Collaborative Research: ProteoCell: The Fat-Free Cell

该项目旨在通过使用蛋白质创建多个隔室的“蛋白质细胞”来构建具有功能的、无脂的“活”细胞，该“蛋白质细胞”可以自我生成、容纳反应以产生产品、与其他蛋白质细胞相互作用，并总体而言，定义一种具有根本新颖特性的新型类细胞结构。细胞是定义生命系统的最基本单位，然而，尽管进行了多年的研究，但仍不清楚为什么大自然选择使用脂类作为其标志性的隔室形成者，以及隔室作用如何使复杂的生物分子作为一个自主和自我维持的单位发挥作用。这个项目通过使用工程学原理来设计新的蛋白质合成反应、新的隔室形成蛋白质和新的蛋白质催化剂来解决这些问题，并通过研究这些“设计的”蛋白质分子如何相互作用来创建集体细胞样单位。这项工作将解决围绕着脂类和蛋白质作为基本细胞‘构件’的作用的基本生命规则问题，以及细胞功能区隔的重要性。合成蛋白质细胞最终可能提供一种创新工具来解决与人类健康、可持续性和环境有关的社会问题。ProteoCell平台的开发还提供了独特的教育机会和机会，与公众进行双向对话，讨论合成细胞的含义，以及与创造新形式的细胞生命相关的可能性、好处和关注事项。从科学上讲，该项目将定义如何“自下而上”构建无脂蛋白质细胞，包括自组装物理边界、可遗传编程的细胞质和封装的多组分反应。蛋白质细胞将提供对多肽/蛋白质构建块的动态合成、它们的自组装以及创建最终能够自我生成和自我支持的功能性蛋白质细胞的设计规则之间的关系的见解。该项目方法包括膜形成多肽/蛋白质的模块化和迭代工程、生产和整合；用于二氧化碳转化的蛋白质机械；以及支持基因表达的机械。该项目的四个目标将创建可组成的蛋白质细胞并定义可能的性质范围；构建基本细胞器并定义从二氧化碳生产有机分子所需的性质；建立具有明确组织的多细胞蛋白质细胞‘社区’；以及探索美国公众对构建合成细胞的风险/好处的看法。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.