New Technologies for detecting extracellular fluxes

检测细胞外通量的新技术

基本信息

批准号：
10456044
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/10456044
关键词：
Action Potentials Animals Astrocytes Bacteria Benchmarking Brain Cardiac Cell membrane Cells Coculture Techniques Communication Data 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 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：这种重点技术和发展赠款的目标是复制细胞内荧光离子和代谢物传感器可检测细胞外通量。三个正交接近直接靶向细胞的糖脂：纳米厚，糖涂在离子的地方在离子通道和膜转运蛋白期间，代谢物浓度差异很大活动。 AIM 1描述了将任何荧光蛋白传感器连接到细胞的方法代谢标记，并带有不自然的氮杂糖。 AIM 2是一种非遗传学方法标记所有哺乳动物细胞（包括人类细胞），以及小分子和蛋白质 AIM 3是一种实现细胞类型特异性标记的化学遗传方法带有小分子和蛋白质荧光传感器。目标的完成将转换为这些使用三个主要类别的荧光传感器（小分子，蛋白质和fret）进入使离子和代谢物可视化的技术原代细胞，组织的表面，并可能在活动物中。