A large size-selective DNA nanopore with sensing applications
A large size-selective DNA nanopore with sensing applications
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DOI：
10.1038/s41467-019-13284-1
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发表时间：
2019-12
影响因子：
16.6
通讯作者：
Rasmus P. Thomsen;M. Malle;A. Okholm;Swati Krishnan;S. Bohr;R. S. Sørensen;O. Ries;S. Vogel;F. Simmel;N. Hatzakis;J. Kjems
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文献类型：
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作者：
Rasmus P. Thomsen;M. Malle;A. Okholm;Swati Krishnan;S. Bohr;R. S. Sørensen;O. Ries;S. Vogel;F. Simmel;N. Hatzakis;J. Kjems
跨膜纳米结构如离子通道和转运蛋白通过控制分子在脂质双层中的流动来执行关键的生物功能。许多工作已经进入工程人工纳米孔，并在分子的选择性门控，生物分子的无标记检测/传感和DNA测序中的应用已经显示出希望。在这里，我们使用DNA折纸来创建一个合成的9 nm宽的DNA纳米孔，由可编程的脂化皮瓣控制，并配备了大小选择性门控系统，用于大分子的易位。通过透射电子显微镜（TEM）验证纳米孔到脂质双层中的成功组装和插入，同时使用光学方法（包括单分子、全内反射荧光（TIRF）显微镜）研究货物的选择性易位和孔机械敏感性。尺寸特定的货物转运和阿托西肽触发的孔的开放被证明，表明DNA纳米孔可以作为外部信号的实时检测系统，为各种高度并行化的传感应用提供潜力。
Transmembrane nanostructures like ion channels and transporters perform key biological functions by controlling flow of molecules across lipid bilayers. Much work has gone into engineering artificial nanopores and applications in selective gating of molecules, label-free detection/sensing of biomolecules and DNA sequencing have shown promise. Here, we use DNA origami to create a synthetic 9 nm wide DNA nanopore, controlled by programmable, lipidated flaps and equipped with a size-selective gating system for the translocation of macromolecules. Successful assembly and insertion of the nanopore into lipid bilayers are validated by transmission electron microscopy (TEM), while selective translocation of cargo and the pore mechanosensitivity are studied using optical methods, including single-molecule, total internal reflection fluorescence (TIRF) microscopy. Size-specific cargo translocation and oligonucleotide-triggered opening of the pore are demonstrated showing that the DNA nanopore can function as a real-time detection system for external signals, offering potential for a variety of highly parallelized sensing applications.