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Int. J. Mol. Sci. 2016, 17(2), 154;

Identification and Characterization of MicroRNAs in Snakehead Fish Cell Line upon Snakehead Fish Vesiculovirus Infection

Department of Aquatic Animal Medicine, College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China
Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan 430070, China
Department of Zoology, Hazara University, Mansehra, Khyber Pakhtoonkhwa 21300, Pakistan
Central Laboratory for Environmental Quality Monitoring (CLEQM), National Water Research Center (NWRC), Cairo 13621, Egypt
Institute of Marine and Environmental Technology, University of Maryland, Baltimore, MD 21202, USA
These authors contributed equally to the work.
Authors to whom correspondence should be addressed.
Academic Editor: Jun Li
Received: 5 January 2016 / Revised: 19 January 2016 / Accepted: 21 January 2016 / Published: 26 January 2016
(This article belongs to the Special Issue Fish Molecular Biology)
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MicroRNAs (miRNAs) play important roles in mediating multiple biological processes in eukaryotes and are being increasingly studied to evaluate their roles associated with cellular changes following viral infection. Snakehead fish Vesiculovirus (SHVV) has caused mass mortality in snakehead fish during the past few years. To identify specific miRNAs involved in SHVV infection, we performed microRNA deep sequencing on a snakehead fish cell line (SSN-1) with or without SHVV infection. A total of 205 known miRNAs were identified when they were aligned with the known zebrafish miRNAs, and nine novel miRNAs were identified using MiRDeep2 software. Eighteen and 143 of the 205 known miRNAs were differentially expressed at three and 24 h post-infection (poi), respectively. From the differentially-expressed miRNAs, five were randomly selected to validate their expression profiles using quantitative reverse transcription polymerase chain reaction (qRT-PCR), and their expression profiles were consistent with the microRNA sequencing results. In addition, the target gene prediction of the SHVV genome was performed for the differentially-expressed host miRNAs, and a total of 10 and 58 differentially-expressed miRNAs were predicted to bind to the SHVV genome at three and 24 h poi, respectively. The effects of three selected miRNAs (miR-130-5p, miR-214 and miR-216b) on SHVV multiplication were evaluated using their mimics and inhibitors via qRT-PCR and Western blotting. The results showed that all three miRNAs were able to inhibit the multiplication of SHVV; whereas the mechanisms underlying the SHVV multiplication inhibited by the specific miRNAs need to be further characterized in the future. View Full-Text
Keywords: snakehead fish Vesiculovirus; SSN-1 cell; miRNA; deep sequencing snakehead fish Vesiculovirus; SSN-1 cell; miRNA; deep sequencing

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Liu, X.; Tu, J.; Yuan, J.; Liu, X.; Zhao, L.; Dawar, F.U.; Khattak, M.N.K.; Hegazy, A.M.; Chen, N.; Vakharia, V.N.; Lin, L. Identification and Characterization of MicroRNAs in Snakehead Fish Cell Line upon Snakehead Fish Vesiculovirus Infection. Int. J. Mol. Sci. 2016, 17, 154.

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