Li Hui, Wang Shaoying, Ji Zhouxiang, Xu Congcong, Shlyakhtenko Lyudmila S, Guo Peixuan
Center for RNA Nanobiotechnology and Nanomedicine; Division of Pharmaceutics and Pharmaceutical Chemistry, College of Pharmacy; Department of Physiology & Cell Biology, College of Medicine; Dorothy M. Davis Heart and Lung Research Institute and James Comprehensive Cancer Center, The Ohio State University, Columbus, OH, 43210, USA.
UNMC Nanoimaging Core Facility, Department of Pharmaceutical Sciences, College of Pharmacy University of Nebraska Medical Center, Omaha, NE, 68182, USA.
Nano Res. 2019 Aug;12(8):1952-1958. doi: 10.1007/s12274-019-2463-z. Epub 2019 Jul 11.
Nanotubes are miniature materials with significant potential applications in nanotechnological, medical, biological and material sciences. The quest for manufacturing methods of nano-mechanical modules is in progress. For example, the application of carbon nanotubes has been extensively investigated due to the precise width control, but the precise length control remains challenging. Here we report two approaches for the one-pot self-assembly of RNA nanotubes. For the first approach, six RNA strands were used to assemble the nanotube by forming a 11 nm long hollow channel with the inner diameter of 1.7 nm and the outside diameter of 6.3 nm. For the second approach, six RNA strands were designed to hybridize with their neighboring strands by complementary base pairing and formed a nanotube with a six-helix hollow channel similar to the nanotube assembled by the first approach. The fabricated RNA nanotubes were characterized by gel electrophoresis and atomic force microscopy (AFM), confirming the formation of nanotube-shaped RNA nanostructures. Cholesterol molecules were introduced into RNA nanotubes to facilitate their incorporation into lipid bilayer. Incubation of RNA nanotube complex with the free-standing lipid bilayer membrane under applied voltage led to discrete current signatures. Addition of peptides into the sensing chamber revealed discrete steps of current blockage. Polyarginine peptides with different lengths can be detected by current signatures, suggesting that the RNA-cholesterol complex holds the promise of achieving single molecule sensing of peptides.
纳米管是一种微型材料,在纳米技术、医学、生物学和材料科学领域具有重要的潜在应用价值。目前正在探索纳米机械模块的制造方法。例如,由于能够精确控制宽度,碳纳米管的应用已得到广泛研究,但精确控制长度仍然具有挑战性。在此,我们报告了两种用于RNA纳米管一锅法自组装的方法。对于第一种方法,使用六条RNA链通过形成一个内径为1.7 nm、外径为6.3 nm、长11 nm的中空通道来组装纳米管。对于第二种方法,设计六条RNA链通过互补碱基配对与相邻链杂交,形成一个具有六螺旋中空通道的纳米管,类似于第一种方法组装的纳米管。通过凝胶电泳和原子力显微镜(AFM)对制备的RNA纳米管进行了表征,证实了纳米管形状的RNA纳米结构的形成。将胆固醇分子引入RNA纳米管以促进其融入脂质双层。在施加电压的情况下,将RNA纳米管复合物与独立的脂质双层膜孵育会产生离散的电流信号。向传感腔中添加肽会显示出离散的电流阻断步骤。不同长度的聚精氨酸肽可以通过电流信号进行检测,这表明RNA-胆固醇复合物有望实现对肽的单分子传感。