RoL: RAISE: DESYN-C3: Membrane-based cell-free systems for scalable production of plant natural products

制作出能够工作的东西意味着开发者能够从根本上理解它是如何工作的，并且能够添加一些特性去研究它们对性能的影响。例如，对一个正在工作的飞机模型，我们可以在机翼上加一条曲线，增加翼尖的缺口，以及一系列的修改，看看它们是如何影响飞机的性能的。同样的设计范式也适用于生物学。如果一个人可以从零开始制造一个可以工作的细胞，那么他就可以开始给它添加碎片，并在系统中设计功能。该研究项目旨在创造一种模拟植物细胞膜的合成细胞系统。一旦实现了一个功能系统，通常在植物细胞膜上发现的不同蛋白质将被添加到合成细胞系统中，以确定它们显示的功能。研究人员将主要关注产生阿尔茨海默病药物的蛋白质模型集。为了进一步提高研究项目的影响，研究人员还将开发有关合成细胞的新教材，培训研究生和本科生，并向公众进行宣传，以产生和进一步了解合成细胞的概念及其潜在用途。迄今为止，还不可能创造出自组装的、基于膜的无细胞系统。本研究项目的主要假设是，可以通过调节磷脂囊泡生成酶的时间和表达水平来实现成功。为了实现这一目标，研究人员将把基于crispr的遗传控制系统与磷脂生产途径结合起来。具体而言，他们将构建一个高效的植物跨膜蛋白的膜基因表达系统。拟合成的植物天然产物是治疗阿尔茨海默病的药物加兰他明。如果成功生产加兰他明，研究人员将把该系统扩展到各种其他跨膜蛋白质，模拟植物细胞环境。因此，该研究项目将扩大无细胞生物工程的能力，并将提供新的方法来制备具有复杂膜组成的合成细胞系统。这些系统将使科学家更好地了解膜生物物理和酶活性之间的关系。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。

Making something that works signifies that one understands how it works at a fundamental level and that one can add features to study their effects upon performance. For example, to a working airplane model one can add a curve to the wings, up-notched wingtips, and a whole host of modifications to see how they affect the performance of the plane. The same design paradigm holds true in biology. If one can make a cell from scratch that works, then one can start adding pieces to it and engineering functions into the system. This research project aims to create a synthetic cell system that mimics the membrane of a plant cell. Once a functioning system is achieved, different proteins that normally are found in plant cell membranes will be added to the synthetic cell system to determine what functions they display. The researchers will primarily focus on model set of proteins that produce an Alzheimer's drug. To further enhance the impact of the research project, the researchers will also develop new educational material about synthetic cells, train graduate and undergraduate students, and perform outreach to the general public to generate and further understanding about the idea of synthetic cells and their potential uses. To date, it has not been possible to create self-assembling, membrane-based cell-free systems. The primary hypothesis of this research project is that success can be achieved through regulated control over the timing and expression levels of enzymes for phospholipid vesicle generation. To achieve this goal, the researchers will integrate a CRISPR-based genetic control system with phospholipid production pathways. Specifically, they will construct a highly-efficient membrane-based gene expression system for plant transmembrane proteins. The intended plant natural product to be synthesized is the Alzheimer's drug, Galantamine. If successful in producing Galantamine, the researchers will expand the system to a variety of other transmembrane proteins, emulating plant cell environments. As a result, this research project will expand cell-free bioengineering capabilities and will provide new methods to prepare synthetic cell systems with complex membrane compositions. These systems will enable scientists to better understand the relationship between membrane biophysics and enzymatic activity.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.