人冠状病毒传染性克隆及其应用研究进展

IF 3.5 Q1 PUBLIC, ENVIRONMENTAL & OCCUPATIONAL HEALTH

Biosafety and Health Pub Date : 2025-02-01 DOI:10.1016/j.bsheal.2025.01.003

Guanya Liu , Ruixiao Tan , Yiyi Wu , Mengwei Wang , Baoying Huang , Wenjie Tan

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引用次数: 0

摘要

冠状病毒可以感染人类、哺乳动物和鸟类，导致呼吸道、胃肠道和神经系统疾病。这些病毒是重要的人畜共患病原体，已知有九种类型能够感染人类。冠状病毒基因组约30 kb，是已知最大的核糖核酸（RNA）病毒基因组，其复杂性使得组装和操作既耗时又费力。反向遗传系统被广泛用于设计重组病毒，这些病毒可以适应生物安全水平2 (BSL-2)，用于研究病毒基因功能、复制、发病机制、疫苗和治疗方法。传染性克隆是病毒逆向遗传不可缺少的工具，它可以通过DNA重组技术恢复各种病毒。构建人类冠状病毒（HCoV）传染性克隆的各种技术已经发展起来，包括痘苗病毒载体法、体外结扎法、细菌人工染色体系统、酵母人工染色体系统、环状聚合酶延伸反应以及最近报道的传染性亚基因组扩增子技术。本文综述了构建人类冠状病毒传染性克隆的各种技术及其应用现状。

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Advances in the development of infectious clones of human coronaviruses and related applications

Coronaviruses can infect humans, mammals, and birds, leading to respiratory, gastrointestinal, and neurological diseases. These viruses are significant zoonotic pathogens with nine known types capable of infecting humans. The coronavirus genome, approximately 30 kb in size, is the largest known ribonucleic acid (RNA) virus genome, and its complexity makes assembly and manipulation time-consuming and labor-intensive. Reverse genetic systems are widely used to engineer recombinant viruses that can be adapted at Biosafety Level 2 (BSL-2) for studying viral gene function, replication, pathogenesis, vaccines, and therapeutics. The infectious clones, which enabled the recovery of various viruses after DNA recombinant technology, were indispensable tools for the reverse genetics of viruses. Various techniques for constructing infectious clones of human coronaviruses (HCoV) have been developed, encompassing methods such as vaccinia virus vectors method, in vitro ligation, bacterial artificial chromosome systems, yeast artificial chromosome systems, circular polymerase extension reaction, and the recently reported infectious sub-genomic amplicons technology. This review summarizes the status of various techniques for constructing infectious clones of human coronaviruses and related applications.

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