RoL: EAGER: DESYN-C3: Programmable control of metabolism in synthetic cells using intrinsically disordered proteins

几乎所有的蛋白质都具有特定的三维结构，以保持其在细胞中的特定活性。有一类蛋白质则不然。它们被称为内在无序蛋白（IDP）。它们在细胞中的作用似乎是自发地与其他蛋白质结合并共同激活它们。这是一个重要的功能，包括在设计一个合成细胞。该项目将探索IDP如何与其他蛋白质以及DNA和RNA相互作用。特别感兴趣的是这些相互作用如何影响基因表达和细胞代谢的调节。功能失调的相互作用与神经退行性疾病有关，如ALS（Lou Gehrig病）。该项目的结果可能会导致加强治疗和改善这些疾病患者的预后。该项目还将支持本科生和研究生的培训和教育。 代表性不足的群体的成员将通过科学参与计划参加。该项目将开发IDP和核酸之间的相互作用，作为基因表达的控制机制。该方法将扩展到蛋白质合成的调节。为了实现这一点，IDP将被修饰以结合和隔离无膜细胞器中的mRNA。IDP-RNA复合物将被设计为对环境线索，包括蛋白质浓度，温度和蛋白酶活性作出反应。它们还必须紧密结合靶RNA，并根据需要释放捕获的RNA用于表达。这些概念的测试将在无细胞蛋白质合成系统中进行。转录网络将通过表达工程化核酸结合蛋白的基因来增强。将创建途径来调节简单报告蛋白的表达，并构建电路来调节CRISPR干扰系统。因此，该项目将为合成细胞建立一个全球控制机制，使其功能适应可用的环境资源和线索。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。

Almost all proteins have a specific three-dimensional structure that maintains its specific activity in the cell. One class of proteins do not. They are referred to as intrinsically disordered proteins (IDPs). Their role in the cell appears to be to spontaneously associate with other proteins and activate them collectively. This is an important function to include in the design of a synthetic cell. This project will explore how IDPs interact with other proteins and with DNA and RNA. Of particular interest is how those interactions affect the regulation of gene expression and cell metabolism. Dysfunctional interactions are linked to neurodegenerative disorders, such as ALS (Lou Gehrig's disease). The results of this project may lead to enhanced treatments and improved outcomes for patients suffering from these diseases. The project will also support training and education of undergraduate and graduate students. Members of underrepresented groups will take part via science involvement programs. This project will develop interactions between IDPs and nucleic acids as a control mechanism for gene expression. The approach will be extended to the regulation of protein synthesis. To accomplish that, IDPs will be modified to bind and sequester mRNA in membrane-free organelles. The IDP-RNA complexes will be designed to be responsive to environmental cues, including protein concentration, temperature and protease activity. They must also bind the target RNA tightly, and release captured RNA for expression on demand. Testing of these concepts will take place in a cell-free protein synthesis system. Transcriptional networks will be augmented with genes that express engineered nucleic acid-binding proteins. Pathways will be created to regulate expression of a simple reporter protein and to build circuits to regulate CRISPR interference systems. Thus, this project will establish a global control mechanism for synthetic cells to adapt their functioning to available environmental resources and cues.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.