Krishnan Swati, Ziegler Daniela, Arnaut Vera, Martin Thomas G, Kapsner Korbinian, Henneberg Katharina, Bausch Andreas R, Dietz Hendrik, Simmel Friedrich C
Physik-Department E14, Technische Universität München, Am Coulombwall 4a, 85748 Garching, Germany.
Zentrum für Nanotechnologie und Nanomaterialien/WSI, Technische Universität München, Am Coulombwall 4a, 85748 Garching, Germany.
Nat Commun. 2016 Sep 23;7:12787. doi: 10.1038/ncomms12787.
DNA-based nanopores are synthetic biomolecular membrane pores, whose geometry and chemical functionality can be tuned using the tools of DNA nanotechnology, making them promising molecular devices for applications in single-molecule biosensing and synthetic biology. Here we introduce a large DNA membrane channel with an ≈4 nm diameter pore, which has stable electrical properties and spontaneously inserts into flat lipid bilayer membranes. Membrane incorporation is facilitated by a large number of hydrophobic functionalizations or, alternatively, streptavidin linkages between biotinylated channels and lipids. The channel displays an Ohmic conductance of ≈3 nS, consistent with its size, and allows electrically driven translocation of single-stranded and double-stranded DNA analytes. Using confocal microscopy and a dye influx assay, we demonstrate the spontaneous formation of membrane pores in giant unilamellar vesicles. Pores can be created both in an outside-in and an inside-out configuration.
基于DNA的纳米孔是合成生物分子膜孔，其几何形状和化学功能可利用DNA纳米技术工具进行调节，使其成为单分子生物传感和合成生物学应用中很有前景的分子器件。在此，我们介绍一种直径约为4纳米的大型DNA膜通道，其具有稳定的电学性质，可自发插入平面脂质双分子层膜中。大量的疏水官能化作用，或者生物素化通道与脂质之间的链霉亲和素连接，有助于膜整合。该通道显示出约3纳安的欧姆电导，与其尺寸相符，并允许单链和双链DNA分析物进行电驱动转运。利用共聚焦显微镜和染料流入测定法，我们证明了在巨型单层囊泡中膜孔的自发形成。孔可以以由外而内和由内而外的构型形成。
