Single-Molecule Mechanochemical Sensing for Multiplexed Tasks: A Topochemical Strategy

本提案的研究目标是采用多学科方法创新具有超高灵敏度的生物传感策略，以检测多种化学或生物靶点。检测将通过监测在结合化学或生化目标时传感模板的张力或长度的变化来完成。为了同时检测多个目标，这是下一代生物传感器的基本特征，研究小组将在一个模板中集成不同的传感单元。每个传感单元被设计为报告特定目标的结合。该项目的成功完成将提供新的生物传感范例，与传统生物传感器相比，将单分子生物传感器转化为具有高灵敏度和优越效率的可行设备具有巨大潜力。在这个项目中，研究者计划将教育元素融入到研究项目中。研究者的实验室将通过招募高中生和大学生来进行这项由国家科学基金会资助的研究。研究是模块化设计，以促进这些学生参与到项目中来。此外，研究小组还将通过帮助K-12学生进行科学活动来接触公众。该项目的智力优势在于使用单个DNA分子作为传感模板，而分析物的检测基于机械化学原理。当单个化学或生化靶点与单分子DNA模板发生化学结合时，模板的力学性能，如拉伸力和/或延伸性将发生变化。为了检测这种在皮牛顿量级和纳米量级的微小信号变化，将使用单分子技术，如光镊和磁镊。由于分析物检测是在单分子水平上实现的，因此不需要化学扩增，这增加了机械化学传感中材料使用的效率。为了提高浓度检测的极限，许多传感单元将被纳入模板使用分子生物学方法，如滚动圈扩增（RCA）。此外，信号被局部传感单元的空间排列放大，这一特征遵循拓扑化学原理。为了提高传感的吞吐量，将使用磁镊子，以便数百个单分子传感模板可以固定在表面上并同时监测。拓扑化学传感策略有望导致可行的高灵敏度和高效率的单分子生物传感器。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The research objective of this proposal is to take a multidisciplinary approach to innovate biosensing strategies with ultrahigh sensitivity to detect multiple chemical or biological targets. The detection will be accomplished by monitoring the variation of either tension or length of the sensing template upon binding of chemical or biochemical targets. To detect many targets simultaneously, which is an essential feature in next generation biosensors, the research team will incorporate different sensing units in a single template. Each sensing unit is designed to report the binding of specific targets. Successful completion of this project will provide new biosensing paradigms with great potential to transform single-molecule biosensors into viable devices with high sensitivity and superior efficiency compared to conventional biosensors. During this project, the investigator plans to integrate educational elements into research projects. The investigator's lab will train high-school and college students by recruiting them to carry out this NSF funded research. Research is modularly designed to facilitate participation of these students into the project. In addition, the research team will reach out to the public by helping K-12 students for scientific activities.The intellectual merit of this project lies in the employment of single DNA molecules as sensing templates while detection of analytes is based on mechanochemical principles. Upon chemical binding of individual chemical or biochemical targets to the single-molecule DNA template, the mechanical property of the template such as its tensile force and/or extension will change. To detect such a tiny signal change on the order of picoNewton in force and nanometer in extension, single-molecule techniques such as optical tweezers and magnetic tweezers will be used. Since analyte detection is achieved at the single-molecule level, chemical amplification is not necessary, which increases the efficiency of materials usage in the mechanochemical sensing. To improve concentration detection limit, many sensing units will be incorporated in the template using molecular biology methods such as Rolling Circle Amplification (RCA). In addition, signal is amplified by spatial arrangement of localized sensing units, a feature that follows topochemical principles. To increase the throughput of the sensing, magnetic tweezers will be used so that hundreds of single-molecule sensing templates can be immobilized on a surface and monitored simultaneously. The topo chemical sensing strategies are anticipated to lead to viable single-molecule biosensors with high sensitivity and efficiency.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.