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10.3390/ijms222111641

http://scihub22266oqcxt.onion/10.3390/ijms222111641
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34769072!8583800!34769072
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suck abstract from ncbi


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pmid34769072      Int+J+Mol+Sci 2021 ; 22 (21): ä
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  • Establishment of a Rapid Detection System for ISG20-Dependent SARS-CoV-2 Subreplicon RNA Degradation Induced by Interferon-alpha #MMPMID34769072
  • Furutani Y; Toguchi M; Higuchi S; Yanaka K; Gailhouste L; Qin XY; Masaki T; Ochi S; Matsuura T
  • Int J Mol Sci 2021[Oct]; 22 (21): ä PMID34769072show ga
  • Inhaled nebulized interferon (IFN)-alpha and IFN-beta 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-alpha and the IFNAR2 agonist CDM-3008 (RO8191) for 3 days. SARS-CoV-2 subreplicon RNA degradation was subsequently evaluated based on luciferase levels. IFN-alpha and CDM-3008 suppressed SARS-CoV-2 subreplicon RNA in a dose-dependent manner, with IC50 values of 193 IU/mL and 2.54 muM, respectively. HeLa cells stably expressing SARS-CoV-2 subreplicon RNA were prepared and treated with the IFN-alpha and pan-JAK inhibitor Pyridone 6 or siRNA-targeting ISG20. IFN-alpha activity was canceled with Pyridone 6. The knockdown of ISG20 partially canceled IFN-alpha 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-alpha-induced ISG20 exonuclease activity.
  • |Antiviral Agents/*pharmacology[MESH]
  • |Cell Line, Tumor[MESH]
  • |Dose-Response Relationship, Drug[MESH]
  • |Drug Evaluation, Preclinical/*methods[MESH]
  • |Exoribonucleases/genetics[MESH]
  • |Genetic Vectors[MESH]
  • |HeLa Cells[MESH]
  • |Humans[MESH]
  • |Interferon-alpha/administration & dosage/*pharmacology[MESH]
  • |Luciferases/genetics/metabolism[MESH]
  • |Naphthyridines/administration & dosage/pharmacology[MESH]
  • |Oxadiazoles/administration & dosage/pharmacology[MESH]
  • |RNA, Viral/drug effects/*metabolism[MESH]
  • |Replicon[MESH]


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