Visualization of Positive and Negative Sense Viral RNA for Probing the Mechanism of Direct-Acting Antivirals against Hepatitis C Virus
1
CS Bond Life Sciences Center, University of Missouri, Columbia, MO 65201, USA
2
Department of Molecular Microbiology & Immunology, University of Missouri School of Medicine, Columbia, MO 65212, USA
3
Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, USA
4
Department of Veterinary Pathobiology, University of Missouri, Columbia, MO 65211, USA
5
Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA
6
Department of Biochemistry, University of Missouri, Columbia, MO 65201, USA
*
Author to whom correspondence should be addressed.
†
Current address: Department of Molecular Microbiology, Washington University School of Medicine, Saint Louis, MO 63110, USA.
Viruses 2019, 11(11), 1039; https://doi.org/10.3390/v11111039
Received: 11 October 2019 / Revised: 3 November 2019 / Accepted: 7 November 2019 / Published: 8 November 2019
(This article belongs to the Section Animal Viruses)
RNA viruses are highly successful pathogens and are the causative agents for many important diseases. To fully understand the replication of these viruses it is necessary to address the roles of both positive-strand RNA ((+)RNA) and negative-strand RNA ((−)RNA), and their interplay with viral and host proteins. Here we used branched DNA (bDNA) fluorescence in situ hybridization (FISH) to stain both the abundant (+)RNA and the far less abundant (−)RNA in both hepatitis C virus (HCV)- and Zika virus-infected cells, and combined these analyses with visualization of viral proteins through confocal imaging. We were able to phenotypically examine HCV-infected cells in the presence of uninfected cells and revealed the effect of direct-acting antivirals on HCV (+)RNA, (−)RNA, and protein, within hours of commencing treatment. Herein, we demonstrate that bDNA FISH is a powerful tool for the study of RNA viruses that can provide insights into drug efficacy and mechanism of action.
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Keywords:
hepatitis C virus; Zika virus; positive-sense; negative-sense; direct acting antivirals; RNA FISH
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited
MDPI and ACS Style
Liu, D.; Tedbury, P.R.; Lan, S.; Huber, A.D.; Puray-Chavez, M.N.; Ji, J.; Michailidis, E.; Saeed, M.; Ndongwe, T.P.; Bassit, L.C.; Schinazi, R.F.; Ralston, R.; Rice, C.M.; Sarafianos, S.G. Visualization of Positive and Negative Sense Viral RNA for Probing the Mechanism of Direct-Acting Antivirals against Hepatitis C Virus. Viruses 2019, 11, 1039.
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