Department of Biomedical Engineering, Korea University, Hana Science Hall, 145 Anamro, Seongbukgu, Seoul 02841, Korea.
Institute of Precision Public Health, Korea University, Hana Science Hall, 145 Anamro, Seongbukgu, Seoul 02841, Korea.
Analyst. 2022 May 3;147(9):1899-1905. doi: 10.1039/d2an00209d.
Kinetically trapped hairpin DNA has great potential to dynamically build nanostructures, which can be initiated by sequence-specific nucleic acids. The branched junction, which has a multi-arm structure, is a representative nanostructure of DNA. In this study, we report a nonenzymatic and isothermal signal amplification accompanied by building a 3-arm structure based on a catalyzed hairpin DNA assembly (3-CHA). We improved the signal-to-background ratio of the 3-CHA by suppressing the leakage pathway of 3-CHA, thus eliminating unfavorable reaction sites exposed in the single-stranded region of hairpin DNAs. Background and amplified signals were analyzed with gel electrophoresis and real-time fluorescence monitoring. The limit of detection of the developed 3-CHA was estimated to be 29.3 pM for catalyst DNA at room temperature. Supported by the reduced leakage signal, the implemented 3-CHA showed great potential for detecting low concentrations of target DNA.
动力学捕获发夹 DNA 具有动态构建纳米结构的巨大潜力，可以通过序列特异性核酸来启动。具有多臂结构的分支连接是 DNA 的一种代表性纳米结构。在这项研究中，我们报告了一种非酶且等温的信号放大方法，同时构建了基于催化发夹 DNA 组装（3-CHA）的 3 臂结构。我们通过抑制 3-CHA 的漏液途径来提高 3-CHA 的信号与背景比，从而消除发夹 DNA 单链区中暴露的不利反应位点。通过凝胶电泳和实时荧光监测分析背景和放大信号。在室温下，对于催化剂 DNA，开发的 3-CHA 的检测限估计为 29.3 pM。在减少漏液信号的支持下，所实现的 3-CHA 显示出用于检测低浓度靶 DNA 的巨大潜力。
