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Sequencing by Direct Electrical Measurements of Polymerase Fluctuations

通过聚合酶波动的直接电测量进行测序

基本信息

批准号：
10359183
负责人：
STUART LINDSAY
金额：
$ 70.59万
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：
2020
资助国家：
美国
起止时间：
2020-05-15 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10359183
关键词：
Agreement Binding Biochemistry Cells ChIP-seq Characteristics Charge Clinic DNA Sequence DNA sequencing DNA-Directed DNA Polymerase Data Devices Dyes Electrodes Electron Transport Electronics Engineering Evaluation Event Feedback Gel Genome Geometry Goals Hour Individual Kinetics Label Ligands Liquid substance Measurement Measures Methylation Molecular Conformation Nucleotides Oxidation-Reduction Periodicity Polymerase Post-Translational Protein Processing Problem Solving Proteins Reporting Resolution Role Running Scanning Probe Microscopes Signal Transduction Speed Streptavidin Technology Testing Time Variant base chemical kinetics cost data acquisition design detector electrical measurement experimental study genome sequencing improved integrated circuit metallicity molecular dynamics nanofabrication nanoimprinting new technology polymerization prototype single molecule solid state temporal measurement tool

项目摘要

PROJECT SUMMARY Our 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 based on simple two terminal devices. 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. Our data shows that large-amplitude polymerase fluctuations are associated with polymerase activity. Our first goal is to identify signals associated with each nucleotide incorporation with high accuracy. This would enable sequencing by means of cyclic addition of nucleotides, but with the advantage that homopolymer runs of sequence would be counted directly. Our second aim is to identify the individual nucleotides being incorporated based on the details of the signals generated at each incorporation. This would allow a sequencer to run at the free-running speed of the polymerase in the presence of all four nucleotidetriphosphates, so that a wafer of 10,000 devices could produce a genome's worth of reads in an hour. Our third aim is to develop scalable technology for fabrication of prototype solid-state devices.

项目摘要我们的目标是建立一个低成本，高速度的单分子基因组测序仪，具有长的读取，不需要染料或标记，具有直接电子读出。该技术具有快速读取的潜力（大约每个基因组一小时）。如果这种潜力是要完全实现，那么在临床上使用基因组测序并提供近实时反馈可能会成为一种现实我们的数据表明，大幅度的聚合酶波动与聚合酶活性。我们的第一个目标是以高精度识别与每个核苷酸掺入相关的信号。这将能够通过核苷酸的环状添加进行测序，但具有以下优点：序列将被直接计数。我们的第二个目标是鉴定被掺入的单个核苷酸基于在每次掺入时产生的信号的细节。这将允许测序仪在在所有四种核苷酸磷酸存在下聚合酶的自由运行速度，使得 1万台设备可以在一小时内产生一个基因组的读数。我们的第三个目标是开发可扩展的制造原型固态器件的技术。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Heavy Water Reduces the Electronic Conductance of Protein Wires via Deuteron Interactions with Aromatic Residues.

DOI：
10.1021/acs.nanolett.3c02263
发表时间：
2023-09
期刊：
Nano letters
影响因子：
10.8
作者：
Sepideh Afsari;Sohini Mukherjee;Nicholas Halloran;Giovanna Ghirlanda;Eathen O Ryan;Xu Wang;S. Lindsay
通讯作者：
Sepideh Afsari;Sohini Mukherjee;Nicholas Halloran;Giovanna Ghirlanda;Eathen O Ryan;Xu Wang;S. Lindsay

Electronic Transport in Molecular Wires of Precisely Controlled Length Built from Modular Proteins.