Japan_IPAP - Top-down meets bottom-up: Designer membrane-less organelles from condensation of synthetic RNA nanostructure

Japan_IPAP - 自上而下与自下而上相遇：通过合成 RNA 纳米结构的浓缩设计无膜细胞器

• 批准号: BB/X012557/1
• 负责人: Lorenzo Di Michele
• 金额: $ 19.27万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FX012557%2F1
• 关键词: Japan_IPAP; Top; down; meets; bottom

Eukaryotic cells are characterised by the presence of internal compartments, or organelles, that by separating or bringing together specific biomolecular components can optimise processes critical for the cells to survive. Many organelles, such as the nucleus, the mitochondria and the Golgi apparatus are enclosed by lipid membranes, that have long been considered as the only means through which cells can establish internal compartments. However, in recent years, a new class of organelles has been discovered, which are not bound by a membrane. These membrane-less organelles emerge thanks to the ability of certain proteins and RNA molecules to "separate" from the rest of the cytoplasm, forming droplets due to the same physical process that causes oil droplets to separate from water. Like membrane-bound organelles, these biomolecular droplets or "condensates" play important biological roles, regulating for example, the assembly of enzymes and natural degradation of RNA.Synthetic biology aims to re-program cells to enhance their functionalities, making them useful as therapeutic or sensing agents, and as "cell factories" for the optimised production of pharmaceutical compounds and biomaterials. Cell reprogramming is most often conducted through genetic engineering: re-writing the cell's DNA to change its behaviour.Because of the importance that compartmentalisation plays in regulating cellular functionalities, the possibility of controlling the formation and composition of organelles would be very valuable in synthetic biology, complementing the tools of genetic engineering.Thanks to an interdisciplinary research team from the UK and Japan, in this project, we will develop strategies to engineer the formation of non-native membrane-less organelles in cells. The new organelles will assemble from of RNA molecules that fold to form nanostructures with prescribed shape and mutual interactions. We will be able to program the RNA-organelles to capture other biomolecules, such as messenger RNA, enzymes, and other proteins, so to replicate the ability of natural organelles to spatially organise and regulate the cell's biochemical pathways. We will study the properties of the RNA organelles at first in simplified cell models: synthetic cells that mimic the properties of live cells. Later, we will move to induce the formation of the RNA organelles in live cells, specifically E. coli bacteria. Note that, as all prokaryotes, E. coli do not possess natural internal compartmentalisation, so we would be creating completely new structures within the cell!Our findings will give synthetic biologists a new tool to fine tune the behaviour of live cells, which we argue could be particularly useful for optimising bioprocessing and biosynthesis.

真核细胞的特征是存在内部区室或细胞器，通过分离或聚集特定的生物分子成分可以优化细胞生存的关键过程。许多细胞器，如细胞核、线粒体和高尔基体，都被脂质膜包围，长期以来，脂质膜一直被认为是细胞建立内部区室的唯一手段。然而，近年来，人们发现了一类新的细胞器，它们不受膜的束缚。这些无膜细胞器的出现得益于某些蛋白质和 RNA 分子与细胞质其余部分“分离”的能力，由于导致油滴与水分离的相同物理过程而形成液滴。与膜结合细胞器一样，这些生物分子液滴或“凝聚物”发挥着重要的生物学作用，例如调节酶的组装和 RNA 的自然降解。合成生物学旨在重新编程细胞以增强其功能，使它们可用作治疗药物或传感剂，以及作为优化生产药物化合物和生物材料的“细胞工厂”。细胞重编程通常通过基因工程进行：重写细胞的 DNA 以改变其行为。由于区室化在调节细胞功能中的重要性，控制细胞器的形成和组成的可能性在合成生物学中将非常有价值，补充基因工程工具。感谢来自英国和日本的跨学科研究团队，在这个项目中，我们将开发策略来工程化细胞中非天然无膜细胞器的形成。新的细胞器将由 RNA 分子组装而成，这些分子折叠形成具有指定形状和相互作用的纳米结构。我们将能够对RNA细胞器进行编程，以捕获其他生物分子，例如信使RNA、酶和其他蛋白质，从而复制天然细胞器在空间上组织和调节细胞生化途径的能力。我们将首先在简化的细胞模型中研究 RNA 细胞器的特性：模仿活细胞特性的合成细胞。稍后，我们将着手诱导活细胞（特别是大肠杆菌）中 RNA 细胞器的形成。请注意，与所有原核生物一样，大肠杆菌不具有自然的内部区室化，因此我们将在细胞内创建全新的结构！我们的发现将为合成生物学家提供一个新工具来微调活细胞的行为，我们认为对于优化生物加工和生物合成特别有用。