISPR/Cas systems.

Key words: CRISPR/Cas system, bacteria, gene editing, synthetic biology, effector protein

CLC Number:

LI Xiaohan, LI Guiping, HUO Caiyun, ZHANG Qilong, SUN Yingjian, SUN Huiling. Class II CRISPR/Cas Systems and Their Applications in Bacterial Synthetic Biology[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(4): 1608-1620.

Figures/Tables 6

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类型 Type	Ⅱ型 Type Ⅱ	Ⅴ型 Type Ⅴ	Ⅵ型 Type Ⅵ
标志性效应蛋白 Signature effector protein	Cas9，II-A	Cas12a，V-A	Cas13a，VI-A
发源菌 Bacteria of origin	化脓性链球菌	新凶手弗朗西斯菌氨基酸球菌	纤毛菌瘤胃球菌
		氨基酸球菌	瘤胃球菌
		毛螺菌	普雷沃菌
		摩拉菌	李斯特菌
CRISPR-RNA	sgRNA，100 nt	crRNA，42~44 nt	crRNA，52~66 nt
PAM序列	G富集	T富集	无
PAM sequence	5′-NGG-3′	5′-TT(T)N-3′	PFS序列
核酸酶结构域Nuclease domain	HNH、RuvC	RuvC	HEPN
切割方式Cutting mode	dsDNA，平末端	dsDNA，黏末端	ssRNA
RNA酶活性RNAse activity	无	有	有

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Class II CRISPR/Cas Systems and Their Applications in Bacterial Synthetic Biology

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