An RNA polymerase II-driven Ebola virus minigenome system as an advanced tool for antiviral drug screening

Emily V. Nelson, Jennifer R. Pacheco, Adam J. Hume, Tessa N. Cressey, Laure R. Deflubé, John B. Ruedas, John H. Connor, Hideki Ebihara, Elke Mühlberger

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)21-27
Number of pages7
JournalAntiviral Research
Volume146
DOIs
StatePublished - Oct 1 2017

Fingerprint

Ebolavirus
Preclinical Drug Evaluations
RNA Polymerase II
Antiviral Agents
Ebola Hemorrhagic Fever
Virus Replication
Research
Vaccines
Public Health
RNA
Costs and Cost Analysis

Keywords

  • Antiviral drug screening
  • Ebola virus
  • Filoviruses
  • Minigenome system
  • RNA polymerase II
  • T7 RNA polymerase

ASJC Scopus subject areas

  • Pharmacology
  • Virology

Cite this

Nelson, E. V., Pacheco, J. R., Hume, A. J., Cressey, T. N., Deflubé, L. R., Ruedas, J. B., ... Mühlberger, E. (2017). An RNA polymerase II-driven Ebola virus minigenome system as an advanced tool for antiviral drug screening. Antiviral Research, 146, 21-27. https://doi.org/10.1016/j.antiviral.2017.08.005

An RNA polymerase II-driven Ebola virus minigenome system as an advanced tool for antiviral drug screening. / Nelson, Emily V.; Pacheco, Jennifer R.; Hume, Adam J.; Cressey, Tessa N.; Deflubé, Laure R.; Ruedas, John B.; Connor, John H.; Ebihara, Hideki; Mühlberger, Elke.

In: Antiviral Research, Vol. 146, 01.10.2017, p. 21-27.

Research output: Contribution to journalArticle

Nelson, EV, Pacheco, JR, Hume, AJ, Cressey, TN, Deflubé, LR, Ruedas, JB, Connor, JH, Ebihara, H & Mühlberger, E 2017, 'An RNA polymerase II-driven Ebola virus minigenome system as an advanced tool for antiviral drug screening', Antiviral Research, vol. 146, pp. 21-27. https://doi.org/10.1016/j.antiviral.2017.08.005
Nelson, Emily V. ; Pacheco, Jennifer R. ; Hume, Adam J. ; Cressey, Tessa N. ; Deflubé, Laure R. ; Ruedas, John B. ; Connor, John H. ; Ebihara, Hideki ; Mühlberger, Elke. / An RNA polymerase II-driven Ebola virus minigenome system as an advanced tool for antiviral drug screening. In: Antiviral Research. 2017 ; Vol. 146. pp. 21-27.
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