Single-stranded DNA aptamer that specifically binds to the influenza virus NS1 protein suppresses interferon antagonism

Hye Min Woo, Ki Sun Kim, Jin Moo Lee, Hee Sup Shim, Seong Je Cho, Won Kyu Lee, Hyuk Wan Ko, Young Sam Keum, Soo Youl Kim, Prabuddha Pathinayake, Chul Joong Kim, Yong Joo Jeong

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Non-structural protein 1 (NS1) of the influenza A virus (IAV) inhibits the host's innate immune response by suppressing the induction of interferons (IFNs). Therefore, blocking NS1 activity can be a potential strategy in the development of antiviral agents against IAV infection. In the present study, we selected a single-stranded DNA aptamer specific to the IAV NS1 protein after 15 cycles of systematic evolution of ligands by exponential enrichment (SELEX) procedure and examined the ability of the selected aptamer to inhibit the function of NS1. The selected aptamer binds to NS1 with a Kd of 18.91 ± 3.95 nM and RNA binding domain of NS1 is determined to be critical for the aptamer binding. The aptamer has a G-rich sequence in the random sequence region and forms a G-quadruplex structure. The localization of the aptamer bound to NS1 in cells was determined by confocal images, and flow cytometry analysis further demonstrated that the selected aptamer binds specifically to NS1. In addition, luciferase reporter gene assay, quantitative RT-PCR, and enzyme-linked immunosorbent assay (ELISA) experiments demonstrated that the selected aptamer had the ability to induce IFN-β by suppressing the function of NS1. Importantly, we also found that the selected aptamer was able to inhibit the viral replication without affecting cell viability. These results indicate that the selected ssDNA aptamer has strong potential to be further developed as a therapeutic agent against IAV.

Original languageEnglish
Pages (from-to)337-345
Number of pages9
JournalAntiviral Research
Volume100
Issue number2
DOIs
Publication statusPublished - 2013 Oct 16

Fingerprint

Nucleotide Aptamers
Viral Structural Proteins
Single-Stranded DNA
Orthomyxoviridae
Interferons
Influenza A virus
Proteins
SELEX Aptamer Technique
Image Cytometry
G-Quadruplexes
Virus Diseases
Luciferases
Reporter Genes
Innate Immunity
Antiviral Agents
Cell Survival
Flow Cytometry
Enzyme-Linked Immunosorbent Assay

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Virology

Cite this

Woo, Hye Min ; Kim, Ki Sun ; Lee, Jin Moo ; Shim, Hee Sup ; Cho, Seong Je ; Lee, Won Kyu ; Ko, Hyuk Wan ; Keum, Young Sam ; Kim, Soo Youl ; Pathinayake, Prabuddha ; Kim, Chul Joong ; Jeong, Yong Joo. / Single-stranded DNA aptamer that specifically binds to the influenza virus NS1 protein suppresses interferon antagonism. In: Antiviral Research. 2013 ; Vol. 100, No. 2. pp. 337-345.
@article{343b77eba5db45cda6ccf2d33c0f0947,
title = "Single-stranded DNA aptamer that specifically binds to the influenza virus NS1 protein suppresses interferon antagonism",
abstract = "Non-structural protein 1 (NS1) of the influenza A virus (IAV) inhibits the host's innate immune response by suppressing the induction of interferons (IFNs). Therefore, blocking NS1 activity can be a potential strategy in the development of antiviral agents against IAV infection. In the present study, we selected a single-stranded DNA aptamer specific to the IAV NS1 protein after 15 cycles of systematic evolution of ligands by exponential enrichment (SELEX) procedure and examined the ability of the selected aptamer to inhibit the function of NS1. The selected aptamer binds to NS1 with a Kd of 18.91 ± 3.95 nM and RNA binding domain of NS1 is determined to be critical for the aptamer binding. The aptamer has a G-rich sequence in the random sequence region and forms a G-quadruplex structure. The localization of the aptamer bound to NS1 in cells was determined by confocal images, and flow cytometry analysis further demonstrated that the selected aptamer binds specifically to NS1. In addition, luciferase reporter gene assay, quantitative RT-PCR, and enzyme-linked immunosorbent assay (ELISA) experiments demonstrated that the selected aptamer had the ability to induce IFN-β by suppressing the function of NS1. Importantly, we also found that the selected aptamer was able to inhibit the viral replication without affecting cell viability. These results indicate that the selected ssDNA aptamer has strong potential to be further developed as a therapeutic agent against IAV.",
author = "Woo, {Hye Min} and Kim, {Ki Sun} and Lee, {Jin Moo} and Shim, {Hee Sup} and Cho, {Seong Je} and Lee, {Won Kyu} and Ko, {Hyuk Wan} and Keum, {Young Sam} and Kim, {Soo Youl} and Prabuddha Pathinayake and Kim, {Chul Joong} and Jeong, {Yong Joo}",
year = "2013",
month = "10",
day = "16",
doi = "10.1016/j.antiviral.2013.09.004",
language = "English",
volume = "100",
pages = "337--345",
journal = "Antiviral Research",
issn = "0166-3542",
publisher = "Elsevier",
number = "2",

