CAREER: Imaging the Chemical Nature of Intracellular Phase Separation by Functional Stimulated Raman Scattering Spectro-microscopy

职业：通过功能受激拉曼散射光谱显微镜对细胞内相分离的化学性质进行成像

• 批准号: 2240092
• 负责人: Lu Wei
• 金额: $ 62.5万
• 依托单位: California Institute of Technology
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2240092&HistoricalAwards=false
• 关键词: CAREER; Imaging; Chemical; Nature; Intracellular

With support from the Chemical Measurement and Imaging (CMI) Program, and co-funding from the Chemistry of Life Processes (CLP) Program and the Chemical Structure, Dynamics, and Mechanisms-B (CSDM-B) Program in the CHE Division and the Molecular Biophysics Cluster in the MCB Division, Lu Wei and her research team at the California Institute of Technology are developing new optical imaging strategies that aim to improve our ability to study intracellular phenomena such as those relevant to neurodegenerative diseases. The research will result in new instrumentation, new imaging probes, and new labeling methods, all targeting new chemical insights into the behavior of proteins and RNA in live-cell settings. The highly interdisciplinary program will provide research opportunities for graduate and undergraduate students. Outreach efforts in the Wei group are planned to provide exposure to science to both K-12 and undergraduate students.The formation of liquid-liquid phase separation (LLPS) and its role in biological function has been extensively studied with many exciting new observations and discoveries. However, there is much yet to be learned especially regarding the biophysical and biochemical principles underlying the fundamental interactions among biomacromolecules, particularly protein and RNA and their ability to form condensates. To investigate key intracellular phase-separated condensates in live cells, the Wei lab is developing new stimulated Raman spectral-imaging strategies that are noninvasive and quantitative, and offer high spatial and temporal resolution. Specifically, the research program aims to (1) develop functional vibrational probes and labeling approaches to quantify the composition and sense the intra-condensate interactions from the identified LLPS systems for both in vitro and live-cell interrogations; (2) build new instrumentation modules to achieve up to 20-fold sensitivity enhancement for chemical mapping in live cells; and (3) perform in-depth live-cell investigations with advanced data analysis to piece together unknown chemical features and interaction principles in native and complex cellular settings.This award is co-funded by the CMI, CLP and CSDM-B Programs in the Division of Chemistry in the Directorate for Mathematical and Physical Sciences (MPS) and by the Molecular Biophysics Program in the Division of Molecular and Cellular Biosciences (MCB) in the Directorate for Biological Sciences (BIO).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.

在化学测量和成像(CMI)计划的支持下，在CHE部门的生命过程化学(CLP)计划和化学结构、动力学和机制B(CSDM-B)计划以及MCB部门的分子生物物理学集群的共同资助下，加州理工学院的卢伟和她的研究团队正在开发新的光学成像策略，旨在提高我们研究细胞内现象(如与神经退行性疾病相关的现象)的能力。这项研究将产生新的仪器、新的成像探针和新的标记方法，所有这些都旨在对活细胞环境中蛋白质和RNA的行为进行新的化学洞察。这个高度跨学科的项目将为研究生和本科生提供研究机会。WEI小组的外展工作计划为K-12和本科生提供接触科学的机会。液-液相分离(LLP)的形成及其在生物功能中的作用已被广泛研究，有许多令人兴奋的新观察和发现。然而，仍然有很多东西需要了解，特别是关于生物大分子之间基本相互作用的生物物理和生化原理，特别是蛋白质和RNA以及它们形成凝聚体的能力。为了研究活细胞中关键的细胞内相分离凝聚物，WEI实验室正在开发新的受激拉曼光谱成像技术，这种技术是非侵入性的、定量的，并提供高空间和时间分辨率。具体地说，该研究计划的目标是：(1)开发功能性振动探针和标记方法，以量化体外和活细胞询问中已识别的LLP系统的组成和检测凝聚内相互作用；(2)建立新的仪器模块，以实现活细胞化学图谱的灵敏度提高20倍；和(3)用先进的数据分析进行深入的活细胞调查，以拼凑自然和复杂细胞环境中未知的化学特征和相互作用原理。该奖项由数学和物理科学局(MPS)化学部的CMI、CLP和CSDM-B计划以及生物科学局(BIO)分子和细胞生物科学部(MCB)的分子生物物理学计划共同资助。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，认为值得支持。