Inhibition of Coxsackievirus B3 cardiotropic strain Woodruff replication by silencing essential viral genes
Post-transcriptional gene silencing is an effective tool for viral replication control at the epigenetic level. In this study, we aimed to produce dsRNAs and siRNA pools targeting the genetic region for the viral protease 3C and the genetic region covering the sequences for VP1 and VP3 capsid protei...
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Online Access: | http://dx.doi.org/10.1080/13102818.2019.1680318 |
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doaj-1b7100f97dc848d1a12d57398865cc3a2020-11-24T21:25:07ZengTaylor & Francis GroupBiotechnology & Biotechnological Equipment1310-28181314-35302019-01-013311582158910.1080/13102818.2019.16803181680318Inhibition of Coxsackievirus B3 cardiotropic strain Woodruff replication by silencing essential viral genesNikolay Petrov0Mariya Stoyanova1Angel Galabov2New Bulgarian UniversityLaboratory of Molecular Biology, Institute of Soil Science, Agrotechnologies and Plant Protection ‘Nikola Pushkarov’, Agricultural AcademyThe Stephan Angeloff Institute of Microbiology, Bulgarian Academy of SciencesPost-transcriptional gene silencing is an effective tool for viral replication control at the epigenetic level. In this study, we aimed to produce dsRNAs and siRNA pools targeting the genetic region for the viral protease 3C and the genetic region covering the sequences for VP1 and VP3 capsid proteins of the Coxsackievirus B3 cardiotropic Woodruff strain by the use of the polymerase complex of bacteriophage Phi6 in HEp-2 monolayer cells. The dsRNAs obtained in this study, specific for the 3C and VP1-VP3 genetic regions of the virus, were characterized by very low cytotoxicity (up to 200 nmol/L and 100 nmol/L, respectively) and high efficiency at silencing the target genes and blocking the enteroviral replication in vitro (93% and 60% reduction of the virus infected cells, respectively, at concentrations of 40 nmol/L). The generated pools of overlapping siRNAs targeting the 3C or VP1-VP3 genetic regions showed insignificant cytotoxicity at concentrations up to 200 nmol/L (< 3%) and higher efficacy than the corresponding dsRNAs. We managed to achieve 95% inhibition of the CVB3 Woodruff strain replication in vitro by using siRNAs specific for the 3C coding region of the virus at a concentration of 40 nmol/L, with no cytotoxicity.http://dx.doi.org/10.1080/13102818.2019.1680318ptgsdsrnassirnascoxsackievirus b3cvbenterovirushep-2 cellsreplication inhibition |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Nikolay Petrov Mariya Stoyanova Angel Galabov |
spellingShingle |
Nikolay Petrov Mariya Stoyanova Angel Galabov Inhibition of Coxsackievirus B3 cardiotropic strain Woodruff replication by silencing essential viral genes Biotechnology & Biotechnological Equipment ptgs dsrnas sirnas coxsackievirus b3 cvb enterovirus hep-2 cells replication inhibition |
author_facet |
Nikolay Petrov Mariya Stoyanova Angel Galabov |
author_sort |
Nikolay Petrov |
title |
Inhibition of Coxsackievirus B3 cardiotropic strain Woodruff replication by silencing essential viral genes |
title_short |
Inhibition of Coxsackievirus B3 cardiotropic strain Woodruff replication by silencing essential viral genes |
title_full |
Inhibition of Coxsackievirus B3 cardiotropic strain Woodruff replication by silencing essential viral genes |
title_fullStr |
Inhibition of Coxsackievirus B3 cardiotropic strain Woodruff replication by silencing essential viral genes |
title_full_unstemmed |
Inhibition of Coxsackievirus B3 cardiotropic strain Woodruff replication by silencing essential viral genes |
title_sort |
inhibition of coxsackievirus b3 cardiotropic strain woodruff replication by silencing essential viral genes |
publisher |
Taylor & Francis Group |
series |
Biotechnology & Biotechnological Equipment |
issn |
1310-2818 1314-3530 |
publishDate |
2019-01-01 |
description |
Post-transcriptional gene silencing is an effective tool for viral replication control at the epigenetic level. In this study, we aimed to produce dsRNAs and siRNA pools targeting the genetic region for the viral protease 3C and the genetic region covering the sequences for VP1 and VP3 capsid proteins of the Coxsackievirus B3 cardiotropic Woodruff strain by the use of the polymerase complex of bacteriophage Phi6 in HEp-2 monolayer cells. The dsRNAs obtained in this study, specific for the 3C and VP1-VP3 genetic regions of the virus, were characterized by very low cytotoxicity (up to 200 nmol/L and 100 nmol/L, respectively) and high efficiency at silencing the target genes and blocking the enteroviral replication in vitro (93% and 60% reduction of the virus infected cells, respectively, at concentrations of 40 nmol/L). The generated pools of overlapping siRNAs targeting the 3C or VP1-VP3 genetic regions showed insignificant cytotoxicity at concentrations up to 200 nmol/L (< 3%) and higher efficacy than the corresponding dsRNAs. We managed to achieve 95% inhibition of the CVB3 Woodruff strain replication in vitro by using siRNAs specific for the 3C coding region of the virus at a concentration of 40 nmol/L, with no cytotoxicity. |
topic |
ptgs dsrnas sirnas coxsackievirus b3 cvb enterovirus hep-2 cells replication inhibition |
url |
http://dx.doi.org/10.1080/13102818.2019.1680318 |
work_keys_str_mv |
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