PMID- 31717338 OWN - NLM STAT- MEDLINE DCOM- 20200929 LR - 20200929 IS - 1999-4915 (Electronic) IS - 1999-4915 (Linking) VI - 11 IP - 11 DP - 2019 Nov 8 TI - Visualization of Positive and Negative Sense Viral RNA for Probing the Mechanism of Direct-Acting Antivirals against Hepatitis C Virus. LID - 10.3390/v11111039 [doi] LID - 1039 AB - 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. FAU - Liu, Dandan AU - Liu D AD - CS Bond Life Sciences Center, University of Missouri, Columbia, MO 65201, USA. AD - Department of Molecular Microbiology & Immunology, University of Missouri School of Medicine, Columbia, MO 65212, USA. FAU - Tedbury, Philip R AU - Tedbury PR AUID- ORCID: 0000-0001-8151-4967 AD - CS Bond Life Sciences Center, University of Missouri, Columbia, MO 65201, USA. AD - Department of Molecular Microbiology & Immunology, University of Missouri School of Medicine, Columbia, MO 65212, USA. AD - Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, USA. FAU - Lan, Shuiyun AU - Lan S AD - Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, USA. FAU - Huber, Andrew D AU - Huber AD AUID- ORCID: 0000-0002-3790-8368 AD - CS Bond Life Sciences Center, University of Missouri, Columbia, MO 65201, USA. AD - Department of Veterinary Pathobiology, University of Missouri, Columbia, MO 65211, USA. FAU - Puray-Chavez, Maritza N AU - Puray-Chavez MN AD - CS Bond Life Sciences Center, University of Missouri, Columbia, MO 65201, USA. AD - Department of Molecular Microbiology & Immunology, University of Missouri School of Medicine, Columbia, MO 65212, USA. FAU - Ji, Juan AU - Ji J AD - CS Bond Life Sciences Center, University of Missouri, Columbia, MO 65201, USA. FAU - Michailidis, Eleftherios AU - Michailidis E AUID- ORCID: 0000-0002-9907-4346 AD - Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA. FAU - Saeed, Mohsan AU - Saeed M AD - Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA. FAU - Ndongwe, Tanyaradzwa P AU - Ndongwe TP AD - CS Bond Life Sciences Center, University of Missouri, Columbia, MO 65201, USA. AD - Department of Molecular Microbiology & Immunology, University of Missouri School of Medicine, Columbia, MO 65212, USA. FAU - Bassit, Leda C AU - Bassit LC AD - Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, USA. FAU - Schinazi, Raymond F AU - Schinazi RF AD - Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, USA. FAU - Ralston, Robert AU - Ralston R AD - CS Bond Life Sciences Center, University of Missouri, Columbia, MO 65201, USA. AD - Department of Molecular Microbiology & Immunology, University of Missouri School of Medicine, Columbia, MO 65212, USA. FAU - Rice, Charles M AU - Rice CM AD - Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA. FAU - Sarafianos, Stefan G AU - Sarafianos SG AD - CS Bond Life Sciences Center, University of Missouri, Columbia, MO 65201, USA. AD - Department of Molecular Microbiology & Immunology, University of Missouri School of Medicine, Columbia, MO 65212, USA. AD - Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, USA. AD - Department of Biochemistry, University of Missouri, Columbia, MO 65201, USA. LA - eng GR - AI120860/AI/NIAID NIH HHS/United States GR - P30 AI050409/AI/NIAID NIH HHS/United States GR - 2P30-AI-050409/AI/NIAID NIH HHS/United States GR - AI099284/AI/NIAID NIH HHS/United States GR - P50 GM103368/GM/NIGMS NIH HHS/United States GR - AI121315/AI/NIAID NIH HHS/United States GR - R01 AI120860/AI/NIAID NIH HHS/United States GR - R01 AI099284/AI/NIAID NIH HHS/United States GR - GM103368/GM/NIGMS NIH HHS/United States GR - R01 AI121315/AI/NIAID NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural DEP - 20191108 PL - Switzerland TA - Viruses JT - Viruses JID - 101509722 RN - 0 (Antiviral Agents) RN - 0 (RNA, Viral) SB - IM MH - Antiviral Agents/*pharmacology MH - Cell Line MH - *Hepacivirus/drug effects/genetics MH - Hepatitis C/drug therapy/virology MH - Humans MH - In Situ Hybridization, Fluorescence/methods MH - *RNA, Viral/drug effects/metabolism MH - Virus Replication/drug effects MH - Zika Virus/drug effects/genetics MH - Zika Virus Infection/drug therapy/virology PMC - PMC6893808 OTO - NOTNLM OT - RNA FISH OT - Zika virus OT - direct acting antivirals OT - hepatitis C virus OT - negative-sense OT - positive-sense COIS- The authors declare no conflict of interest. EDAT- 2019/11/14 06:00 MHDA- 2020/09/30 06:00 PMCR- 2019/11/01 CRDT- 2019/11/14 06:00 PHST- 2019/10/11 00:00 [received] PHST- 2019/11/03 00:00 [revised] PHST- 2019/11/07 00:00 [accepted] PHST- 2019/11/14 06:00 [entrez] PHST- 2019/11/14 06:00 [pubmed] PHST- 2020/09/30 06:00 [medline] PHST- 2019/11/01 00:00 [pmc-release] AID - v11111039 [pii] AID - viruses-11-01039 [pii] AID - 10.3390/v11111039 [doi] PST - epublish SO - Viruses. 2019 Nov 8;11(11):1039. doi: 10.3390/v11111039.