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

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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

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All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Virology

Cite this

Woo, H. M., Kim, K. S., Lee, J. M., Shim, H. S., Cho, S. J., Lee, W. K., Ko, H. W., Keum, Y. S., Kim, S. Y., Pathinayake, P., Kim, C. J., & Jeong, Y. J. (2013). Single-stranded DNA aptamer that specifically binds to the influenza virus NS1 protein suppresses interferon antagonism. Antiviral Research, 100(2), 337-345. https://doi.org/10.1016/j.antiviral.2013.09.004