Conductance Fluctuations: A New Approach to Sequencing?

电导波动：测序的新方法？

基本信息

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
项目类别：
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

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

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项目摘要

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.

项目摘要这个为期两年的R21项目的目的是探索一种非常新的方法来探测动态基于蛋白质最近发现的电子特性的酶活性。长期目标是创建低成本的高速单分子基因组测序仪长读数，不需要染料或标签，直接电子读数。该技术具有快速阅读的潜力（按照一个小时的顺序基因组）使用集成电路芯片。如果要充分实现这种潜力，则使用基因组在诊所中进行接近实时反馈的测序可能会成为现实。初始数据显示高信号对于蛋白质波动的噪声测量似乎是可能的，并表明至少一些波动与蛋白质的功能有关。但是，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，可变性。因此，我们的第一个目标用于具有内置触点的工程聚合酶，旨在自组装到测量设备中。我们将使用扫描隧道显微镜测试结构，并在此过程中获得设计参数测序设备。我们的第二个目标是在固态设备中至少测试一个结构，以检查在扫描隧道显微镜中获得的数据的有效性。如果成功，收集的数据将放置基础工作，以进行更大的努力，以扩大和提高准确性，以实现使用可行的测序平台上述属性。