CRISPR/Cas9系统在疾病研究和治疗中的应用

CRISPR/Cas9 technology in disease research and therapy: a review

摘要:

基因组编辑技术 (Genome editing technology) 是一种通过人工手段在基因组水平对DNA序列进行改造的遗传操作技术，包括特定DNA片段的插入、敲除、替换和点突变。其中，依赖核酸酶的基因组编辑技术的基本原理是在基因组的特定位置产生双链DNA断裂 (Double-stranded break，DSB) 后通过非同源末端连接 (Non-homologous end joining，NHEJ) 或同源重组 (Homologous recombination，HR) 的方式进行修复。随着对核酸酶更深入的研究，基因组编辑技术也得到了快速发展，其中最常使用的核酸酶主要包括巨型核酸酶、锌指核酸酶、转录激活因子样效应物核酸酶以及成簇的规律间隔的短回文重复序列相关蛋白 (Clustered regularly interspaced short palindromic repeats (CRISPR)-associated proteins (Cas)。文中在介绍上述基因组编辑技术的发展及作用原理的基础上，主要综述了CRISPR/Cas9系统在基因功能鉴定、疾病模型建立、基因治疗和免疫治疗等应用领域的研究进展，并对其未来发展进行了展望。

Abstract:

Genome editing is a genetic manipulation technique that can modify DNA sequences at the genome level, including insertion, knockout, replacement and point mutation of specific DNA fragments. The ultimate principle of genome editing technology relying on engineered nucleases is to generate double-stranded DNA breaks at specific locations in genome and then repair them through non-homologous end joining or homologous recombination. With the intensive study of these nucleases, genome editing technology develops rapidly. The most used nucleases include meganucleases, zinc finger nucleases, transcription activator-like effector nucleases, and clustered regularly interspaced short palindromic repeats associated Cas proteins. Based on introducing the development and principles of above mentioned genome editing technologies, we review the research progress of CRISPR/Cas9 system in the application fields of identification of gene function, establishment of disease model, gene therapy, immunotherapy and its prospect.