A membrane encapsulation system for cellular reconstitution

 描述：成像、遗传学和测序的进步使识别、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？我们能从机械上理解这些过程失败的原因以及修复它们可能需要什么吗？对于涉及膜蛋白的过程来说，这尤其是一个挑战，例如那些调节与细胞外信号相互作用的过程。尽管膜蛋白约占人类基因的30%，是药物的重要靶点，但可用于表征和操纵它们的技术严重有限。回答有关膜蛋白功能的基本和应用问题所需要的是一种自下而上重建类细胞系统的技术，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，能够用定向的膜蛋白创建膜包膜，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包裹在双层膜小泡中，所提出的仪器将解决我们理解膜蛋白如何组织和响应刺激的根本障碍，从EGFR和GPCRs开始。

 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.