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A Rapid Screen for Host-Encoded miRNAs with Inhibitory Effects against Ebola Virus Using a Transcription- and Replication-Competent Virus-Like Particle System
MicroRNAs (miRNAs) may become efficient antiviral agents against the Ebola virus (EBOV) targeting viral genomic RNAs or transcripts. We previously conducted a genome-wide search for differentially expressed miRNAs during viral replication and transcription. In this study, we established a rapid scre...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5983748/ https://www.ncbi.nlm.nih.gov/pubmed/29772717 http://dx.doi.org/10.3390/ijms19051488 |
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author | Wang, Zhongyi Li, Jiaming Fu, Yingying Zhao, Zongzheng Zhang, Chunmao Li, Nan Li, Jingjing Cheng, Hongliang Jin, Xiaojun Lu, Bing Guo, Zhendong Qian, Jun Liu, Linna |
author_facet | Wang, Zhongyi Li, Jiaming Fu, Yingying Zhao, Zongzheng Zhang, Chunmao Li, Nan Li, Jingjing Cheng, Hongliang Jin, Xiaojun Lu, Bing Guo, Zhendong Qian, Jun Liu, Linna |
author_sort | Wang, Zhongyi |
collection | PubMed |
description | MicroRNAs (miRNAs) may become efficient antiviral agents against the Ebola virus (EBOV) targeting viral genomic RNAs or transcripts. We previously conducted a genome-wide search for differentially expressed miRNAs during viral replication and transcription. In this study, we established a rapid screen for miRNAs with inhibitory effects against EBOV using a tetracistronic transcription- and replication-competent virus-like particle (trVLP) system. This system uses a minigenome comprising an EBOV leader region, luciferase reporter, VP40, GP, VP24, EBOV trailer region, and three noncoding regions from the EBOV genome and can be used to model the life cycle of EBOV under biosafety level (BSL) 2 conditions. Informatic analysis was performed to select up-regulated miRNAs targeting the coding regions of the minigenome with the highest binding energy to perform inhibitory effect screening. Among these miRNAs, miR-150-3p had the most significant inhibitory effect. Reverse transcription polymerase chain reaction (RT-PCR), Western blot, and double fluorescence reporter experiments demonstrated that miR-150-3p inhibited the reproduction of trVLPs via the regulation of GP and VP40 expression by directly targeting the coding regions of GP and VP40. This novel, rapid, and convenient screening method will efficiently facilitate the exploration of miRNAs against EBOV under BSL-2 conditions. |
format | Online Article Text |
id | pubmed-5983748 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-59837482018-06-05 A Rapid Screen for Host-Encoded miRNAs with Inhibitory Effects against Ebola Virus Using a Transcription- and Replication-Competent Virus-Like Particle System Wang, Zhongyi Li, Jiaming Fu, Yingying Zhao, Zongzheng Zhang, Chunmao Li, Nan Li, Jingjing Cheng, Hongliang Jin, Xiaojun Lu, Bing Guo, Zhendong Qian, Jun Liu, Linna Int J Mol Sci Article MicroRNAs (miRNAs) may become efficient antiviral agents against the Ebola virus (EBOV) targeting viral genomic RNAs or transcripts. We previously conducted a genome-wide search for differentially expressed miRNAs during viral replication and transcription. In this study, we established a rapid screen for miRNAs with inhibitory effects against EBOV using a tetracistronic transcription- and replication-competent virus-like particle (trVLP) system. This system uses a minigenome comprising an EBOV leader region, luciferase reporter, VP40, GP, VP24, EBOV trailer region, and three noncoding regions from the EBOV genome and can be used to model the life cycle of EBOV under biosafety level (BSL) 2 conditions. Informatic analysis was performed to select up-regulated miRNAs targeting the coding regions of the minigenome with the highest binding energy to perform inhibitory effect screening. Among these miRNAs, miR-150-3p had the most significant inhibitory effect. Reverse transcription polymerase chain reaction (RT-PCR), Western blot, and double fluorescence reporter experiments demonstrated that miR-150-3p inhibited the reproduction of trVLPs via the regulation of GP and VP40 expression by directly targeting the coding regions of GP and VP40. This novel, rapid, and convenient screening method will efficiently facilitate the exploration of miRNAs against EBOV under BSL-2 conditions. MDPI 2018-05-16 /pmc/articles/PMC5983748/ /pubmed/29772717 http://dx.doi.org/10.3390/ijms19051488 Text en © 2018 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 Wang, Zhongyi Li, Jiaming Fu, Yingying Zhao, Zongzheng Zhang, Chunmao Li, Nan Li, Jingjing Cheng, Hongliang Jin, Xiaojun Lu, Bing Guo, Zhendong Qian, Jun Liu, Linna A Rapid Screen for Host-Encoded miRNAs with Inhibitory Effects against Ebola Virus Using a Transcription- and Replication-Competent Virus-Like Particle System |
title | A Rapid Screen for Host-Encoded miRNAs with Inhibitory Effects against Ebola Virus Using a Transcription- and Replication-Competent Virus-Like Particle System |
title_full | A Rapid Screen for Host-Encoded miRNAs with Inhibitory Effects against Ebola Virus Using a Transcription- and Replication-Competent Virus-Like Particle System |
title_fullStr | A Rapid Screen for Host-Encoded miRNAs with Inhibitory Effects against Ebola Virus Using a Transcription- and Replication-Competent Virus-Like Particle System |
title_full_unstemmed | A Rapid Screen for Host-Encoded miRNAs with Inhibitory Effects against Ebola Virus Using a Transcription- and Replication-Competent Virus-Like Particle System |
title_short | A Rapid Screen for Host-Encoded miRNAs with Inhibitory Effects against Ebola Virus Using a Transcription- and Replication-Competent Virus-Like Particle System |
title_sort | rapid screen for host-encoded mirnas with inhibitory effects against ebola virus using a transcription- and replication-competent virus-like particle system |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5983748/ https://www.ncbi.nlm.nih.gov/pubmed/29772717 http://dx.doi.org/10.3390/ijms19051488 |
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