BBSRC-NSF/BIO - Deciphering the rules of nucleus architecture with synthetic cells and organelles

细胞系统的特征之一是生物分子含量的非均匀空间分布。虽然在所有的生命类中都可以发现内容物的区室化，但它在含有各种亚细胞器的真核生物中是一个特别显著的特征。细胞器是存在于不同化学空间中的细胞内的离散结构，并且专门执行定义的生化任务。占优势的真核细胞器是容纳基因组并确保转录和翻译的空间分离的细胞核。人们认为，真核生物在细胞核和其他细胞器中划分功能的能力是其行为复杂性的背后。然而，技术的局限性意味着我们对控制细胞中分区的基本设计原则缺乏基本的理解。为什么细胞要对内容进行分区？什么定义子隔间的大小和数量？核结构如何影响DNA转录和蛋白质合成？是什么决定了细胞核的大小和细胞大小之间的关系？近年来，自下而上的合成生物学（bottom-up synthetic biology）作为基础生物科学中一个强大的新研究学科出现：通过构建合成细胞，重新设计生物学，以破译生命规则。合成细胞是在形式、功能和行为上模仿生物细胞的结构。它们是通过将生物分子块以确定的组合方式聚集在一起来复制细胞生命的各个方面：通过构建新的生物学来理解生物学。近年来，这一研究领域取得了显著的发展，部分原因是我们在帝国理工和加州理工的实验室采用了先进的新技术来模拟、操纵和制造新的生物分子系统。在这个国际合作项目中，我们的英国和美国团队将同时开发新的微流体和合成生物学技术，以组装含有核样细胞器的合成细胞。结合新的数学模型，我们将使用我们的合成细胞来研究上述与细胞生物学有关的未解决的基本问题。通过解构和重建细胞核的关键细胞结构基序，我们的目标是解开真核生物结构的基本原则，更普遍的细胞区室化。我们将开发的建模和实验平台将是多功能的、可推广的、开放获取的和技术化的，使我们的技术和工具包能够被更广泛的学术界利用，以利用合成细胞应对各种生物学挑战。虽然这个项目将侧重于与基础生物学相关的问题，但我们将根据我们的见解开发工程规则手册，结合我们将开发的新技术，可以用于构建新一代的合成细胞装置，在临床和工业中用作智能治疗剂，生物反应器和生物修复。此外，该计划促进的独特跨大西洋合作也将成为美国和英国大型财团之间的桥梁，致力于从零开始构建合成细胞的巨大挑战。

One of the hallmarks of cellular systems is the non-uniform spatial distribution of biomolecular content. Although compartmentalisation of content can be found across all life classes, it is a particularly marked feature in eukaryotes which contain a diverse array of subcellular organelles. Organelles are discrete structures within cells that exist in distinct chemical space, and are specialised to perform defined biochemical tasks. The dominant eukaryotic organelle is the nucleus which houses the genome and ensures spatial segregation of transcription and translation. It is thought that the ability of eukaryotes to compartmentalise functions in the nucleus and other organelles is behind their behavioural sophistication. Technological limitations, however, have meant that our fundamental understanding of the underpinning design principles that govern compartmentalisation in cells is lacking.Why do cells compartmentalise content? What defines the size and number of sub-compartments? How does nuclear architecture affect DNA transcription and protein synthesis? What governs the relationship between the size of the nucleus and the size of a cell? There is a need for new experimental and modelling technologies to shed light on these questions.Recent years have seen the emergence of bottom-up synthetic biology as a powerful new research discipline in the fundamental biosciences: re-engineering biology to decipher the rules of life by building synthetic cells. Synthetic cells are structures that mimic biological cells in form, function and behaviour. They are made by bringing together biomolecular blocks in defined combination to replicate aspects of cellular life: understanding biology by building a new biology. This research area has seen remarkable growth in recent times, partly driven by pioneering new technologies in our labs at Imperial and Caltech to model, manipulate and make new biomolecular systems.In this collaborative and international project, our UK and US teams will work in parallel to develop new microfluidic and synthetic biology technologies to assemble synthetic cells that contain nucleus-like organelles. Together with integration with new mathematical models, we will use our synthetic cells to investigate the above unresolved fundamental questions relating to cell biology. By deconstructing and reconstructing the key cellular architectural motif of the nucleus, we aim to unravel the fundamental principles of eukaryotic architecture, and of cellular compartmentalisation more generally. The modelling and experimental platforms we will be developing will be versatile, generalisable, open access, and deskilled, allowing our enabling technologies and toolkits to be utilised by the wider academic community to tackle diverse biological challenges using synthetic cells.Although this project will focus on questions relating to fundamental biology, the engineering rule-book we will be developing as a result of our insights, combined with the novel technologies we will be developing can be deployed for the construction of a new generation of synthetic cell devices with uses in the clinic and industry as smart therapeutic agents, as bioreactors, and in bioremediation. Moreover, the unique trans-Atlantic collaboration facilitated by this scheme will also serve as a bridge between large-scale consortia in the USA and UK dedicated to the grand challenge of building a synthetic cell from scratch.