Inhaled nebulized interferon (IFN)-α and IFN-β have been shown to be effective in the management of coronavirus disease 2019 (COVID-19). We aimed to construct a virus-free rapid detection system for high-throughput screening of IFN-like compounds that induce viral RNA degradation and suppress the replication of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We prepared a SARS-CoV-2 subreplicon RNA expression vector which contained the SARS-CoV-2 5′-UTR, the partial sequence of ORF1a, luciferase, nucleocapsid, ORF10, and 3′-UTR under the control of the cytomegalovirus promoter. The expression vector was transfected into Calu-3 cells and treated with IFN-α and the IFNAR2 agonist CDM-3008 (RO8191) for 3 days. SARS-CoV-2 subreplicon RNA degradation was subsequently evaluated based on luciferase levels. IFN-α and CDM-3008 suppressed SARS-CoV-2 subreplicon RNA in a dose-dependent manner, with IC50 values of 193 IU/mL and 2.54 μM, respectively. HeLa cells stably expressing SARS-CoV-2 subreplicon RNA were prepared and treated with the IFN-α and pan-JAK inhibitor Pyridone 6 or siRNA-targeting ISG20. IFN-α activity was canceled with Pyridone 6. The knockdown of ISG20 partially canceled IFN-α activity. Collectively, we constructed a virus-free rapid detection system to measure SARS-CoV-2 RNA suppression. Our data suggest that the SARS-CoV-2 subreplicon RNA was degraded by IFN-α-induced ISG20 exonuclease activity.
【저자키워드】 COVID-19, interferon, replicon, interferon-stimulated genes, ISG20, CDM-3008, 【초록키워드】 coronavirus disease, SARS-CoV-2, coronavirus, IC50, RNA, Replication, nucleocapsid, management, SARS-CoV-2 RNA, Viral RNA, Degradation, Calu-3 cell, inhibitor, ORF10, exonuclease, IFNAR2, RNA degradation, IFN-α, acute respiratory syndrome, partial sequence, IFN-β, dose-dependent manner, Compound, nebulized, ORF1a, knockdown, promoter, RNA expression, 5′-UTR, expression vector, 3′-UTR, effective, detection system, shown, evaluated, treated, induce, suppressed, suppress, expressing, HeLa cell, Pyridone 6, the SARS-CoV-2, 【제목키워드】 SARS-CoV-2, detection, RNA, Rapid, induced, System,