Conductance Fluctuations: A New Approach to Sequencing?

电导波动：测序的新方法？

• 批准号: 9922984
• 负责人: STUART LINDSAY
• 金额: $ 31.4万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9922984
• 关键词: Affect; Antibodies; Back; Binding; Biotin; Carbon Nanotubes; Charge; Chemicals; Chemistry; Clinic; Coupling; DNA; DNA Polymerase I; DNA-Directed DNA Polymerase; Data; Development; Devices; Dreams; Dyes; Electrodes; Electrons; Engineering; Epitopes; Evaluation; Feedback; Genome; Geometry; Goals; Government; Heterogeneity; Hour; Individual; Label; Measurement; Measures; Methods; Molecular; Molecular Conformation; Motion; Nature; Noise; Normal Statistical Distribution; Nucleotides; Peptides; Persons; Polymerase; Positioning Attribute; Process; Property; Proteins; Publishing; Running; Scanning Probe Microscopes; Scanning Tunneling Microscopy; Scheme; Semiconductors; Signal Transduction; Silicon; Speed; Streptavidin; Structure; Surface; Technology; Testing; Time; Transistors; Uncertainty; Variant; base; cost; design; enzyme activity; experimental study; genome sequencing; improved; novel strategies; polymerization; protein function; scale up; sequencing platform; single molecule; single walled carbon nanotube; solid state; voltage

Project Summary The objective of this two-year R21 project is to explore a very new approach to probing the dynamics of enzyme activity based on recently-discovered electronic properties of proteins. The long-term goal is to create a low-cost, high speed single-molecule genome sequencer with long reads, requiring no dyes or labels, with direct electronic readout. The technology has the potential for rapid reads (on the order of an hour per genome) using an integrated circuit chip. If this potential were to be fully realized, then use of genome sequencing in the clinic with near real-time feedback could become a reality. Initial data shows that high signal to noise measurement of protein fluctuations appears to be possible, and suggests that at least some of the fluctuations are associated with the function of the protein. However, these studies are limited by a great deal of variability owing to uncertainty about the chemical nature of contacts to the proteins. Therefore, our first aim is to engineer polymerases with built-in contacts, designed to self-assemble into a measurement device. We will test the construct with a scanning tunneling microscope, and, in the process, obtain design parameters for a sequencing device. Our second aim is to test at least one structure in a solid-state device, to check the validity of data obtained in a scanning tunneling microscopy. If successful, the data collected would lay the groundwork for a larger effort to scale up and improve accuracy to achieve a viable sequencing platform with the properties described above.

项目概要 这个为期两年的 R21 项目的目标是探索一种非常新的方法来探索动态 基于最近发现的蛋白质电子特性的酶活性。长期目标是创建 低成本、高速单分子基因组测序仪，具有长读长，无需染料或标签， 直接电子读数。该技术具有快速读取的潜力（大约每小时一小时） 基因组）使用集成电路芯片。如果要充分发挥这种潜力，那么利用基因组 具有近乎实时反馈的临床测序可能成为现实。初始数据显示高信号 蛋白质波动的噪声测量似乎是可能的，并且表明至少有一些 波动与蛋白质的功能有关。然而，这些研究受到很大的限制 由于与蛋白质接触的化学性质的不确定性而导致变异性。因此，我们的首要目标 是设计具有内置触点的聚合酶，旨在自组装成测量设备。我们 将使用扫描隧道显微镜测试该结构，并在此过程中获得设计参数 测序装置。我们的第二个目标是测试固态器件中的至少一种结构，以检查 扫描隧道显微镜中获得的数据的有效性。如果成功的话，收集到的数据将奠定 为更大程度地扩大规模和提高准确性奠定基础，以实现可行的测序平台 上述属性。