RoL: EAGER: DESYN-C3: Emergent mechanics of synthetic cells

人体内的每个细胞都包含一个由肌动蛋白（一种蛋白质）细丝网络组成的结构框架。这个框架就像一个骨架。它是细胞抵抗变形、改变形状和移动能力的基础。我们对这种肌动蛋白细胞骨架的理解是基于对整个细胞行为的研究。然而，我们并不完全了解这些丝状网络是如何组装的。这个项目将重组肌动蛋白网络，并观察它们在压力下如何重组自己。肌动蛋白也将被合成，并在许多环境条件下观察到其自组织成结构。该项目将提供深入了解肌动蛋白细胞骨架网络结构的机制，以生产强大的合成细胞。该项目还将为从高中到研究生院的学生提供研究机会。将特别强调为K-12一级的女孩开展的外联活动。关于细胞感知和响应力以及获得机械刚度的能力如何来自细胞骨架构建模块，我们知之甚少。理解这一点将使合成细胞中的机械功能设计成为可能。 重建平台可以阐明肌动蛋白结构如何影响细胞的机械特性。本项目将使用两个模型系统。一种是合成细胞，它包裹着重组的细胞骨架成分。第二种是脂质双层囊泡，其封装产生肌动蛋白的无细胞蛋白质合成（CFPS）系统。这些将通过能够施加机械压缩的新型微流体装置进行处理，并且将评估细胞/囊泡和肌动蛋白网络的响应。将量化具有不同肌动蛋白网络的合成细胞的机械性质。基于这些结果，将进行并评估具有可重构肌动蛋白网络的合成细胞的工程化。这种可重构的内部细胞骨架的展示将为工程刺激响应合成细胞的变化铺平道路。这项工作可能会揭示新的规则，通过突出一个强大的合成细胞的机械要求的合成细胞的设计。如果成功的话，这些结果将导致合成细胞的设计，对外部机械扰动的反应更鲁棒。这是目前自下而上构建合成电池的一个主要障碍。该奖项反映了NSF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。

Each cell in the human body contains a structural framework that is composed of a network of actin (a protein) filaments. This framework acts like a skeleton. It is the basis of the cell's ability to resist deformation, to change shape, and to move. Our understanding of this actin cytoskeleton is based on studies of the behavior of whole cells. However, we do not fully understand how these filamentous networks assemble. This project will reconstitute actin networks and observe how they reorganize themselves under stress. Actin will also be synthesized and its self-organization into structures will be observed under many environmental conditions. The project will provide insight into the mechanics of actin cytoskeleton network construction for production of robust synthetic cells. The project will also provide research opportunities for students from high school through graduate school. Of specific emphasis will be outreach activities for girls at the K-12 level. Very little is known about how the ability of cells to sense and respond to force and acquire mechanical stiffness arises from cytoskeletal building blocks. Understanding this would enable the design of mechanical functionality in synthetic cells. A reconstitution platform could elucidate how actin architectures affect the mechanical properties of a cell. Two model systems will be used in this project. One involves synthetic cells that encapsulate reconstituted cytoskeletal components. The second is a lipid bilayer vesicle that encapsulates a cell-free protein synthesis (CFPS) system that produces actin. These will be processed through a novel microfluidic device capable of exerting mechanical compression, and the responses of the cell/vesicle and of the actin network will be evaluated. The mechanical properties of synthetic cells with different actin network will be quantified. Based on those results, the engineering of a synthetic cell with reconfigurable actin networks will be undertaken and evaluated. This demonstration of a reconfigurable internal cytoskeleton will pave the way for engineering stimulus-responsive changes to synthetic cells. This work may reveal new rules about synthetic cell design by highlighting the mechanical requirement of a robust synthetic cell. If successful, the results will lead to the design of synthetic cells with a more robust response to external mechanical perturbations. This is currently a major hurdle in the bottom-up construction of synthetic cells.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.