An RNA polymerase II-driven Ebola virus minigenome system as an advanced tool for antiviral drug screening
Autor: | Elke Mühlberger, Emily V. Nelson, John H. Connor, Laure R. Deflubé, Adam J. Hume, Hideki Ebihara, Jennifer R. Pacheco, Tessa N. Cressey, John B. Ruedas |
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Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Transcription Genetic medicine.drug_class HSP72 Heat-Shock Proteins RNA polymerase II Genome Viral Microbial Sensitivity Tests Biology Virus Replication medicine.disease_cause Antiviral Agents Article Viral Proteins 03 medical and health sciences Transcription (biology) Virology Biosafety level medicine Animals Humans T7 RNA polymerase Antiviral screening Pharmacology Ebola virus RNA DNA-Directed RNA Polymerases Hemorrhagic Fever Ebola Ebolavirus High-Throughput Screening Assays 030104 developmental biology biology.protein RNA Viral RNA Polymerase II Antiviral drug medicine.drug |
Zdroj: | Antiviral Research. 146:21-27 |
ISSN: | 0166-3542 |
Popis: | Ebola virus (EBOV) causes a severe disease in humans with the potential for significant international public health consequences. Currently, treatments are limited to experimental vaccines and therapeutics. Therefore, research into prophylaxis and antiviral strategies to combat EBOV infections is of utmost importance. The requirement for high containment laboratories to study EBOV infection is a limiting factor for conducting EBOV research. To overcome this issue, minigenome systems have been used as valuable tools to study EBOV replication and transcription mechanisms and to screen for antiviral compounds at biosafety level 2. The most commonly used EBOV minigenome system relies on the ectopic expression of the T7 RNA polymerase (T7), which can be limiting for certain cell types. We have established an improved EBOV minigenome system that utilizes endogenous RNA polymerase II (pol II) as a driver for the synthesis of minigenome RNA. We show here that this system is as efficient as the T7-based minigenome system, but works in a wider range of cell types, including biologically relevant cell types such as bat cells. Importantly, we were also able to adapt this system to a reliable and cost-effective 96-well format antiviral screening assay with a Z-factor of 0.74, indicative of a robust assay. Using this format, we identified JG40, an inhibitor of Hsp70, as an inhibitor of EBOV replication, highlighting the potential for this system as a tool for antiviral drug screening. In summary, this updated EBOV minigenome system provides a convenient and effective means of advancing the field of EBOV research. |
Databáze: | OpenAIRE |
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