Collaborative Research: ProteoCell: The Fat-Free Cell

该项目旨在通过使用蛋白质创造多室“ProteoCells”来构建功能性的、无脂的“活”细胞，这种细胞可以自我生成，进行反应以产生产品，与其他ProteoCells相互作用，总的来说，定义一种具有基本新颖特性的新型细胞样结构。细胞是定义生命系统的最基本单位，然而，尽管经过多年的研究，人们仍然不清楚为什么大自然选择使用脂质作为其标志性的区隔形成物，以及区隔化如何使生物分子的复杂“汤”作为一个自主和自我持续的单位发挥作用。该项目通过使用工程原理设计新的蛋白质合成反应、新的区室形成蛋白质和新的蛋白质催化剂，并通过研究这些“设计师”蛋白质分子如何相互作用以创造集体细胞样单位，来解决这些问题。这项工作将解决围绕脂质和蛋白质作为基本细胞“构建块”的作用的基本生命规则问题，以及细胞功能区隔化的重要性。合成蛋白质细胞可能最终为解决与人类健康、可持续发展和环境有关的社会问题提供一种创新工具。ProteoCell平台的开发也提供了独特的教育机会和机会，与公众就合成细胞的意义，以及与创造新形式的细胞生命相关的可能性，益处和关注进行双向对话。科学上，该项目将定义如何“自下而上”构建无脂蛋白细胞，包括自组装物理边界、基因可编程细胞质和封装的多组分反应。ProteoCells将提供多肽/蛋白质构建块的动态合成，它们的自组装和设计规则之间的关系，以创建最终可以自我生成和自我支持的功能ProteoCells。项目方法包括模块化和迭代工程，生产和膜形成多肽/蛋白质的整合；二氧化碳转化的蛋白质机制；以及支持基因表达的机制。该项目的四个目标是创建可组合的ProteoCells并定义可能的属性范围；构建基本的细胞器并确定从二氧化碳中产生有机分子所需的性质；构建具有明确组织的多细胞ProteoCell“社区”；并调查美国公众对构建合成细胞的风险/收益的看法。该奖项反映了美国国家科学基金会的法定使命，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 seeks to build functional, lipid-free, 'living' cells by using proteins to create multi-compartment 'ProteoCells' that can self-generate, house reactions to produce products, interact with other ProteoCells, and in total, define a new type of cell-like structure with fundamentally novel properties. Cells are the most basic unit defining living systems, yet despite years of study, it remains unclear why nature chose to use lipids as its hallmark compartment-formers, and how compartmentalization enables a complex 'soup' of biological molecules to function as an autonomous and self-sustainable unit. This project addresses these questions by using engineering principles to design new protein synthesis reactions, new compartment-forming proteins, and new protein catalysts, and by studying how these 'designer' protein molecules interact with one another to create collective cell-like units. This work will address fundamental Rules of Life questions surrounding the roles of lipids and proteins as essential cellular 'building blocks,' and the importance of compartmentalization in cellular function. Synthetic ProteoCells may ultimately provide an innovative tool to solve societal problems related to human health, sustainability, and the environment. Development of the ProteoCell platform also provides unique educational opportunities and opportunities to engage in a two-way dialog with the public regarding the meaning of a synthetic cell, and the possibilities, benefits, and concerns related to creating new forms of cellular life.Scientifically, this project will define how to 'bottom-up' construct lipid-free ProteoCells, including a self-assembling physical boundary, a genetically programmable cytoplasm, and encapsulated multicomponent reactions. ProteoCells will provide insights into the relationship between the dynamic synthesis of polypeptide/protein building blocks, their self-assembly, and the design rules to create functional ProteoCells that can ultimately self-generate and self-support. The project approach consists of modular and iterative engineering, production, and integration of membrane-forming polypeptides/proteins; protein machinery for CO2 transformation; and machinery to support gene expression. The four aims of the project will create composable ProteoCells and define the range of possible properties; construct rudimentary organelles and define the properties necessary for producing organic molecules from CO2; build multi-cellular ProteoCell 'communities' with defined organization; and probe the US public's perceptions of risks/benefits of constructing a synthetic cell.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.