Zhang Xianwen, Liu Yang, Liu Jianying, Bailey Adam L, Plante Kenneth S, Plante Jessica A, Zou Jing, Xia Hongjie, Bopp Nathen, Aguilar Patricia, Ren Ping, Menachery Vineet D, Diamond Michael S, Weaver Scott C, Xie Xuping, Shi Pei-Yong
bioRxiv. 2021 Jan 19:2021.01.16.426970. doi: 10.1101/2021.01.16.426970.
The biosafety level-3 (BSL-3) requirement to culture severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a bottleneck for research and countermeasure development. Here we report a -complementation system that produces single-round infectious SARS-CoV-2 that recapitulates authentic viral replication. We demonstrate that the single-round infectious SARS-CoV-2 can be used at BSL-2 laboratories for high-throughput neutralization and antiviral testing. The -complementation system consists of two components: a genomic viral RNA containing a deletion of ORF3 and envelope gene, and a producer cell line expressing the two deleted genes. complementation of the two components generates virions that can infect naive cells for only one round, but does not produce wild-type SARS-CoV-2. Hamsters and K18-hACE2 transgenic mice inoculated with the complementation-derived virions exhibited no detectable disease, even after intracranial inoculation with the highest possible dose. The results suggest that the -complementation platform can be safely used at BSL-2 laboratories for research and countermeasure development.
对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）进行培养的生物安全3级（BSL-3）要求是研究和对策开发的一个瓶颈。在此，我们报告一种互补系统，该系统可产生单轮感染性SARS-CoV-2，其概括了真实的病毒复制过程。我们证明，单轮感染性SARS-CoV-2可在BSL-2实验室用于高通量中和及抗病毒测试。该互补系统由两个组件组成：一个基因组病毒RNA，其缺失了开放阅读框3（ORF3）和包膜基因；一个表达这两个缺失基因的生产细胞系。这两个组件的互补产生的病毒粒子只能感染未感染细胞一轮，但不会产生野生型SARS-CoV-2。接种了互补衍生病毒粒子的仓鼠和K18-hACE2转基因小鼠，即使在以最高可能剂量进行颅内接种后，也未表现出可检测到的疾病。结果表明，该互补平台可在BSL-2实验室安全用于研究和对策开发。
