New Technologies for detecting extracellular fluxes

检测细胞外通量的新技术

基本信息

批准号：
10649530
负责人：
WILLIAM R KOBERTZ
金额：
$ 36.42万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10649530
关键词：
Action Potentials Animals Astrocytes Bacteria Benchmarking Brain Cardiac Cell membrane Cells Coculture Techniques Communication Data Development Disease Engineering Fluorescence Fluorescence Resonance Energy Transfer Glutamates Glycocalyx Goals Grant Health Homeostasis Human Individual Ion Channel Ions Kinetics Label Mammalian Cell Membrane Potentials Membrane Proteins Membrane Transport Proteins Metabolic Microglia Muscle Muscle Contraction Muscle function Neurons Pathway interactions Periodicity Physiological Positioning Attribute Proteins Quality Control Reagent Research Personnel Rest Series Signal Transduction Surface Techniques Technology Thick Time Tissues Viral Visualization Wheat Germ Agglutinins cell fixing cell type chemical genetics extracellular genetic approach heart function human disease nanometer neuronal excitability new technology non-genetic polyhistidine prevent prototype sensor small molecule spatiotemporal sugar technology development technology research and development uptake

项目摘要

Abstract: The goal of this Focused Technology and Development grant is to repurpose intracellular fluorescent ion and metabolite sensors to detect extracellular fluxes. The three orthogonal approaches directly target the cell’s glycocalyx: the nanometer thick, sugar‐coating where ion and metabolite concentrations vary greatly during ion channel and membrane transporter activity. Aim 1 describes an approach to attach any fluorescent protein sensor to cells metabolically-labeled with an unnatural azidosugar. Aim 2 is a non-genetic approach that labels all mammalian cells—including human cells—with both small molecule and protein fluorescent sensors. Aim 3 is a chemical genetic approach to achieve cell-type-specific labeling with small molecule and protein fluorescent sensors. Completion of the aims will convert these working prototypes that utilize three major classes of fluorescent sensors (small molecule, protein, and FRET) into technologies that enable the visualization of ions and metabolites at the surface of a primary cells, tissues, and potentially in living animals.

抽象的：这项重点技术和开发拨款的目标是重新利用细胞内荧光离子和代谢物传感器检测细胞外通量。方法直接针对细胞的糖萼：纳米厚的糖衣，其中离子离子通道和膜转运过程中代谢物浓度变化很大目标 1 描述了一种将任何荧光蛋白传感器附着到细胞上的方法。 Aim 2 是一种非遗传方法，用非天然叠氮糖进行代谢标记。用小分子和蛋白质标记所有哺乳动物细胞（包括人类细胞） Aim 3 是一种实现细胞类型特异性标记的化学遗传方法。与小分子和蛋白质荧光传感器的完成将转换这些。利用三大类荧光传感器（小分子、蛋白质和 FRET）转化为能够实现离子和代谢物可视化的技术原代细胞、组织的表面，也可能存在于活体动物的表面。