Abstract
The biosafety level 3 (BSL-3) requirement to culture severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a bottleneck for research. Here, we report a trans-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 trans-complementation system consists of two components: a genomic viral RNA containing ORF3 and envelope gene deletions, as well as mutated transcriptional regulator sequences, and a producer cell line expressing the two deleted genes. Trans-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. Thus, the trans-complementation platform can be safely used at BSL-2 laboratories for research and countermeasure development.
Keywords: COVID-19; SARS-CoV-2; antiviral; coronavirus; diagnosis; vaccine.
【저자키워드】 COVID-19, SARS-CoV-2, Vaccine, coronavirus, Antiviral, Diagnosis, 【초록키워드】 Vaccine, coronavirus, Antiviral, hamsters, neutralization, severe acute respiratory syndrome Coronavirus, Viral, Culture, viral replication, Research, Viral RNA, hamster, respiratory, genomic, disease, platform, K18-hACE2 transgenic mice, dose, biosafety level, level 3, gene deletions, ORF3, acute respiratory syndrome, acute respiratory syndrome coronavirus, acute respiratory syndrome coronavirus 2, wild-type SARS-CoV-2, trans, BSL-2 laboratory, component, cell line, virion, virions, deletions, naive cells, envelope gene, transcriptional regulator, infect, Complementation, Genes, Cell, Biosafety Level 3, BSL-3, highest, inoculated, detectable, generate, exhibited, can be used, mutated, expressing, deleted, 【제목키워드】 viral replication, virulence,