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Visualization of Positive and Negative Sense Viral RNA for Probing the Mechanism of Direct-Acting Antivirals against Hepatitis C Virus

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 ho...

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Autores principales: Liu, Dandan, Tedbury, Philip R., Lan, Shuiyun, Huber, Andrew D., Puray-Chavez, Maritza N., Ji, Juan, Michailidis, Eleftherios, Saeed, Mohsan, Ndongwe, Tanyaradzwa P., Bassit, Leda C., Schinazi, Raymond F., Ralston, Robert, Rice, Charles M., Sarafianos, Stefan G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6893808/
https://www.ncbi.nlm.nih.gov/pubmed/31717338
http://dx.doi.org/10.3390/v11111039
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author Liu, Dandan
Tedbury, Philip R.
Lan, Shuiyun
Huber, Andrew D.
Puray-Chavez, Maritza N.
Ji, Juan
Michailidis, Eleftherios
Saeed, Mohsan
Ndongwe, Tanyaradzwa P.
Bassit, Leda C.
Schinazi, Raymond F.
Ralston, Robert
Rice, Charles M.
Sarafianos, Stefan G.
author_facet Liu, Dandan
Tedbury, Philip R.
Lan, Shuiyun
Huber, Andrew D.
Puray-Chavez, Maritza N.
Ji, Juan
Michailidis, Eleftherios
Saeed, Mohsan
Ndongwe, Tanyaradzwa P.
Bassit, Leda C.
Schinazi, Raymond F.
Ralston, Robert
Rice, Charles M.
Sarafianos, Stefan G.
author_sort Liu, Dandan
collection PubMed
description 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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spelling pubmed-68938082019-12-23 Visualization of Positive and Negative Sense Viral RNA for Probing the Mechanism of Direct-Acting Antivirals against Hepatitis C Virus Liu, Dandan Tedbury, Philip R. Lan, Shuiyun Huber, Andrew D. Puray-Chavez, Maritza N. Ji, Juan Michailidis, Eleftherios Saeed, Mohsan Ndongwe, Tanyaradzwa P. Bassit, Leda C. Schinazi, Raymond F. Ralston, Robert Rice, Charles M. Sarafianos, Stefan G. Viruses Article 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. MDPI 2019-11-08 /pmc/articles/PMC6893808/ /pubmed/31717338 http://dx.doi.org/10.3390/v11111039 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Liu, Dandan
Tedbury, Philip R.
Lan, Shuiyun
Huber, Andrew D.
Puray-Chavez, Maritza N.
Ji, Juan
Michailidis, Eleftherios
Saeed, Mohsan
Ndongwe, Tanyaradzwa P.
Bassit, Leda C.
Schinazi, Raymond F.
Ralston, Robert
Rice, Charles M.
Sarafianos, Stefan G.
Visualization of Positive and Negative Sense Viral RNA for Probing the Mechanism of Direct-Acting Antivirals against Hepatitis C Virus
title Visualization of Positive and Negative Sense Viral RNA for Probing the Mechanism of Direct-Acting Antivirals against Hepatitis C Virus
title_full Visualization of Positive and Negative Sense Viral RNA for Probing the Mechanism of Direct-Acting Antivirals against Hepatitis C Virus
title_fullStr Visualization of Positive and Negative Sense Viral RNA for Probing the Mechanism of Direct-Acting Antivirals against Hepatitis C Virus
title_full_unstemmed Visualization of Positive and Negative Sense Viral RNA for Probing the Mechanism of Direct-Acting Antivirals against Hepatitis C Virus
title_short Visualization of Positive and Negative Sense Viral RNA for Probing the Mechanism of Direct-Acting Antivirals against Hepatitis C Virus
title_sort visualization of positive and negative sense viral rna for probing the mechanism of direct-acting antivirals against hepatitis c virus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6893808/
https://www.ncbi.nlm.nih.gov/pubmed/31717338
http://dx.doi.org/10.3390/v11111039
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