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Characterization of DNA aptamers generated against the soft-shelled turtle iridovirus with antiviral effects

机译:针对甲鱼虹膜病毒产生的具有抗病毒作用的DNA适体的表征

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Background Soft-shelled turtle iridovirus (STIV) causes severe systemic disease in farmed soft-shelled turtles ( Trionyx sinensis ). More efficient methods of controlling and detecting STIV infections are urgently needed. Methods In this study, we generated eight single-stranded DNA (ssDNA) aptamers against STIV using systematic evolution of ligands by exponential enrichment (SELEX). Results The aptamers formed representative stem-loop secondary structures. Electrophoretic mobility shift assays?and fluorescent localization showed that the selected aptamers had high binding affinity for STIV. Aptamer?QA-36 had the highest calculated binding affinity ( K d ) of 53.8 nM. Flow cytometry and fluorescence microscopy of?cell-aptamer interactions demonstrated that QA-12 was able to recognize both STIV-infected cells and tissues with a?high level of specificity. Moreover, the selected aptamers inhibited STIV infection in vitro and in vivo , with aptamer QA-36 demonstrating the greatest protective effect against STIV and inhibiting STIV infection in a dose-dependent manner. Discussion We generated DNA aptamers that bound STIV with a high level of specificity, providing an alternative means for investigating STIV pathogenesis, drug development, and medical therapies for STIV infection. Conclusions These DNA aptamers may thus be suitable antiviral candidates for the control of STIV infections.
机译:背景甲鱼虹膜病毒(STIV)在养殖的甲鱼(Trionyx sinensis)中引起严重的全身性疾病。迫切需要更有效的控制和检测STIV感染的方法。方法在本研究中,我们利用配体通过指数富集(SELEX)的系统进化,生成了8种抗STIV的单链DNA(ssDNA)适体。结果适体形成了代表性的茎环二级结构。电泳迁移率迁移分析和荧光定位表明,所选择的适体对STIV具有高结合亲和力。 Aptamer?QA-36的最高计算结合亲和力(K d )为53.8 nM。细胞-适体相互作用的流式细胞术和荧光显微镜检查表明,QA-12能够以很高的特异性识别被STIV感染的细胞和组织。此外,选择的适体在体外和体内均抑制STIV感染,而适体QA-36显示出对STIV的最大保护作用并以剂量依赖性方式抑制STIV感染。讨论我们产生了以高度特异性结合STIV的DNA适体,为研究STIV的发病机理,药物开发和STIV感染的药物治疗提供了另一种方法。结论这些DNA适体因此可能是控制STIV感染的合适抗病毒候选物。

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