RNA Aptamers Inhibit the Growth of the Fish Pathogen Viral Hemorrhagic Septicemia Virus (VHSV)

Porntep Punnarak, Mudjekeewis D. Santos, Seong Don Hwang, Hidehiro Kondo, Ikuo Hirono, Yo Kikuchi, Takashi Aoki

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Viral hemorrhagic septicemia virus (VHSV) is a serious disease impacting wild and cultured fish worldwide. Hence, an effective therapeutic method against VHSV infection needs to be developed. Aptamer technology is a new and promising method for diagnostics and therapeutics. It revolves around the use of an aptamer molecule, an artificial ligand (nucleic acid or protein), which has the capacity to recognize target molecules with high affinity and specificity. Here, we aimed at selecting RNA aptamers that can specifically bind to and inhibit the growth of a strain of fish VHSV both in vitro and in vivo. Three VHSV-specific RNA aptamers (F1, F2, and C6) were selected from a pool of artificially and randomly produced oligonucleotides using systematic evolution of ligands by exponential enrichment. The three RNA aptamers showed obvious binding to VHSV in an electrophoretic mobility shift assay but not to other tested viruses. The RNA aptamers were tested for their ability to inhibit VHSV in vitro using hirame natural embryo (HINAE) cells. Cytopathic effect and plaque assays showed that all aptamers inhibited the growth of VHSV in HINAE cells. In vivo tests using RNA aptamers produced by Rhodovulum sulfidophilum showed that extracellular RNA aptamers inhibited VHSV infection in Japanese flounder. These results suggest that the RNA aptamers are a useful tool for protection against VHSV infection in Japanese flounder.

Original languageEnglish
Pages (from-to)752-761
Number of pages10
JournalMarine Biotechnology
Volume14
Issue number6
DOIs
Publication statusPublished - 2012
Externally publishedYes

Fingerprint

Novirhabdovirus
Nucleotide Aptamers
Viral hemorrhagic septicemia virus
RNA
virus
pathogen
pathogens
fish
Growth
Virus Diseases
Flounder
Paralichthys olivaceus
Rhodovulum
Rhodovulum sulfidophilum
embryo (animal)
ligand
Fishes
SELEX Aptamer Technique
Embryonic Structures
embryo

Keywords

  • Japanese flounder
  • Rhodovulum sulfidophilum
  • RNA aptamer
  • VHSV

ASJC Scopus subject areas

  • Aquatic Science
  • Biotechnology

Cite this

Punnarak, P., Santos, M. D., Hwang, S. D., Kondo, H., Hirono, I., Kikuchi, Y., & Aoki, T. (2012). RNA Aptamers Inhibit the Growth of the Fish Pathogen Viral Hemorrhagic Septicemia Virus (VHSV). Marine Biotechnology, 14(6), 752-761. https://doi.org/10.1007/s10126-012-9448-1

RNA Aptamers Inhibit the Growth of the Fish Pathogen Viral Hemorrhagic Septicemia Virus (VHSV). / Punnarak, Porntep; Santos, Mudjekeewis D.; Hwang, Seong Don; Kondo, Hidehiro; Hirono, Ikuo; Kikuchi, Yo; Aoki, Takashi.

In: Marine Biotechnology, Vol. 14, No. 6, 2012, p. 752-761.

Research output: Contribution to journalArticle

Punnarak, P, Santos, MD, Hwang, SD, Kondo, H, Hirono, I, Kikuchi, Y & Aoki, T 2012, 'RNA Aptamers Inhibit the Growth of the Fish Pathogen Viral Hemorrhagic Septicemia Virus (VHSV)', Marine Biotechnology, vol. 14, no. 6, pp. 752-761. https://doi.org/10.1007/s10126-012-9448-1
Punnarak, Porntep ; Santos, Mudjekeewis D. ; Hwang, Seong Don ; Kondo, Hidehiro ; Hirono, Ikuo ; Kikuchi, Yo ; Aoki, Takashi. / RNA Aptamers Inhibit the Growth of the Fish Pathogen Viral Hemorrhagic Septicemia Virus (VHSV). In: Marine Biotechnology. 2012 ; Vol. 14, No. 6. pp. 752-761.
@article{4788a17a1abe4e269fa20cf79b66c57a,
title = "RNA Aptamers Inhibit the Growth of the Fish Pathogen Viral Hemorrhagic Septicemia Virus (VHSV)",
abstract = "Viral hemorrhagic septicemia virus (VHSV) is a serious disease impacting wild and cultured fish worldwide. Hence, an effective therapeutic method against VHSV infection needs to be developed. Aptamer technology is a new and promising method for diagnostics and therapeutics. It revolves around the use of an aptamer molecule, an artificial ligand (nucleic acid or protein), which has the capacity to recognize target molecules with high affinity and specificity. Here, we aimed at selecting RNA aptamers that can specifically bind to and inhibit the growth of a strain of fish VHSV both in vitro and in vivo. Three VHSV-specific RNA aptamers (F1, F2, and C6) were selected from a pool of artificially and randomly produced oligonucleotides using systematic evolution of ligands by exponential enrichment. The three RNA aptamers showed obvious binding to VHSV in an electrophoretic mobility shift assay but not to other tested viruses. The RNA aptamers were tested for their ability to inhibit VHSV in vitro using hirame natural embryo (HINAE) cells. Cytopathic effect and plaque assays showed that all aptamers inhibited the growth of VHSV in HINAE cells. In vivo tests using RNA aptamers produced by Rhodovulum sulfidophilum showed that extracellular RNA aptamers inhibited VHSV infection in Japanese flounder. These results suggest that the RNA aptamers are a useful tool for protection against VHSV infection in Japanese flounder.",
keywords = "Japanese flounder, Rhodovulum sulfidophilum, RNA aptamer, VHSV",
author = "Porntep Punnarak and Santos, {Mudjekeewis D.} and Hwang, {Seong Don} and Hidehiro Kondo and Ikuo Hirono and Yo Kikuchi and Takashi Aoki",
year = "2012",
doi = "10.1007/s10126-012-9448-1",
language = "English",
volume = "14",
pages = "752--761",
journal = "Marine Biotechnology",
issn = "1436-2228",
publisher = "Springer New York",
number = "6",

