Search for the minimal requirements for sustainable and tunable cell cycles in synthetic cells

地球上几乎每个生物都有一个内在的生物钟，称为昼夜节律，驱动着许多重要身体功能的模式，比如动物的睡眠-觉醒周期。同样，所有细胞都依赖于另一个时钟，称为细胞周期，以精确地确定细胞生长和分裂的时间。虽然昼夜节律和细胞周期都已被研究了几十年，但这些生物钟本身就是与其他细胞网络相互作用的复杂网络，以协调各种下游事件，并经常与从失眠到癌症等疾病有关。这些系统的复杂性使得隔离核心机器以了解其最小化需求变得非常困难。该项目旨在通过重组和研究合成细胞中的最小细胞周期时钟系统，在理解细胞振荡的方向上取得进一步的重大进展。从这项研究中获得的知识通过出版物、会议、课程和实践演示向更广泛的社区传播。主要的更广泛的影响活动包括让代表性不足的学生及早接触前沿研究和留住这些学生在STEM领域的学习。大学水平的学生被介绍到在这个项目中开发的最先进的技术。该项目还制定和实施了外联方案，通过与密歇根大学国家历史博物馆的合作，向更广泛的公众传播科学发现。生物振荡器，包括细胞周期，对生物体在胚胎中建立形态发生模式和维持成人基本生理的能力有重大影响。因此，了解这些生物钟的设计和功能是至关重要的。这样的理解提供了开发的基本知识和创造合成振荡器的指导。尽管它很重要，但构建多功能和弹性振荡器的最小化设计原则仍然难以捉摸。这项研究结合了几种策略，以确定一个基本的失衡过程的基本规则，有丝分裂细胞周期。具体地说，该项目使用微乳液滴建立了一个独特的合成细胞系统，以剖析体外细胞周期网络的电路-功能关系，并使用所获得的知识自下而上地创建最小的细胞周期网络。合成的细胞系统由尽可能小的蛋白质、mRNAs和作为能量来源的三磷酸腺苷组成。一个成功的项目将提供对网络拓扑的最低要求的重要见解，该网络拓扑负责所需的细胞周期性能和对环境扰动的响应。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。

Almost every living being on Earth has an internal biological clock, known as circadian rhythms, to drive patterns of many important bodily functions, such as sleep-wake cycles in animals. Likewise, all cells rely on another clock, called cell cycles, for precise timing of cell growth and cell division. Although both circadian rhythms and cell cycles have been studied for decades, these biological clocks are themselves complicated networks interacting with other cellular networks to orchestrate various downstream events and are frequently connected to diseases from insomnia to cancer. The complexity of these systems makes it very difficult to isolate the core machinery to understand their minimized requirements. This project aims to make further major advances in the direction of understanding cellular oscillations by reconstituting and investigating minimal cell-cycle clock systems in synthetic cells. The knowledge gained from this research is disseminated to the broader community through publications, conferences, courses, and hands-on demonstrations. The major broader impact activities involve the early exposure of underrepresented students to frontier research and retention of these students in STEM fields. College level students are introduced to the state-of-the-art technology developed in this project. The project also develops and implements outreach programs to disseminate the scientific findings to the broader public through the collaborations with the University of Michigan Museum of National History. Biological oscillators, including the cell cycle, have a significant impact on the ability of organisms to establish morphogenetic patterns in embryos and maintain basic physiology in adults. It is thus crucial to understand the design and function of these biological clocks. Such understanding provides fundamental knowledge of development and guidance to create synthetic oscillators. Despite its importance, the minimized design principles for constructing versatile and resilient oscillators remains elusive. This research combines several strategies to ascertain the essential rules underlying a fundamental out-of-equilibrium process, the mitotic cell cycles. Specifically, the project builds a unique synthetic cell system, using microemulsion droplets, to dissect the circuit-function relation of an in vitro cell-cycle network and uses the knowledge gained to create a minimized cell-cycle network from the bottom up. The synthetic cellular system is composed of the smallest possible sets of proteins, mRNAs, and ATP as the energy source. A successful project will provide significant insights into the minimal requirements of the network topology that is responsible for the desired cell-cycle performance and response to environmental perturbations.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.