Controlled Protein Translocation in Nanopores for Sequencing Applications

用于测序应用的纳米孔中的受控蛋白质易位

基本信息

批准号：
10574679
负责人：
Eric Mayo Peterson
金额：
$ 25.07万
依托单位：
ELECTRONIC BIOSCIENCES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10574679
关键词：
Amino Acids Back Biological ChIP-seq Complex Computer software DNA Diameter Disease Electronics Elements Enzymes Evaluation Exclusion Feasibility Studies Gene Expression Goals Homo Individual Length Ligation Lipid Bilayers Magnetism Measurement Methodology Methods Monitor Motor Nucleic acid sequencing Peptide Sequence Determination Peptides Positioning Attribute Post-Translational Protein Processing Preparation Protein translocation Proteins Proteome Proteomics RNA Reader Reading Frames Sampling Scheme Secondary Protein Structure Stretching System Systems Integration Technology Time Variant War biological research biomarker discovery biomarker validation cost data acquisition high reward high risk improved instrumentation nanopore novel particle prevent programs protein folding prototype research clinical testing sequencing platform single cell proteins single molecule synthetic peptide voltage

项目摘要

Project Summary The ultimate goal in the field of proteomics is single-molecule, high-accuracy, de novo protein sequencing. Unfortunately, current proteomics technologies are costly, and lack the sensitivity, dynamic range, throughput, scale, and accuracy needed to meet this goal. During this project, we will begin examining the technological components and methods required to enable state-of-the-art protein sequencing and characterization. More specifically, we will develop methods for controllably translocating a protein through a nanopore reader such that the protein can be sequenced. Nanopore-based sequencing provides a path to characterizing both high- and low-abundance proteins by sequencing proteins one molecule at a time, but with high accuracy, sensitivity and throughput needed to cover the wide dynamic range in protein abundance in the proteome. During this project, we will develop a completely new nanopore-based technology for de novo protein sequencing. We will develop and build a novel protein sequencing system prototype, fully assess and optimize the associated workflow/methodology for highly controlled and versatile protein/peptide characterization, and demonstrate initial sequencing for various proteins and peptides. At the conclusion of this project, we will have successfully demonstrated feasibility for a practical nanopore-based protein sequencing concept.

项目摘要蛋白质组学领域的最终目标是单分子，高临界性，从头蛋白质测序。不幸的是，当前的蛋白质组学技术是昂贵的，并且缺乏灵敏度，动态范围，吞吐量，规模和准确性才能实现这一目标。在这个项目中，我们将开始研究技术启用最先进的蛋白质测序和表征所需的组件和方法。更多的具体而言，我们将开发通过纳米孔读取器控制蛋白质的方法，这样可以测序蛋白质。基于纳米孔的测序提供了表征高和表征的途径通过对蛋白进行测序一个分子，但具有很高的精度，灵敏度和覆盖蛋白质蛋白质丰度的广泛动态范围所需的吞吐量。在这个项目中，我们将开发一种全新的基于纳米孔的技术，用于从头蛋白质测序。我们将发展并建立一个新型的蛋白质测序系统原型，充分评估和优化相关的高度控制和多功能蛋白/肽表征的工作流/方法，并证明初始对各种蛋白质和肽进行测序。在这个项目的结论中，我们将成功证明了实用的基于纳米孔的蛋白质测序概念的可行性。