RoL: EAGER: DESYN-C3: Synthetic Biogenesis of Eukaryotic Cells

细胞核是细胞储存信息的地方。内质网（ER）是细胞制造蛋白质的地方。线粒体是细胞产生能量的地方。设计细胞来模仿人类细胞可能至少需要复制这些核心功能。本项目将尝试做到这一点。该项目将包括将人工染色体包装在一个由膜包围的隔间中，代表细胞核。细胞核将被一层折叠的膜结构包围，这种膜结构类似于内质网。内质网内部是制造蛋白质的机器。一个独立的膜室，与其他两个分离，将容纳产生能量的酶。所有这些将被包含在另一个膜室和一起，代表一个粗合成细胞。制造单独的隔室将是一项挑战，因为膜必须允许一些分子选择性地通过，同时阻止其他分子通过，而且有些膜需要具有复杂的形状。如果这个项目成功了，它将为后续的努力提供一个平台，使用这些类型的合成细胞来制造疫苗或杀死肿瘤，这只是列举了几种可能性。该项目将包括一个由当地高中生组成的团队，通过国际基因工程机器（iGEM）竞赛参与研究的各个阶段。该项目将开发设计细胞样结构所需的技术，使复杂的合成系统能够模仿生物细胞的结构和功能。有两项主要任务。首先，微流控技术将被用于在一个离散的水体积内组装一个确定的染色体群。一个核膜将在染色体周围形成，并与一个折叠的er样膜相互连接，该膜由促进曲率的蛋白质制成。从细胞中提取的生物成分将被整合到膜中。核膜将在细胞核内形成。然后将演示合成核内mRNA的转录。其次，线粒体样结构将被开发作为mRNA转录的ATP来源。基于囊泡的方法将支持通过线粒体-细胞核ATP运输对mRNA转录的外部控制。这个项目将有助于理解合成和连接其他复杂程度越来越高的细胞样细胞器的要求。这将为探索诸如细胞膜如何以及为什么细胞衰老等悬而未决的问题创造新的机会。合成细胞最终可用于DNA加工和基因调控的闭环控制，具有包括癌症免疫治疗在内的实际生物医学应用的巨大潜力。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。

The nucleus is where a cell stores information. The endoplasmic reticulum (ER) is where cells manufacture proteins. Mitochondria are where cells generate energy. Designing cells to mimic human cells will likely involve replicating at least those core functions. This project will attempt to do just that. The project will involve artificial chromosomes being packaged into a compartment surrounded by a membrane, representing the nucleus. The nucleus will be surrounded by a folded membrane structure that mimics the ER. Inside the ER will be machinery to produce proteins. A separate membrane compartment, unattached to the other two, will house enzymes for generating energy. All of these will be contained in another membrane compartment and together, represent a crude synthetic cell. Creating the individual compartments will be challenging because the membranes have to allow some molecules to pass through them selectively while preventing others, and because some membranes will need to have intricate shapes. If the project is successful, it will provide a platform for subsequent efforts to use these types of synthetic cells to manufacture vaccines or to kill tumors, just to name a couple of possibilities. The project will involve a team of local high school students in all phases of the research through the International Genetically Engineered Machine (iGEM) competition.This project will develop the techniques needed to engineer cell-like structures, enabling the development of complex synthetic systems mimicking both structure and function of biological cells. There are two major tasks. First, microfluidic techniques will be employed to assemble a defined population of chromosomes within a discrete aqueous volume. A nuclear envelope will be formed around the chromosomes and interconnected with a folded ER-like membrane engineered using curvature-promoting proteins. Biological components extracted from cells will be integrated into the membrane. A nuclear lamina will be formed within the nucleus. Transcription of mRNA within the synthetic nuclei will then be demonstrated. Second, mitochondria-like structures will be developed to serve as ATP sources for mRNA transcription. A vesicle-based approach will support external control over mRNA transcription through mitochondria-nucleus ATP transport. This project will contribute to understanding the requirements for synthesizing and linking other cell-like organelles with increasing levels of complexity. That will create new opportunities for exploring open questions such as membrane how and why cells age. Synthetic cells could ultimately be exploited for closed loop control over DNA processing and gene regulation, with significant potential for practical biomedical applications including cancer immunotherapy.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.