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

• 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
• 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

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.

项目概要 蛋白质组学领域的最终目标是单分子、高精度、从头蛋白质测序。 不幸的是，当前的蛋白质组学技术成本高昂，并且缺乏灵敏度、动态范围、通量、 实现这一目标所需的规模和准确性。在这个项目期间，我们将开始检查技术 实现最先进的蛋白质测序和表征所需的组件和方法。更多的 具体来说，我们将开发通过纳米孔读取器可控地转移蛋白质的方法，以便 可以对蛋白质进行测序。基于纳米孔的测序提供了一种表征高通量和低通量的途径 通过每次对一个分子进行蛋白质测序来检测低丰度蛋白质，但具有高精度、灵敏度和 覆盖蛋白质组中蛋白质丰度的广泛动态范围所需的通量。在这个项目期间， 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。我们将开发 构建新型蛋白质测序系统原型，充分评估和优化相关 用于高度控制和多功能蛋白质/肽表征的工作流程/方法，并展示初始 对各种蛋白质和肽进行测序。在这个项目结束时，我们将成功地 证明了基于纳米孔的实用蛋白质测序概念的可行性。