RoL: RAISE: DESYN-C3: Engineering multi-compartmentalised synthetic minimal cells

该项目旨在通过建立对细胞器如何在自然细胞中产生和维持的新见解来创建用于构建功能性人工细胞的工具。乍一看，真核细胞器可以被描述为细胞内的小隔间，由称为脂质的分子组成的膜从细胞内容物的其余部分中划分出来。 细胞器对细胞的生命至关重要，反过来，它们的形成取决于脂质的自组装成膜和不同液相的共存。因此，脂质的自组装和液-液相共存是目前研究的重点。相比之下，这两个过程之间的相互作用在很大程度上是未知的。有了这个奖项，化学系的生命过程化学项目和化学、生物工程、环境和运输系统系的细胞和生物化学工程项目正在资助宾夕法尼亚州立大学的克莉丝汀博士和明尼苏达大学的凯特·阿达马拉博士研究两种主要类型的细胞内组织过程如何相互作用。这项跨学科的研究为人工细胞器的构建和人工细胞的长期发展奠定了基础。这项研究也为研究生、本科生和K-12教师的科学培训和专业发展提供了机会。本研究项目探索了细胞质模拟系统，其中脂质和相分离蛋白质通过无细胞转录/翻译酶促产生。体外脂质生物合成途径被开发用于在模型细胞质中原位脂质生成。这些脂质的自组装模型无膜细胞器的存在下，其特征在于通过光学和电子显微镜，以及通过渗透性测量。该项目旨在设计能够产生功能性合成脂质膜和诱导性应激颗粒的人工细胞。这些发现推进了对器官发生的物理机制的理解，以及局部脂质产生和介质区室化如何影响这些机制。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

This project aims to create tools for construction of functional artificial cells by building on new insight into how organelles are generated and maintained in natural cells. At a glance, eukaryotic organelles could be described as small intracellular compartments delineated from the rest of the cell content by a membrane made up of molecules called lipids. The organelles are critical for the life of the cell and in turn their formation depends on the self-assembly of the lipids into membranes and on the coexistence of different liquid phases. Consequently the lipid self-assembly and the coexistence of liquid-liquid phases are the object of on-going investigations. In contrast, the interactions between these two processes are largely unexplored. With this award, the Chemistry of Life Processes Program in the Division of Chemistry and the Cellular and Biochemical Engineering Program in the Division of Chemical, Bioengineering, Environmental and Transport Systems are funding Dr. Christine Keating from Penn State University and Dr. Kate Adamala from the University of Minnesota to investigate how the two main types of intracellular organizational processes interact with each other. This interdisciplinary research generates a stepping stone on the path to the construction of artificial organelles and in the long term of artificial cells. The research also represents an opportunity for the scientific training and professional development of graduate students, undergraduates, and K-12 teachers. This research project explores cytoplasm-mimicking systems in which lipids and phase-separating proteins are enzymatically produced by cell-free transcription/translation. In vitro lipid biosynthesis pathways are developed for in situ lipid generation in model cytoplasm. The self-assembly of these lipids in the presence of model membraneless organelles is characterized by optical and electron microscopies as well as by permeability measurements. This project seeks to engineer artificial cells capable of producing functional synthetic lipid membranes and inducible stress granules. The findings advance the understanding of physical mechanisms for organellogenesis, and how local lipid production and media compartmentalization impact these mechanisms.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.