EAGER: Engineered, Smart, Nucleic Acid-Binding, Intrinsically Disordered Proteins to Enable Ubiquitous Detection of Viral Pathogens and Diagnosis

在过去十年中，出现了几次致命的病毒性疾病流行病，其中最引人注目的是COVID-19。防止这些疾病传播的关键一线防线是在受感染人群和其他环境中快速检测病毒的能力。该项目解决了基础研究的迫切需要，使有前途的新型高性能技术能够检测特定病毒。用于检测病毒核糖核酸（rna）的低成本、可广泛部署的新检测策略可大大增强对流行病和大流行传播的了解和缓解。这个跨学科的研究项目为研究生和本科生，包括代表性不足的群体的成员，提供了发展研究技能和跨学科工作的机会。本研究探索了新的方法来显著提高关键步骤和生物分析方法的性能，这些方法是基于聚合酶链反应扩增的耗时测试的替代方法。在该项目的第一个目标中，研究团队利用基因工程，生物启发，内在无序蛋白（IDPs）的相变行为来集中和增强快速，有效的病毒RNA分离。第二个目标是探索IDP作为伴侣有效催化RNA杂交和链置换反应的潜在用途。这些反应是病毒核酸检测方法的核心，如CRISPR-Cas9触发的链位移扩增、支点开关和分子逻辑电路。这个项目的目标是方便、大规模地部署低成本的即时病毒检测和诊断。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。

The past decade has seen the emergence of several epidemics of deadly viral diseases, most notably, COVID-19. A key front-line defense against the spread of these diseases is the ability to rapidly detect viruses in infected humans, and in other contexts. This project addresses the urgent need for fundamental research that enables promising new, high performance technologies for detection of specific viruses. Low cost, widely deployable new testing strategies for detection of viral ribonucleic acids (RNAs) can greatly enhance understanding and mitigation of the spread of epidemics and pandemics. This interdisciplinary research project provides opportunities to graduate and undergraduate students, including members of underrepresented groups, to develop research skills and to work across disciplines.This research explores novel approaches to significantly enhance key steps and the performance of bioanalytical methods that are alternatives to the time-consuming testing based on polymerase chain reaction amplification. In the first aim of this project, the research team capitalizes on the phase change behavior of genetically engineered, bioinspired, intrinsically disordered proteins (IDPs) to concentrate and enhance rapid, efficient viral RNA isolation. The second aim explores the potential use of IDP as chaperones to effectively catalyze RNA hybridization and strand displacement reactions. These reactions are central to viral nucleic acid detection methodologies such as CRISPR-Cas9 triggered strand displacement amplification, toehold switches, and molecular logic circuits. The goal of this project is to enable facile, massive deployment of low-cost, point-of-care virus detection and diagnosis.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.