CRISPR/Cas9系统在寄生虫基因组编辑中的应用研究进展

Research Progress of CRISPR/Cas9 System in Parasite Genome Editing

基因编辑是一种依赖于核酸内切酶对目标基因进行定向改造的现代生物学技术,可实现特定片段的加入、删除,以及特定碱基的插入、缺失、替换等。CRISPR/Cas9系统(clustered regularly interspaced short palindromic repeats/associated nuclease 9)是在细菌和古细菌中发现的一种用来抵御外源质粒和噬菌体入侵的获得性免疫系统,经改造后已成为一种新型的基因组编辑工具,因其具有操作简单、成本低、切割效率高等优点被广泛应用于多种动物、植物及微生物等的基因组编辑。应用CRISPR/Cas9系统在进行基因编辑的基础上可分析和验证基因功能、研究遗传育种及筛选药物靶点和疫苗分子等,以达到减弱病原微生物致病性、增强动植物抗病性、治疗遗传性疾病、病毒性感染及肿瘤等目的。寄生虫因其生活史复杂,与细菌和病毒相比基因组较大、结构较复杂,缺乏有效的研究工具等原因,使得相关研究进展缓慢,而CRISPR/Cas9基因编辑技术的出现为基因功能研究提供了新的技术手段。笔者对CRISPR/Cas9系统的发现及其在寄生虫基因组编辑中的应用研究进展进行综述,并对该系统的应用现状和发展进行总结和展望。

Gene editing is a modern biological technology that relies on endonuclease for directional modification of target genes, it can achieve the addition and deletion of specific fragments and the insertion, deletion and replacement of specific bases.The CRISPR/Cas9(clustered regularly interspaced short palindromic repeats/associated nuclease 9) system is an acquired immune system found in bacteria and archaea to resist the invasion of foreign plasmids and phages.After modification, it has become a novel genome editing tool.Because of its advantages of simple operation, low cost and high cutting efficiency, it is widely used in genome editing of various animals, plants and microorganisms.CRISPR/Cas9 system can be used to analyze and verify gene functions, study genetic breeding and screen drug targets and vaccine molecules on the basis of gene editing, so as to reduce the pathogenicity of pathogenic microorganisms, enhance the disease resistance of animals and plants, and treat genetic diseases, viral infections and tumors.Due to the complex life history of parasites, larger genome and more complex structure compared with bacteria and virus, and lack of effective research tools, relevant research progress is slow.However, the emergence of CRISPR/Cas9 gene editing technology provides a new technical means for gene function research.In this review, the discovery of CRISPR/Cas9 system and its application in parasite genome editing were reviewed, and the application status and development of CRISPR/Cas9 system were summarized and prospeced.