A membrane encapsulation system for cellular reconstitution

用于细胞重建的膜封装系统

• 批准号: 8965379
• 负责人: DANIEL A FLETCHER
• 金额: $ 29.66万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8965379
• 关键词: Acoustics; Address; Area; Biochemical Reaction; Biological; Biotechnology; Caliber; Cell Size; Cell physiology; Cells; Cellular Structures; Cellular biology; Complex; Complex Mixtures; Coupling; DNA; Detergents; Device or Instrument Development; Drug Targeting; Encapsulated; Environment; Epidermal Growth Factor Receptor; Extracellular Space; Future; G-Protein-Coupled Receptors; Genes; Genetic; Human; Image; In Vitro; Integral Membrane Protein; Intracellular Space; Kinetics; Length; Life; Ligands; Ligation; Lipids; Liquid substance; Measurement; Mediating; Membrane; Membrane Proteins; Methods; Microfluidic Microchips; Microfluidics; Modeling; Molecular; Molecular Conformation; Pilot Projects; Process; Production; Property; Proteins; Quality Control; Reaction; Receptor Signaling; Research Personnel; Sampling; Second Messenger Systems; Side; Signal Transduction; Signaling Molecule; Solutions; Stimulus; System; Techniques; Technology; Tertiary Protein Structure; Time; Variant; Vesicle; Work; base; design; extracellular; flexibility; in vivo; inhibitor/antagonist; instrument; internal control; protein complex; protein function; public health relevance; receptor; reconstitution; repaired; research study; screening; second messenger

 DESCRIPTION: Advances in imaging, genetics, and sequencing have made it possible to identify, track, and manipulate molecules involved in many different cellular processes in living cells. But are our models of these processes complete and correct at the molecular level? And can we understand mechanistically why these processes fail and what might be required to repair them? This is especially a challenge for processes that involve membrane proteins, such as those that mediate interactions with extracellular signals. Even though membrane proteins comprise ~30% of human genes and are important targets for drugs, the techniques available for characterizing and manipulating them are severely limited. What is needed to answer basic and applied questions about membrane protein function is a technology for re-building cell-like systems from the bottom up that recapitulates the spatial organization, confinement, and dynamic properties of living cells. This project will develop a new instrument capable of creating membrane encapsulations with oriented membrane proteins, offering a new experimental platform for answering questions about how information is transmitted across membranes. By encapsulating complex mixtures of proteins, DNA, and other biomolecules inside bilayer membrane vesicles with high throughput and high yield, the proposed instrument will address a fundamental obstacle in our understanding of how membrane proteins, beginning with EGFR and GPCRs, organize and respond to stimuli.

 产品说明：成像、遗传学和测序的进步使得识别、跟踪和操纵参与活细胞中许多不同细胞过程的分子成为可能。但是，我们对这些过程的模型在分子水平上是完整和正确的吗？我们能否从机制上理解这些过程失败的原因，以及修复它们可能需要什么？这对于涉及膜蛋白的过程尤其是一个挑战，例如介导与细胞外信号相互作用的那些。尽管膜蛋白包含约30%的人类基因，并且是药物的重要靶点，但可用于表征和操纵它们的技术受到严重限制。要回答有关膜蛋白功能的基本和应用问题，需要一种自下而上重建细胞样系统的技术，该技术概括了活细胞的空间组织、限制和动态特性。该项目将开发一种新的仪器，能够用定向膜蛋白创建膜结构，为回答有关信息如何跨膜传输的问题提供一个新的实验平台。通过以高通量和高产率将蛋白质、DNA和其他生物分子的复杂混合物封装在双层膜囊泡内，所提出的仪器将解决我们理解膜蛋白（从EGFR和GPCR开始）如何组织和响应刺激的根本障碍。