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.

该创新技术转化伙伴关系（PFI-TT）项目的更广泛影响/商业潜力是开发能够深入分析细胞蛋白质的仪器。这些方法将广泛应用于基础生物学研究。该项目的中心目标是开发和验证用于研究蛋白质组学的仪器，蛋白质组学是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，是编码蛋白质如何在细胞内构建的模板。基因组学革命最近使人类基因组的解码成为可能。类似的蛋白质组学技术还没有出现，因为基因组研究依赖于基因扩增来制造数十亿个相同的拷贝来进行大规模分析。蛋白质没有类似的分子扩增过程，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学、工程学、化学、生物学和数学等领域的研究人员的合作努力。将寻求一种可行的、可商业化的仪器的组件技术。这个项目的高度协作和丰富的研究环境为研究生提供了一个机会，他们将参与这项工作，获得重要的职业技能。拟议中的项目将开发仪器来揭示蛋白质生物学惊人的复杂性。一个单独的哺乳动物细胞包含大约3B种蛋白质，跨越大约7000种独特的类型，浓度跨越8个数量级——从只有几个拷贝的细胞蛋白质到表达水平为100万数量级的细胞蛋白质。在人类血清中，这一范围要大1000倍，跨越11个目。解决蛋白质成分的全谱对生物学和医学的基本问题至关重要。详细的分析——在种群水平上，但使用单分子分辨率——将对现有的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“平均”蛋白质种群，从而仅提供有关最普遍物种的信息。健康和疾病的关键过程往往是由细胞内蛋白质分子的几个拷贝决定的。因此，在存在大量过剩的最普遍物种的情况下，蛋白质和蛋白质复合物的单分子分辨率是必不可少的。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。