DREAM Sentinels: Multiplexable and programmable cell-free ADAR-mediated RNA sensing platform (cfRADAR) for quick and scalable response to emergent viral threats

该项目旨在解决一项关键的公共卫生需求：快速、精确地检测病毒RNA，即许多病毒的遗传蓝图，包括导致COVID-19的SARS-CoV-2病毒。该方法是开发一种称为“作用于RNA的无细胞可编程腺苷脱氨酶”（cfRADAR）的尖端工具。此工具的功能是作为一个高度适应性的病毒检测器，设计用于快速修改以识别新的或变异的病毒。除了其主要功能之外，预计cfRADAR还将产生更广泛的社会影响。首先，它可以提供有关病毒流行和传播的实时信息，从而显著改善医疗保健结果，实现更早的发现、对疫情的快速反应，并最终降低传播风险。这种可靠、快速的诊断工具可以增强我们预防、控制和治疗传染病的能力。其次，该项目将开展外联和教育活动，为不同层次的学生和教师提供实践经验和互动学习。因此，cfRADAR的发展将显著造福当地和全球社区，通过促进健康、繁荣和福利服务于国家利益。该项目的主要目标是开发cfRADAR（作用于RNA的无细胞可编程腺苷脱氨酶），这是一种利用作用于RNA的腺苷脱氨酶（ADAR）特性的RNA检测技术。该项目将解决六个主要目标：1)开发单路和2)多路cfRADAR分析，3)创建基于cfRADAR的逻辑电路用于组测试，4)制定可现场部署的冻干cfRADAR系统，5)设计可现场部署的便携式分析仪，6)在无基础设施的环境中验证cfRADAR。cfRADAR平台具有高灵敏度、可重新编程性、模块化和现场可部署性等优点。cfRADAR可将单靶点转录物编辑为多传感器分子，对低丰度RNA靶点具有较高的敏感性。它的模块化和可重新编程性允许使用布尔逻辑门，同时提供多个RNA目标的检测。冻干无细胞试剂和便携式分析仪的探索增强了平台的现场部署能力，使其成为及时应对病毒威胁的有效工具。该项目可以通过引入敏感、可重新编程和可扩展的传感平台，彻底改变即时诊断、小组测试和生物监测。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。

This project seeks to address a critical public health need: the rapid, precise detection of viral RNA, the genetic blueprint of many viruses, including the SARS-CoV-2 virus responsible for COVID-19. The approach is to develop a cutting-edge tool known as "cell-free reprogrammable adenosine deaminases acting on RNA" (cfRADAR). This tool functions as a highly adaptable virus detective, designed to be quickly modified to recognize new or mutating viruses. Beyond its primary function, cfRADAR is projected to have broader societal impacts. Firstly, it can significantly improve healthcare outcomes by providing real-time information on viral prevalence and spread, enabling earlier detection, prompt responses to outbreaks, and ultimately reducing the risk of transmission. This reliable, rapid diagnostic tool can enhance our capabilities to prevent, control, and treat infectious diseases. Secondly, the project will engage in outreach and educational activities, offering hands-on experiences and interactive learning for students and teachers at various levels. Therefore, cfRADAR's development stands to significantly benefit both local and global communities, serving the national interest by advancing health, prosperity, and welfare.This project's primary objective is the development of cfRADAR (cell-free reprogrammable adenosine deaminases acting on RNA), an RNA detection technology leveraging the properties of adenosine deaminases acting on RNA (ADAR). The project will tackle six main goals: 1) developing singleplex and 2) multiplex cfRADAR assays, 3) creating cfRADAR-based logic circuits for group testing, 4) formulating field deployable lyophilized cfRADAR systems, 5) designing a field deployable portable analyzer, and 6) validating cfRADAR in an infrastructure-free setting. The cfRADAR platform offers advantages such as high sensitivity, reprogrammability, modularity, and field deployability. With its single-target transcript to multiple sensor molecule editing, cfRADAR possesses high sensitivity towards low-abundance RNA targets. Its modularity and reprogrammability allow for the use of Boolean logical gates, offering the detection of multiple RNA targets simultaneously. The exploration of lyophilized cell-free reagents and portable analyzers enhances the platform's field deployment, making it an effective tool for timely response against viral threats. The project could revolutionize point-of-care diagnosis, group tests, and biosurveillance by introducing a sensitive, reprogrammable, and scalable sensing platform.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.