Folding-upon-binding and signal-on electrochemical DNA sensor with high affinity and specificity.

Folding-upon-binding and signal-on electrochemical DNA sensor with high affinity and specificity.
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DOI:
10.1021/ac501418g
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发表时间:
2014-09-16
影响因子:
7.4
通讯作者:
Ricci, Francesco
Ricci, Francesco
中科院分区:
化学1区
文献类型:
--
作者:
Idili, Andrea;Amodio, Alessia;Vidonis, Marco;Feinberg-Somerson, Jacob;Castronovo, Matteo;Ricci, Francesco

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在这里,我们研究了一种新的信号上的电化学DNA传感器的基础上使用的钳形DNA探针,结合互补的目标序列,通过两个不同的和顺序的事件,这导致形成的三链体DNA结构。我们证明,这种目标结合机制可以提高识别的亲和力和特异性,而不是经典的探针仅基于沃森-克里克识别。通过使用电化学信号报告的构象变化,我们证明了一个信号上的E-DNA传感器与目标结合时的信号增益高达400%。此外,我们能够以纳摩尔亲和力检测到短至10个碱基(KD = 0.39 nM)的完全匹配的靶标。最后,由于分子的“双重检查”所提供的伴随沃森-克里克和Hoogsteen碱基配对参与目标识别,我们的传感器提供了良好的歧视效率对单碱基错配的目标。
Here we investigate a novel signal-on electrochemical DNA sensor based on the use of a clamp-like DNA probe that binds a complementary target sequence through two distinct and sequential events, which lead to the formation of a triplex DNA structure. We demonstrate that this target-binding mechanism can improve both the affinity and specificity of recognition as opposed to classic probes solely based on Watson–Crick recognition. By using electrochemical signaling to report the conformational change, we demonstrate a signal-on E-DNA sensor with up to 400% signal gain upon target binding. Moreover, we were able to detect with nanomolar affinity a perfectly matched target as short as 10 bases (KD = 0.39 nM). Finally, thanks to the molecular “double-check” provided by the concomitant Watson–Crick and Hoogsteen base pairings involved in target recognition, our sensor provides excellent discrimination efficiency toward a single-base mismatched target.
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