tetano
Editor, Senior Moderator
Antiviral Res. 2013 Sep 18. pii: S0166-3542(13)00250-7. doi: 10.1016/j.antiviral.2013.09.004. [Epub ahead of print]
Single-stranded DNA aptamer that specifically binds to the influenza virus NS1 protein suppresses interferon antagonism.
Woo HM, Kim KS, Lee JM, Shim HS, Cho SJ, Lee WK, Ko HW, Keum YS, Kim SY, Pathinayake P, Kim CJ, Jeong YJ.
Source
Department of Bio and Nanochemistry, Kookmin University, Seoul 136-702, Republic of Korea.
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.
Copyright ? 2013. Published by Elsevier B.V.
KEYWORDS:
Aptamer, G-quadruplex, Influenza Virus, Interferon, NS1
PMID:
24055449
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24055449
Single-stranded DNA aptamer that specifically binds to the influenza virus NS1 protein suppresses interferon antagonism.
Woo HM, Kim KS, Lee JM, Shim HS, Cho SJ, Lee WK, Ko HW, Keum YS, Kim SY, Pathinayake P, Kim CJ, Jeong YJ.
Source
Department of Bio and Nanochemistry, Kookmin University, Seoul 136-702, Republic of Korea.
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.
Copyright ? 2013. Published by Elsevier B.V.
KEYWORDS:
Aptamer, G-quadruplex, Influenza Virus, Interferon, NS1
PMID:
24055449
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24055449