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：
10645979
负责人：
Anna Schibel
金额：
$ 39.35万
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：
2023-08-11 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10645979
关键词：
Amino Acids Back Biological Biological Markers ChIP-seq Charge Complex Computer software DNA Diameter Disease Electrolytes Electronics Elements Enzymes Evaluation Exclusion Feasibility Studies Gene Expression Goals Homo Individual Length Ligation Lipid Bilayers Magnetism Measurement Measures Methodology Methods Monitor Motor Nucleic acid sequencing Peptide Sequence Determination Peptides Phase Positioning Attribute Post-Translational Protein Processing Preparation Protein translocation Proteins Proteome Proteomics RNA Reader Reading Frames Reproducibility Resolution Sampling Scheme Secondary Protein Structure Small Business Innovation Research Grant Stretching System Systems Integration Techniques Technology Time Variant War biological research data acquisition detection method high reward high risk improved instrumentation nanopore novel particle polypeptide prevent programs protein folding prototype research clinical testing sequencing platform single cell proteins single molecule synthetic peptide tool voltage

项目摘要

Project Summary In the field of proteomics, the ultimate goal is to determine the sequence and abundance of every protein variant in a sample. To reach this goal, we need techniques capable of single-molecule, high-accuracy, de novo protein sequencing. Unfortunately, current proteomics technologies lack the sensitivity, dynamic range, throughput, scale, and accuracy needed to meet this goal. During this project, we will develop technological components and methods required to enable state-of-the-art protein sequencing. More specifically, we will develop methods for controllably translocating a protein through a nanopore reader such that the protein can be sequenced with single amino acid resolution. Nanopore-based sequencing provides a path to characterizing both high- and low- abundance proteins by sequencing proteins one molecule at a time, but with the high accuracy, sensitivity and throughput needed to cover the wide dynamic range in protein abundance in the proteome. During this project, we will develop and build a novel prototype sequencing system, fully assess and optimize the associated workflow/methodology for highly accurate, reproducible, 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.

项目摘要在蛋白质组学领域，最终目标是确定每个蛋白质变体的顺序和丰度在样本中。为了实现这一目标，我们需要能够能够单分子，高精度和从头蛋白的技术测序。不幸的是，当前的蛋白质组学技术缺乏灵敏度，动态范围，吞吐量，规模和准确性才能实现这一目标。在此项目中，我们将开发技术组成部分和实现最先进的蛋白质测序所需的方法。更具体地说，我们将开发用于可以通过纳米孔读取器控制蛋白质，以便可以将蛋白质测序氨基酸分辨率。基于纳米孔的测序提供了表征高和低 - 的途径通过对蛋白进行测序一个分子，但具有高精度，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，我们将开发和建立一个新颖的原型测序系统，充分评估和优化相关的高度准确，可重现和多功能蛋白/肽表征的工作流/方法，以及演示各种蛋白质和肽的初始测序。在这个项目的结论下，我们将有成功证明了对实用纳米孔的蛋白质测序概念的可行性。