RNA Nanomotor Based Active Devices for Biology and Medicine

描述(申请人提供)：纳米技术正在彻底改变我们制造和设计小型、廉价和超灵敏有源设备和系统的能力。生产用于快速和并行检测或健康监测的小型医疗设备的需求正在加剧。此外，随着材料科学和工程领域的每一次新发展，微制造器件变得越来越小，设计和制造生物启发的纳米器件和传感器以及超紧凑型电源来驱动这些器件和传感器的需求正在涌现。大自然可以提供工具来满足上述需求。分子马达，如ATPase[Noji等人，1997]、细菌鞭毛[Sowa等人，2005]或病毒DNA包装马达[Guo，2002；Shu等人，2003]可以用来在纳米尺度上合成能量。其中一些马达可以产生高达数十或数百皮牛顿的力。它们中的一些可以利用能量并通过ATP水解产生力量，或者通过电动机动力或pH梯度以ATP的形式利用力量并产生能量，效率在80%-100%的范围内[Yasuda等人，2001；Aksimentiev等人，2004]。其中一些电机的转速可以达到每分钟100-1000转[Sowa等人，2005]。近年来，纳米制造能力已经发展到可以在硅片上的特定位置生长20 nm以下的纳米孔、纳米线和纳米管的程度，从而这些结构可能与生物马达对接，用于纳米机械、过滤、移动以及能量产生和采集等应用。在这个项目中，我们计划开发基于生物纳米电机的活性纳米结构和系统。我们这里的重点将是使用噬菌体phi29 DNA包装纳米马达，它由称为包装RNA或“PRNA”的小RNA分子驱动和驱动。这种纳米马达已经被证明在将phi29基因组DNA运送到原衣壳动物中起到了新的和重要的作用。随着人们对这些新体系的结构和机制的理解有了更多的进展，现在是时候使用基于生物纳米技术的方法来评估这些结构，并使用自上而下和自下而上的制造技术来探索这些纳米电机和合成结构之间的界面，以形成活性纳米结构和纳米系统。我们的核心平台将由PRNA驱动马达组成，这些马达通过二维自组装DNA晶体固定在微机械硅或氧化铝基膜上的纳米孔膜上。DNA自组装层的使用将确保纳米电机的完整性和功能性。这一基本平台的发展和特点本身就是一项重大挑战，需要采取协调一致的跨学科方法。我们将把纳米电机集成在一个基于混合硅基的设备中并演示其操作，然后集成没有衣壳的纳米电机并演示dsDNA通过电机的移位。一旦这些任务完成，就有可能研究各种技术模块，如微流控通道内的主动泵送表面、主动筛分和过滤，以及许多其他与生物和医学直接相关的应用。

DESCRIPTION (provided by applicant): Nanotechnology is truly revolutionizing our ability to manufacture and design active devices and systems that are small, cheap, and ultra-sensitive. The need to produce small medical devices for rapid and parallel detection or health monitoring is intensifying. Moreover, as the micro-fabricated devices get smaller and smaller with each new development in materials science and engineering, the need to design and fabricate biologically inspired nanoscale devices and sensors and ultra-compact power sources to drive these devices and sensors are emerging. Nature can provide the tools to address the above needs. Molecular motors, such as ATPase [Noji et al., 1997], bacterial flagellar [Sowa et al., 2005], or viral DNA packaging motors [Guo, 2002; Shu et al., 2003] can be utilized to synthesize power at the nano-scale. Some of these motors can generate force up to tens or hundreds of pico Newtons. Some of them can use energy and generate force, via ATP hydrolysis, or use the force and generate energy in the form of ATP, via electrolocomotive force or pH gradients, with efficiencies in the range of 80-100% [Yasuda et al., 2001; Aksimentiev et al., 2004]. Some of these motors can have rotational speeds of 100-1000 rpm [Sowa et al., 2005]. In the recent years, nanofabrication capabilities have progressed to a point where sub 20nm nanopores, nanowires, and nanotubes can be grown at specific locations on a silicon wafer such that these structures can possibly be interfaced with biological motors for applications such as nanomechanics, filtration, locomotion, and energy generation and harvesting. In this project, we propose to develop active nanostructures and systems based on biological nanomotors. Our focus here would be the use of the bacteriophage phi29 DNA packaging nanomotor that is driven and geared by small RNA molecules termed packaging RNA or "pRNA". This nanomotor has been shown to play a novel and essential role in transporting phi29 genomic DNA into procapsids. As more progress is made in understanding the structure and mechanisms of these novel systems, it is time to evaluate these structures using bionanotechnology-based approaches and to explore the interface between these nanomotors and synthetic structures using top down and bottoms up fabrication technology to form active nanostructures and nanosystems. Our core platform will consist of the pRNA-driving motors anchored on nanoporous membranes on micromachined silicon or Alumina based membrane via a 2-dimensional self- assembled DNA crystal. The use of the DNA self assembled layer will ensure the integrity and functionality of the nanomotor. The development and characterization of this basic platform is a significant challenge in itself and requires a cohesive interdisciplinary approach. We will integrate the nanomotor in a hybrid silicon based device and demonstrate its operation, and then integrate the nanomotor without the capsid and demonstrate the translocation of dsDNA through the motor. Once these tasks are accomplished, it will be possible to investigate various technology modules such as active pumping surfaces within microfluidic channels, active sieving and filtration, and many other applications directly relevant to biology and medicine.