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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...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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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. |
format | Online Article Text |
id | pubmed-6893808 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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