Characterizing dynamic properties and responses of an artificial mitotic cell

有丝分裂是细胞分裂和增殖的重要过程。一个细胞分成两个，两个细胞分成四个，以此类推。这个过程是由一组基因和蛋白质精细控制的。这些被称为有丝分裂振荡器。有丝分裂的缺陷被认为会增加患癌症的几率。研究细胞内的振荡器是很困难的，因为细胞内还发生着许多其他复杂的细胞过程。这个项目将把一个合成的有丝分裂振荡器制造成微小的液滴来模拟真实的细胞。没有细胞生长和细胞分裂的复杂影响，这些微滴可以定量操作和表征有丝分裂振荡器。通过这项研究获得的知识将通过出版物、会议和讲习班向更广泛的科学界传播。与本地专业教育工作者和博物馆合作，透过拟议的外展活动，例如示范和实验室开放日，向青少年和公众传播跨学科的技术。这些活动将使我们能够吸引对科学感兴趣的6-9年级少数民族学生，让他们接触到研究的激情，并为他们从事STEM职业做好准备。本项目的目的是定量分析最小有丝分裂振荡器的随机动力学及其对各种环境信号的响应。为了构成最小的细胞质有丝分裂振荡器，爪蟾卵细胞质提取物将被封装在模拟单细胞行为的微流体油包水乳剂中。此外，一个包含细胞核的更复杂的细胞将建立在这个最小的振荡器之上。这将概括体外复杂的现象，如核组装、染色质凝聚和蛋白质定位。几个问题将被检查在这个系统中，包括作用的电池尺寸，温度，和能量消耗在调制振荡器的性质。为了实现这些目标，计算建模、基于微流体的方法和延时荧光成像将被整合。这项工作将自下而上地构建一个循环细胞，范围从最简单的不含细胞核的形式到驱动各种核活动的复杂形式。这项工作的成功将为寻找药物开发和再生医学预防和治疗有丝分裂振荡相关癌症疾病的新靶点提供有价值的指导，因此对癌症生物学，细胞和发育生物学的更广泛社区具有重要意义。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。

Mitosis is the important process by which cells divide and proliferate. A cell divides into two, and two into four, and so on. This process is finely controlled by a set of genes and proteins. These are known as the mitotic oscillator. Defects in mitosis are thought to increase the chance of developing cancer. It is difficult to study the oscillator inside the cell, where many other complicated cellular processes are also occurring. This project will build a synthetic mitotic oscillator into tiny droplets to mimic real cells. Without the complicated effects from cell growth and cell divisions, these microdroplets enable quantitative manipulation and characterization of the mitotic oscillator. The knowledge gained through this research will be disseminated to the broader scientific community through publications, conferences, and workshops. The interdisciplinary technologies will be disseminated to youth and the public through proposed outreach activities such as demonstrations and lab open day in collaboration with local professional educators and the museum. These activities will allow us to engage underrepresented minority students in grades 6-9 interested in science, expose them to research excitement, and prepare them for STEM careers.The goal of this project is to quantitatively analyze the stochastic dynamics of a minimal mitotic oscillator and its responses to various environmental signals. To constitute a minimal cytoplasmic mitotic oscillator, Xenopus egg cytosolic extracts will be encapsulated in microfluidic water-in-oil emulsions that mimic single cell behaviors. Additionally, a more complicated cell that contains nuclei will be built on top of this minimal oscillator. This will recapitulate complex phenomena in vitro such as nuclear assembly, chromatin condensation, and protein localization. Several questions will be examined in this system, including the role of cell size, temperature, and energy depletion in modulating properties of the oscillator. To achieve these goals, computational modeling, droplet-based microfluidic methods, and time-lapse fluorescence imaging will be integrated. This work will build a cyclic cell bottom-up, ranging from the simplest form containing no nuclei to the complicated ones driving various nuclear activities. The success of this work will provide valuable guidance in search of new targets for drug development and regenerative medicine in preventing and treating mitotic oscillator-related cancerous diseases, and thus be relevant and of great importance to broader communities in cancer biology, and cell and developmental biology.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.