Colorimetric detection of miRNA-21 by DNAzyme-coupled branched DNA constructs
Colorimetric detection of miRNA-21 by DNAzyme-coupled branched DNA constructs
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DOI：
10.1016/j.talanta.2020.120913
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发表时间：
2020-08-15
期刊：
影响因子：
6.1
通讯作者：
Pourghadamyari, Hossein
中科院分区：
文献类型：
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作者：
Hosseinzadeh, Elyas;Ravan, Hadi;Pourghadamyari, Hossein
核酸表达的波动以及对这些表达变化的超灵敏和特异性检测是分子医学和临床治疗诊断学的重要课题。本研究报道了一种新型 DNAzyme 偶联支链杂交链式反应 (b-HCR) 测定法，用于有效信号放大检测 miRNA。该测定由与 miR-21 杂交的翻译器 (T) 组成，通过发夹 1 (H-1) 和发夹 2 (H-2) 启动第一个 HCR。初级 HCR 为通过发夹 3 (H-3) 和发夹 4 (H-4) 组装出芽的众多分支提供主链。在存在氯化血红素的情况下，嵌入 H1 和 H4 中的富含 G 的结构域在暴露于靶标时会产生活性 G-四链体脱氧核糖核酸酶，该靶标可以催化无色底物氧化为有色产物。本方法有潜力用于定量检测 miR-21，灵敏度和动态范围分别为 1 pM 和 1 pM 至 1 nM。
Fluctuation of nucleic acid expression and ultrasensitive and specific detection of these variations in expression is a crucial subject in molecular medicine and clinical theranostics. A novel DNAzyme-coupled branched hybridization chain reaction (b-HCR) assay is reported for efficient signal-amplified detection of miRNA in this study. This assay was composed of a translator (T) hybridized with miR-21 to initiate the first HCR by hairpin 1 (H-1) and hairpin 2 (H-2). The primary HCR provided a backbone chain for numerous branches budding through hairpin 3 (H-3) and hairpin 4 (H-4) assembles. In the presence of hemin, the G-rich domains embedded in H1 and H4 produce an active G-quadruplex DNAzyme upon exposure to a target that could catalyze the oxidation of colorless substrate to colored product. The present approach has the potential to be used for quantitative detection of miR-21 with a sensitivity and a dynamic range of 1 pM and 1 pM to 1 nM, respectively.