Generation and characterization of novel DNA aptamers against coat protein of grouper nervous necrosis virus (GNNV) with antiviral activities and delivery potential in grouper cells
Zhou, Lingli; Li, Pengfei; Yang, Min; Yu, Yepin; Huang, Youhua; Wei, Jingguang; Wei, Shina; Qin, Qiwei; Wei, SN; Qin, QW (reprint author), Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Trop Marine Bioresources & Ecol, 164 West Xingang Rd, Guangzhou 510301, Guangdong, Peoples R China.
2016
发表期刊ANTIVIRAL RESEARCH
卷号129页码:104-114
摘要Nervous necrosis virus (NNV) infected larvae and juveniles of more than 50 fish species, resulting in mortality rates of greater than 95%. However, there is no efficient method to control NNV infections. Aptamers generated by selective evolution of ligands by exponential enrichment (SELEX) are short, single-stranded nucleic acid oligomers. They display a high degree of affinity and specificity for many targets, such as viruses and viral proteins. In this study, three novel DNA aptamers (A5, A10, and B11) that specifically target the coat protein (CP) of grouper nervous necrosis virus (GNNV) were selected using SELEX. Secondary structures and minimum free energy (Delta G) predictions indicated that these aptamers could form stable, secondary stem-loop structures. Electrophoretic mobility shift assays, enzyme-linked immunosorbent assays, K-d measurements, the co-localization of tetramethylrhodamine (TAMRA) labeled-aptamers with the CP and flow cytometry analysis revealed that these aptamers could specifically bind the CP with high (nanomolar) affinities. In addition, competition analysis suggested the aptamers shared some common CP binding sites with the anti-CP antibody. Moreover, all three aptamers did not show any cytotoxic effects in vitro or in vivo, and anti-viral analysis indicated the selected aptamers could inhibit NNV infection in vitro and in vivo. Compared with controls, mortality of GNNV-infected fish decreased by 40% and 80% after 10 days infection, when the GNNV was pre-incubated with the 1000 nM A10 and B11, respectively. TAMRA-labeled aptamers could bind to NNV virions and directly enter NNV-infected cells, suggesting they could be used as tracers to study the mechanism of viral infection, as well as for targeted therapy. This is the first time that aptamers targeting a viral protein of marine fish have been generated and characterized. These aptamers hold promise as diagnostic, therapeutic, and targeted drug delivery agents for controlling NNV infections. (C) 2016 Elsevier B.V. All rights reserved.
部门归属[Zhou, Lingli; Li, Pengfei; Yang, Min; Yu, Yepin; Huang, Youhua; Wei, Jingguang; Wei, Shina; Qin, Qiwei] Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Trop Marine Bioresources & Ecol, 164 West Xingang Rd, Guangzhou 510301, Guangdong, Peoples R China; [Zhou, Lingli; Li, Pengfei; Yu, Yepin] Univ Chinese Acad Sci, 19 Yuquan Rd, Beijing 100049, Peoples R China; [Qin, Qiwei] Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao, Peoples R China ; LMB
关键词Aptamer Selex Grouper Nervous Necrosis Virus Coat Protein Antiviral Activity
学科领域Pharmacology & Pharmacy ; Virology
文献类型期刊论文
条目标识符http://ir.scsio.ac.cn/handle/344004/15422
专题中科院海洋生物资源可持续利用重点实验室
通讯作者Wei, SN; Qin, QW (reprint author), Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Trop Marine Bioresources & Ecol, 164 West Xingang Rd, Guangzhou 510301, Guangdong, Peoples R China.
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Zhou, Lingli,Li, Pengfei,Yang, Min,et al. Generation and characterization of novel DNA aptamers against coat protein of grouper nervous necrosis virus (GNNV) with antiviral activities and delivery potential in grouper cells[J]. ANTIVIRAL RESEARCH,2016,129:104-114.
APA Zhou, Lingli.,Li, Pengfei.,Yang, Min.,Yu, Yepin.,Huang, Youhua.,...&Qin, QW .(2016).Generation and characterization of novel DNA aptamers against coat protein of grouper nervous necrosis virus (GNNV) with antiviral activities and delivery potential in grouper cells.ANTIVIRAL RESEARCH,129,104-114.
MLA Zhou, Lingli,et al."Generation and characterization of novel DNA aptamers against coat protein of grouper nervous necrosis virus (GNNV) with antiviral activities and delivery potential in grouper cells".ANTIVIRAL RESEARCH 129(2016):104-114.
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