基于CRISPR-Cas系统的多重耐药菌防治技术研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2024-0872

摘要/Abstract

摘要：

动物源细菌耐药性影响动物养殖安全，同时对人类公共卫生和食品安全产生重要威胁。抗生素的滥用加剧了耐药细菌的传播，而新型抗菌药物研发日益困难，动物源细菌耐药污染已成为全球范围内的公共危机，一旦耐药菌从动物向人类扩散，将极大地威胁人类健康，亟须新的方法和策略应对细菌耐药。CRISPR-Cas（clustered regularly interspaced short palindromic repeats, CRISPR-associated）是第三代“基因组定点编辑技术”，该技术能够靶向剪切外源性核酸，保护微生物遗传物质遗传稳定性。与传统的多重耐药菌防治策略相比，CRISPR-Cas系统具备独特的DNA序列的靶向性和灵敏度，通过精准、简便和高效的基因编辑技术，与核酸扩增技术、比色技术等相结合，可以提高灵敏度和检测时效性等性能指标。本文介绍了CRISPR-Cas系统的由来、系统分类、基因编辑的作用机理，在此基础之上，聚焦于该系统在多重耐药菌防治领域的研究进展和应用，在耐药致病菌消除、耐药基因消除以及致病菌诊断检测方面给出案例分析以及目前存在的挑战。总的来说，基于CRISPR-Cas系统的序列特异性抗菌剂能够降低细菌多重耐药性，结合核酸扩增技术和实时监控设备提升检测的精准性和效率，为动物源细菌耐药防控和监测研究提供了新思路。

关键词: CRISPR-Cas系统, 基因编辑, 多重耐药菌, 耐药基因, 防治技术

Abstract:

Antimicrobial resistance of animal-derived bacteria affects the safety of animal breeding and causes an important threat to human public health and food safety. The abuse of antibiotics intensifies the spread of drug-resistant bacteria, and the research and development of new antibacterial drugs is becoming increasingly difficult. Drug-resistant bacteria contamination of animal-derived bacteria has become a global public crisis. Once drug-resistant bacteria spread from animals to humans, it will greatly threaten human health, and we urgently need new methods and strategies to deal with bacterial resistance. CRISPR-Cas (clustered regularly interspaced short palindromic repeats CRISPR-associated) is the third generation of "genome point editing technology". This technology can target the cutting of exogenous DNA and protect the genetic stability of microbial genetic material. Compared with traditional multi-drug-resistant bacteria control strategies, CRISPR-Cas system has a unique DNA sequence targeting and sensitivity. The sensitivity and detection timeliness and other performance indicators can be improved through accurate, simple and efficient gene editing technology, combined with nucleic acid amplification technology, colorimetric technology. This paper introduces the origin, systematic classification, and mechanism of gene editing of CRISPR-Cas system. Based on this brief introduction, it focuses on the research progress and application of the system in the field of multi-drug-resistant bacteria prevention and control, and have case studies and current challenges in the elimination of drug-resistant pathogens, the elimination of drug-resistant genes, and the diagnosis and detection of pathogens. In general, sequence-specific antimicrobials based on CRISPR-Cas system may reduce bacterial multidrug resistance, and the combination of nucleic acid amplification technology and real-time monitoring equipment may improve the accuracy and efficiency of detection, providing a new idea for the prevention and control of drug resistance and monitoring of animal-derived bacteria.

Key words: CRISPR-Cas system, gene editing, multidrug resistant bacteria, antibiotic resistance genes, control technology

周倩, 唐梦君, 张小燕, 陆俊贤, 唐修君, 杨星星, 高玉时. 基于CRISPR-Cas系统的多重耐药菌防治技术研究进展[J]. 生物技术通报, 2025, 41(5): 42-51.

ZHOU Qian, TANG Meng-jun, ZHANG Xiao-yan, LU Jun-xian, TANG Xiu-jun, YANG Xing-xing, GAO Yu-shi. Research Progress in the Control of Multidrug Resistant Bacteria Based on in CRISPR-Cas System[J]. Biotechnology Bulletin, 2025, 41(5): 42-51.

图/表 4

图1 CRISPR结构图

Fig. 1 Structure of CRISPR

表1 CRISPR分型与Cas蛋白功能

Table 1 CRISPR types and the functions of Cas protein

类型 Type		适配 Adaption	表达 Expression	干扰 Interference	辅助 Ancillary
Class 1	Ⅰ type	Cas1-Cas2	Cas6	Cas7-Cas5-Cas8-Cas3	Cas4
	Ⅲ type	Cas1-Cas2	Cas6
	Ⅳ type			Cas7-Cas5
Class 2	Ⅱ type	Cas1-Cas2	RNase Ⅲ	Cas9	Cas4
	V type	Cas1-Cas2		Cas12	Cas4
	Ⅵ type	Cas1-Cas2		Cas13

图2 CRISPR-Cas系统作用机理

Fig. 2 Action mechanism of CRISPR-Cas system

图3 CRISPR-Cas抑菌递送的图形概念模型

Fig. 3 Graphical concepts model of CRISPR-Cas for antibacterial delivery

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