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

人体中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。这个框架的作用就像一个骨架。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。这个项目将重建肌动蛋白网络，并观察它们在压力下如何自我重组。肌动蛋白也将被合成，并将在许多环境条件下观察到其自组织成结构。该项目将提供对肌动蛋白细胞骨架网络构建机制的洞察，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。特别强调的是为女孩举办的K-12级外联活动。关于细胞感知和响应力以及获得机械硬度的能力是如何从细胞骨架构件中产生的，我们知之甚少。了解这一点将使合成细胞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性能。本项目将使用两个模型系统。其中一个涉及包裹重组细胞骨架成分的合成细胞。第二个是脂质双层囊泡，它包裹了一个产生肌动蛋白的无细胞蛋白质合成(CFPS)系统。这些将通过一种能够施加机械压缩的新型微流控装置进行处理，并将评估细胞/囊泡和肌动蛋白网络的响应。具有不同肌动蛋白网络的合成细胞的力学性能将被量化。在这些结果的基础上，将进行和评估具有可重构肌动蛋白网络的合成细胞的工程。这一可重新配置的内部细胞骨架的演示将为设计合成细胞对刺激反应的改变铺平道路。这项工作可能会通过强调坚固的合成电池的机械要求来揭示合成电池设计的新规则。如果成功，这些结果将导致合成电池的设计，使其对外部机械扰动的反应更强劲。这是目前合成细胞自下而上构建的一个主要障碍。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。

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.