}

Woo, HM, Kim, KS, Lee, JM, Shim, HS, Cho, SJ, Lee, WK, Ko, HW, Keum, YS, Kim, SY, Pathinayake, P, Kim, CJ & Jeong, YJ 2013, 'Single-stranded DNA aptamer that specifically binds to the influenza virus NS1 protein suppresses interferon antagonism', Antiviral Research, vol. 100, no. 2, pp. 337-345. https://doi.org/10.1016/j.antiviral.2013.09.004

Single-stranded DNA aptamer that specifically binds to the influenza virus NS1 protein suppresses interferon antagonism. / Woo, Hye Min; Kim, Ki Sun; Lee, Jin Moo; Shim, Hee Sup; Cho, Seong Je; Lee, Won Kyu; Ko, Hyuk Wan; Keum, Young Sam; Kim, Soo Youl; Pathinayake, Prabuddha; Kim, Chul Joong; Jeong, Yong Joo.

In: Antiviral Research, Vol. 100, No. 2, 16.10.2013, p. 337-345.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Single-stranded DNA aptamer that specifically binds to the influenza virus NS1 protein suppresses interferon antagonism

AU - Woo, Hye Min

AU - Kim, Ki Sun

AU - Lee, Jin Moo

AU - Shim, Hee Sup

AU - Cho, Seong Je

AU - Lee, Won Kyu

AU - Ko, Hyuk Wan

AU - Keum, Young Sam

AU - Kim, Soo Youl

AU - Pathinayake, Prabuddha

AU - Kim, Chul Joong

AU - Jeong, Yong Joo

PY - 2013/10/16

Y1 - 2013/10/16

N2 - Non-structural protein 1 (NS1) of the influenza A virus (IAV) inhibits the host's innate immune response by suppressing the induction of interferons (IFNs). Therefore, blocking NS1 activity can be a potential strategy in the development of antiviral agents against IAV infection. In the present study, we selected a single-stranded DNA aptamer specific to the IAV NS1 protein after 15 cycles of systematic evolution of ligands by exponential enrichment (SELEX) procedure and examined the ability of the selected aptamer to inhibit the function of NS1. The selected aptamer binds to NS1 with a Kd of 18.91 ± 3.95 nM and RNA binding domain of NS1 is determined to be critical for the aptamer binding. The aptamer has a G-rich sequence in the random sequence region and forms a G-quadruplex structure. The localization of the aptamer bound to NS1 in cells was determined by confocal images, and flow cytometry analysis further demonstrated that the selected aptamer binds specifically to NS1. In addition, luciferase reporter gene assay, quantitative RT-PCR, and enzyme-linked immunosorbent assay (ELISA) experiments demonstrated that the selected aptamer had the ability to induce IFN-β by suppressing the function of NS1. Importantly, we also found that the selected aptamer was able to inhibit the viral replication without affecting cell viability. These results indicate that the selected ssDNA aptamer has strong potential to be further developed as a therapeutic agent against IAV.

AB - Non-structural protein 1 (NS1) of the influenza A virus (IAV) inhibits the host's innate immune response by suppressing the induction of interferons (IFNs). Therefore, blocking NS1 activity can be a potential strategy in the development of antiviral agents against IAV infection. In the present study, we selected a single-stranded DNA aptamer specific to the IAV NS1 protein after 15 cycles of systematic evolution of ligands by exponential enrichment (SELEX) procedure and examined the ability of the selected aptamer to inhibit the function of NS1. The selected aptamer binds to NS1 with a Kd of 18.91 ± 3.95 nM and RNA binding domain of NS1 is determined to be critical for the aptamer binding. The aptamer has a G-rich sequence in the random sequence region and forms a G-quadruplex structure. The localization of the aptamer bound to NS1 in cells was determined by confocal images, and flow cytometry analysis further demonstrated that the selected aptamer binds specifically to NS1. In addition, luciferase reporter gene assay, quantitative RT-PCR, and enzyme-linked immunosorbent assay (ELISA) experiments demonstrated that the selected aptamer had the ability to induce IFN-β by suppressing the function of NS1. Importantly, we also found that the selected aptamer was able to inhibit the viral replication without affecting cell viability. These results indicate that the selected ssDNA aptamer has strong potential to be further developed as a therapeutic agent against IAV.

UR - http://www.scopus.com/inward/record.url?scp=84885340369&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84885340369&partnerID=8YFLogxK

U2 - 10.1016/j.antiviral.2013.09.004

DO - 10.1016/j.antiviral.2013.09.004

M3 - Article

VL - 100

SP - 337

EP - 345

JO - Antiviral Research

JF - Antiviral Research

SN - 0166-3542

IS - 2

ER -