DNA nanopores are a recent class of bilayer-puncturing nanodevices that can help advance biosensing, synthetic biology, and nanofluidics. Here, we create archetypal lipid-anchored DNA nanopores and characterize them with a nanoprobe-based approach to gain essential information about their interactions with bilayers. The strategy determines the molecular accessibility of DNA pores with a nuclease and can thus distinguish between the nanopores’ membrane-adhering and membrane-spanning states. The analysis reveals, for example, that pores interact with bilayers in two steps whereby fast initial membrane tethering is followed by slower reorientation to the puncturing state. Tethering occurs for pores with one anchor, while puncturing requires multiple anchors. Both low and high-curvature membranes are good substrates for tethering, but efficient insertion proceeds only for high-curvature bilayers of the examined lipid composition. This is likely due to the localized lipid misalignments and the associated lower energetic barrier for pore permeation. Our study advances the fields of DNA nanotechnology and nanopores by overcoming the considerable experimental hurdle of efficient membrane insertion. It also provides mechanistic insights to aid the design of advanced nanopores, and offers a useful route to probe bilayer orientation of DNA nanostructures.

中文翻译：

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DNA纳米孔的定义的双层相互作用揭示了基于核酸酶的纳米探针策略。

DNA纳米孔是一类新型的双层穿刺纳米器件，可帮助推进生物传感，合成生物学和纳米流体技术。在这里，我们创建了原型脂质锚定的DNA纳米孔，并使用基于纳米探针的方法对其进行了表征，以获取有关其与双层相互作用的基本信息。该策略使用核酸酶确定DNA孔的分子可及性，因此可以区分纳米孔的膜粘附状态和跨膜状态。该分析表明，例如，毛孔在两个步骤中与双层相互作用，由此快速的初始膜束缚随后是较慢的重新定向至穿刺状态。带有一个锚的孔会发生系留，而刺孔则需要多个锚。低曲率膜和高曲率膜都是栓系的良好基质，但有效插入仅针对所检查脂质组合物的高曲率双层进行。这可能是由于局部脂质错位和相关的较低的能量渗透屏障所致。我们的研究克服了有效的膜插入的相当大的实验障碍，从而推进了DNA纳米技术和纳米孔的领域。它还提供了机械方面的见识，以帮助设计先进的纳米孔，并提供了探测DNA纳米结构的双层取向的有用途径。我们的研究克服了有效的膜插入的相当大的实验障碍，从而推进了DNA纳米技术和纳米孔的领域。它还提供了机械方面的见识，以帮助设计先进的纳米孔，并提供了探测DNA纳米结构的双层取向的有用途径。我们的研究克服了有效的膜插入的相当大的实验障碍，从而推进了DNA纳米技术和纳米孔的领域。它还提供了机械方面的见识，以帮助设计先进的纳米孔，并提供了探测DNA纳米结构的双层取向的有用途径。