BRIGE: Fluorescence Based Single Molecule Force Spectroscopy with DNA Nanotechnology

BRIGE：基于荧光的单分子力光谱与 DNA 纳米技术

• 批准号: 1228104
• 负责人: Carlos Castro
• 金额: $ 17.46万
• 依托单位: Ohio State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-15 至 2014-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1228104&HistoricalAwards=false
• 关键词: BRIGE; Fluorescence; Based; Single; Molecule

This Broadening Participation Research Initiation Grant in Engineering (BRIGE) provides funding for the development of a nanoscale device for fluorescence-based high throughput single molecule force spectroscopy. The device will be assembled using the recently developed nanotechnology DNA origami. DNA origami enables the construction of nanoscale objects with unprecedented geometric complexity via programmed molecular self-assembly. The device will comprise a stiff framework of bundles of double-stranded DNA, attachment points for two biomolecules, a flexible polymer force probe, and fluorescent molecules to act as a readout of the interaction between the two biomolecules of interest. The force probe will facilitate binding between the biomolecules, and subsequently apply a known force acting to rupture the interaction. The bond lifetime will be monitored using a fluorescence readout, which is amenable to high throughput data collection. Interaction lifetimes will be measured as a function of force to determine kinetic parameters that govern the molecular interaction. Initial proof-of-principle experiments will probe DNA and RNA base-pairing interactions. Ultimately, the device will be implemented to study protein-DNA, protein-RNA, and protein-protein systems.Single molecule force spectroscopy studies have provided critical insight into the interactions that stabilize biomolecular machinery and regulate the function of cells. However, the widespread application of force spectroscopy has been hindered by the need for cumbersome and expensive equipment and low throughput data acquisition. If successful, this research will enable economical and high-throughput force spectroscopy studies of biomolecular machinery. The ultimate goal is to develop a device that can be packaged and distributed for widespread application on basic laboratory fluorescence microscopes. This award will also support the establishment of a biomolecular machinery short course to be offered to high school age students at summer engineering camps and the development of a research seminar course aimed to recruit underrepresented engineering students to pursue graduate level research.

这个扩大参与工程研究启动补助金（BRIGE）为基于荧光的高通量单分子力光谱的纳米级设备的开发提供资金。 该装置将使用最近开发的纳米技术DNA折纸组装。 DNA“折纸”通过程序化的分子自组装，能够构建具有前所未有的几何复杂性的纳米级物体。 该装置将包括双链DNA束的刚性框架、两个生物分子的附着点、柔性聚合物力探针和荧光分子，以用作感兴趣的两个生物分子之间的相互作用的读出。 力探针将促进生物分子之间的结合，并随后施加已知的力以破坏相互作用。 将使用荧光读数监测粘合剂寿命，该荧光读数适合于高通量数据收集。 将测量相互作用寿命作为力的函数，以确定控制分子相互作用的动力学参数。 最初的原理验证实验将探测DNA和RNA碱基配对的相互作用。 最终，该装置将被用于研究蛋白质-DNA、蛋白质-RNA和蛋白质-蛋白质系统。单分子力谱研究为稳定生物分子机制和调节细胞功能的相互作用提供了重要的见解。 然而，力谱的广泛应用一直受到阻碍的笨重和昂贵的设备和低吞吐量的数据采集的需要。 如果成功，这项研究将实现对生物分子机械的经济且高通量的力谱研究。 最终目标是开发一种可以包装和分发的设备，用于基本实验室荧光显微镜的广泛应用。 该奖项还将支持建立一个生物分子机械短期课程，提供给高中年龄的学生在夏令营工程和研究研讨会课程的发展，旨在招募代表性不足的工程专业学生进行研究生水平的研究。