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A proof of concept application of aptachain: ligand-induced self-assembly of a DNA aptamer

机译：Aptachain的概念应用证明：配体诱导的DNA适体的自组装

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摘要

A challenge for the use of aptamers as biosensors is how to signal the occurrence of their ligand binding event into a signal that can be exploited in a detection scheme. Here, we present the concept of "aptachain" formation, where an aptamer is split into two overlapping or staggered strands and assembles into an extended oligomer upon ligand binding. This assembly of aptamers can then be used as a way to detect ligand binding by the aptamer. As an example of this concept, we employed the cocaine-binding aptamer as a model system, used its ability to tightly bind quinine and demonstrated its capability in a gold nanoparticle-based biosensing application. We used isothermal titration calorimetry to demonstrate that, when split into two overlapping DNA strands, the aptamer remains functional. Size-exclusion chromatography showed that the quinine-bound oligos form a larger assembly of aptamer units than in the absence of ligand. Finally, we used the oligomer forming ability of the aptachain oligos in a biosensor application for quinine that brings gold nanoparticles closer together resulting in a shift in their plasmonic resonance to a longer wavelength and an observed colour shift. We propose that splitting aptamers into overlapping strands that form oligomers in the presence of a ligand, aptachain formation, will be generally applicable to aptamers and prove useful in a variety of biotechnology applications.
机译：作为生物传感器使用适体的挑战是如何将它们的配体绑定事件的发生信号以在检测方案中可以利用的信号中发出信号。这里，我们介绍了“Aptachain”的概念，其中适体被分成两个重叠或交错的股线，并在配体结合时组合成延长的低聚物。然后，这种适体的组装可以用作检测APTamer的配体结合的方法。作为这种概念的一个例子，我们使用可卡因结合适体作为模型系统，使用其紧紧结合奎宁的能力并证明了其基于金纳米粒子的生物传感应用的能力。我们使用等温滴定热量测定法证明，当分成两个重叠的DNA链时，适体仍然是功能性的。尺寸排阻色谱显示奎宁 - 结合的寡核苷酸形成较大的适体单元组装，而不是在没有配体的情况下。最后，我们使用了Aptachain寡核苷酸在生物传感器应用中的低聚物形成能力，所述奎宁净化在一起，使得较近的金纳米颗粒较近，从而使其等离子体共振的变化与较长波长和观察到的颜色移位。我们提出将适体分裂成重叠的链，其在配体存在下形成低聚物，Aptachain形成，通常适用于适体并在各种生物技术应用中证明是可用的。

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