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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
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.
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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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