Rapid identification of coronavirus replicase inhibitors using a selectable replicon RNA.
A previously unknown coronavirus (CoV) is the aetiological agent causing severe acute respiratory syndrome (SARS), for which an effective antiviral treatment is urgently needed. To enable the rapid and biosafe identification of coronavirus replicase inhibitors, we have generated a non-cytopathic, selectable replicon RNA (based on human CoV 229E) that can be stably maintained in eukaryotic cells. Most importantly, the replicon RNA mediates reporter gene expression as a marker for coronavirus replication. We have used a replicon RNA-containing cell line to test the inhibitory effect of several compounds that are currently being assessed for SARS treatment. Amongst those, interferon-alpha displayed the strongest inhibitory activity. Our results demonstrate that coronavirus replicon cell lines provide a versatile and safe assay for the identification of coronavirus replicase inhibitors. Once this technology is adapted to SARS-CoV replicon RNAs, it will allow high throughput screening for SARS-CoV replicase inhibitors without the need to grow infectious SARS-CoV.
Hertzig
Tobias
T
Institute of Virology and Immunology, University of Würzburg, Würzburg, Germany.
Scandella
Elke
E
Research Department, Cantonal Hospital St Gallen, CH-9007 St Gallen, Switzerland.
Schelle
Barbara
B
Institute of Virology and Immunology, University of Würzburg, Würzburg, Germany.
Ziebuhr
John
J
Institute of Virology and Immunology, University of Würzburg, Würzburg, Germany.
Siddell
Stuart G
SG
Department of Pathology and Microbiology, School of Medical Sciences, University of Bristol, Bristol, UK.
Ludewig
Burkhard
B
Research Department, Cantonal Hospital St Gallen, CH-9007 St Gallen, Switzerland.
Thiel
Volker
V
Research Department, Cantonal Hospital St Gallen, CH-9007 St Gallen, Switzerland.
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D012367:RNA, Viral / Q000502:physiology*; D012324:RNA-Dependent RNA Polymerase / Q000037:antagonists & inhibitors*; D012093:Replicon; D045473:Severe acute respiratory syndrome-related coronavirus / Q000187:drug effects* / Q000235:genetics*; D014779:Virus Replication / Q000187:drug effects*
D000998:Antiviral Agents; D012367:RNA, Viral; D012324:RNA-Dependent RNA Polymerase