}

TY - JOUR

T1 - RNA Aptamers Inhibit the Growth of the Fish Pathogen Viral Hemorrhagic Septicemia Virus (VHSV)

AU - Punnarak, Porntep

AU - Santos, Mudjekeewis D.

AU - Hwang, Seong Don

AU - Kondo, Hidehiro

AU - Hirono, Ikuo

AU - Kikuchi, Yo

AU - Aoki, Takashi

PY - 2012

Y1 - 2012

N2 - Viral hemorrhagic septicemia virus (VHSV) is a serious disease impacting wild and cultured fish worldwide. Hence, an effective therapeutic method against VHSV infection needs to be developed. Aptamer technology is a new and promising method for diagnostics and therapeutics. It revolves around the use of an aptamer molecule, an artificial ligand (nucleic acid or protein), which has the capacity to recognize target molecules with high affinity and specificity. Here, we aimed at selecting RNA aptamers that can specifically bind to and inhibit the growth of a strain of fish VHSV both in vitro and in vivo. Three VHSV-specific RNA aptamers (F1, F2, and C6) were selected from a pool of artificially and randomly produced oligonucleotides using systematic evolution of ligands by exponential enrichment. The three RNA aptamers showed obvious binding to VHSV in an electrophoretic mobility shift assay but not to other tested viruses. The RNA aptamers were tested for their ability to inhibit VHSV in vitro using hirame natural embryo (HINAE) cells. Cytopathic effect and plaque assays showed that all aptamers inhibited the growth of VHSV in HINAE cells. In vivo tests using RNA aptamers produced by Rhodovulum sulfidophilum showed that extracellular RNA aptamers inhibited VHSV infection in Japanese flounder. These results suggest that the RNA aptamers are a useful tool for protection against VHSV infection in Japanese flounder.

AB - Viral hemorrhagic septicemia virus (VHSV) is a serious disease impacting wild and cultured fish worldwide. Hence, an effective therapeutic method against VHSV infection needs to be developed. Aptamer technology is a new and promising method for diagnostics and therapeutics. It revolves around the use of an aptamer molecule, an artificial ligand (nucleic acid or protein), which has the capacity to recognize target molecules with high affinity and specificity. Here, we aimed at selecting RNA aptamers that can specifically bind to and inhibit the growth of a strain of fish VHSV both in vitro and in vivo. Three VHSV-specific RNA aptamers (F1, F2, and C6) were selected from a pool of artificially and randomly produced oligonucleotides using systematic evolution of ligands by exponential enrichment. The three RNA aptamers showed obvious binding to VHSV in an electrophoretic mobility shift assay but not to other tested viruses. The RNA aptamers were tested for their ability to inhibit VHSV in vitro using hirame natural embryo (HINAE) cells. Cytopathic effect and plaque assays showed that all aptamers inhibited the growth of VHSV in HINAE cells. In vivo tests using RNA aptamers produced by Rhodovulum sulfidophilum showed that extracellular RNA aptamers inhibited VHSV infection in Japanese flounder. These results suggest that the RNA aptamers are a useful tool for protection against VHSV infection in Japanese flounder.

KW - Japanese flounder

KW - Rhodovulum sulfidophilum

KW - RNA aptamer

KW - VHSV

UR - http://www.scopus.com/inward/record.url?scp=84869147600&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84869147600&partnerID=8YFLogxK

U2 - 10.1007/s10126-012-9448-1

DO - 10.1007/s10126-012-9448-1

M3 - Article

VL - 14

SP - 752

EP - 761

JO - Marine Biotechnology

JF - Marine Biotechnology

SN - 1436-2228

IS - 6

ER -