PFI-TT: A highly multiplexed readout system for single-molecule analysis

PFI-TT：用于单分子分析的高度多重读出系统

• 批准号: 2016555
• 负责人: Michael Roukes
• 金额: $ 25万
• 依托单位: California Institute of Technology
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-15 至 2023-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2016555&HistoricalAwards=false
• 关键词: PFI; TT; highly; multiplexed; readout

The broader impact/commercial potential of this Partnerships for Innovation - Technology Translation (PFI-TT) project is to develop instrumentation that will enable deep profiling of cellular proteins. The methods will be made broadly applicable in fundamental biological research. The project’s central goal is to develop and validate instrumentation for studying proteomics – the collection of proteins constituting the molecular machinery underlying all life forms. Genes, the molecular precursors of cellular machinery, are templates encoding how proteins are constructed within cells. The genomics revolution has recently enabled decoding of the human genome. Similar advances in proteomics technology has not occurred since genomic studies rely upon gene amplification to making billions of identical copies to enable analysis en masse. No similar molecular amplification process exists for proteins, hence single-molecule analyses are key. The proposed research relies upon the collaborative efforts of researchers spanning physics, engineering, chemistry, biology, and mathematics – from both academia and industry. Component technologies for a feasible, commercializable instrument will be pursued. This project’s highly collaborative and rich research environment provides an opportunity for the graduate student who will be involved in this effort to gain important career skills. The proposed project will develop instrumentation to unravel the staggering complexity of protein biology. An individual mammalian cell comprises roughly 3B proteins, spanning about 7,000 unique types, with concentrations spanning 8 orders of magnitude – ranging from cellular proteins present with just a few copies, to those with expression levels of order 100M. In human blood serum this range is a 1000X greater, spanning 11 orders. Resolving the full spectrum of protein constituents is essential to fundamental questions in biology and medicine. Detailed analyses – at the population level, yet with single-molecule resolution – will stratify fine details lost in existing consensus-type approaches to proteomics that “average over” the protein population, thus providing information only about the most prevalent species. Critical processes in health and disease are often determined by only a few copies of a protein molecule within a cell. Accordingly, single-molecule resolution of proteins and protein complexes in the presence of an immense overabundance of the most prevalent species is essential.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

该创新合作伙伴关系 - 技术转化 (PFI-TT) 项目的更广泛影响/商业潜力是开发能够对细胞蛋白质进行深入分析的仪器。该项目的中心目标是广泛适用于基础生物学研究。开发和验证用于研究蛋白质组学的仪器——构成所有生命形式的分子机制的蛋白质集合。基因是细胞机制的分子前体，是编码细胞内蛋白质如何构建的模板。人类基因组的解码尚未出现，因为基因组研究依赖基因扩增来产生数十亿相同的拷贝来进行大规模分析，因此单分子分析是关键。拟议的研究依赖于来自学术界和工业界的物理、工程、化学、生物学和数学研究人员的共同努力，该项目将提供高度协作和丰富的研究环境，以实现可行的、可商业化的仪器。该项目将为参与这项工作的研究生提供获得重要职业技能的机会，该项目将开发仪器来揭示蛋白质生物学的惊人复杂性，单个哺乳动物细胞包含大约 3B 种蛋白质，涵盖约 7,000 种独特的类型。浓度跨越 8 个数量级——从仅存在几个拷贝的细胞蛋白到表达水平为 100M 数量级的蛋白，在人类血清中，这个范围要大 1000 倍，跨越 11 个数量级。全谱蛋白质成分对于生物学和医学中的基本问题至关重要，在群体水平上进行单分子分辨率的详细分析将对现有共识型蛋白质组学方法中丢失的精细细节进行分层，这些方法对蛋白质进行“平均”。群体，因此仅提供有关最普遍物种的信息，通常仅由细胞内蛋白质分子的几个拷贝决定。大量过剩的最普遍的物种是必不可少的。该奖项反映了 NSF 的法定使命，并且通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。