Suzuki Tatsuya, Saito Akatsuki
Institute for Advanced Co-Creation Studies, Research Institute for Microbial Diseases, Osaka University.
Department of Veterinary Medicine, Faculty of Agriculture, University of Miyazaki.
Nihon Yakurigaku Zasshi. 2022;157(2):134-138. doi: 10.1254/fpj.21072.
RNA viruses are responsible for several infectious diseases, including dengue fever, Zika fever, and COVID-19. Reverse genetics is a powerful tool to elucidate which domain or mutations in RNA viruses determine their pathogenicity and ability to evade antiviral drugs and host immune response. Previous reverse genetics systems for flaviviruses and coronaviruses have been technically challenging and time-consuming, thereby hampering the further understanding of events during viral evolution. A novel reverse genetics system-circular polymerase extension reaction (CPER)-has been developed to overcome this limitation. CPER is based on PCR-mediated assembly of DNA fragments that encode the whole genome of these viruses. CPER requires a relatively short time to introduce specific mutations into the viral genome of flaviviruses and SARS-CoV-2. In this review article, we explain the mode of action of this system and discuss the future direction of reverse genetics for RNA viruses.
RNA病毒引发了包括登革热、寨卡病毒病和新冠肺炎在内的多种传染病。反向遗传学是一种强大的工具，可用于阐明RNA病毒中的哪些结构域或突变决定了它们的致病性以及逃避抗病毒药物和宿主免疫反应的能力。以往用于黄病毒和冠状病毒的反向遗传学系统在技术上具有挑战性且耗时，从而阻碍了对病毒进化过程中事件的进一步了解。一种新型的反向遗传学系统——环化聚合酶延伸反应（CPER）——已被开发出来以克服这一限制。CPER基于PCR介导的对编码这些病毒全基因组的DNA片段进行组装。CPER在黄病毒和SARS-CoV-2的病毒基因组中引入特定突变所需时间相对较短。在这篇综述文章中，我们解释了该系统的作用模式，并讨论了RNA病毒反向遗传学的未来发展方向。
