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

DREAM Sentinels：可复用且可编程的无细胞 ADAR 介导的 RNA 传感平台 (cfRADAR)，可快速、可扩展地响应突发病毒威胁

• 批准号: 2319913
• 负责人: Weihua Guan
• 金额: $ 70万
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2319913&HistoricalAwards=false
• 关键词: DREAM; Sentinels; Multiplexable; programmable; cell

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.

该项目旨在满足关键的公共卫生需求：病毒RNA的快速，精确检测，这是许多病毒的遗传蓝图，包括负责COVID-19的SARS-COV-2病毒。该方法是开发一种尖端工具，称为“作用于RNA上的无细胞可重编程腺苷脱氨酶”（CFRADAR）。该工具是一种高度适应性的病毒侦探，旨在快速修饰以识别新病毒或突变病毒。除了其主要功能外，Cfradar预计将产生更广泛的社会影响。首先，它可以通过提供有关病毒流行和传播的实时信息，实现早期检测，迅速对暴发的迅速反应以及最终降低传播风险的实时信息，从而显着改善医疗保健结果。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，该项目将从事外展和教育活动，为各个级别的学生和老师提供动手的经验和互动学习。因此，Cfradar的发展将显着受益于本地和全球社区，通过促进健康，繁荣和福利来为国家利益提供服务。该项目的主要目标是Cfradar的发展（无细胞可重编程的腺苷脱氨酸酶，作用于RNA上），RNA检测技术利用了腺苷酶的腺苷酶酶（ADNASPES）（ADNASES PARNAS od arna od arna od arna sonfar rna on rna on rna on rna on rna on rna on rna on rna od arna s ro. rna s ro. rna od rna on rna s ro. rna s ro. rna s ro.该项目将解决六个主要目标：1）开发单个复合物和2）多路复用CFRADAR分析，3）创建基于CFRADAR的逻辑电路，用于组测试，4）制定实地可部署的可部署冻干的Cfradar系统，5）设计现场可部署的可移植便携式分析仪，以及6）在基金 - f-File-Free-File-Free-Free-Free-Free-Free-Free-Free-Free-Free-Free-Free-Free-Free-Free-Free-Free-Free Free设置中验证CFRADAR。 Cfradar平台提供了诸如高灵敏度，重编程性，模块化和现场可部署性等优点。 Cfradar凭借其对多个传感器分子编辑的单目标转录物具有高灵敏度对低丰度RNA靶标的敏感性。它的模块化和重编程性允许使用布尔逻辑门，同时提供了多个RNA靶标的检测。对冻干的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，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，可重编程和可扩展的传感平台来彻底改变护理点诊断，小组测试和生物监视。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子优点和更广泛影响的审查标准来评估的。

项目成果

High Fidelity Machine-Learning-Assisted False Positive Discrimination in Loop-Mediated Isothermal Amplification Using Nanopore-Based Sizing and Counting

• DOI: 10.1021/acsnano.3c12053
• 发表时间: 2024-02-23
• 期刊: ACS NANO
• 影响因子: 17.1
• 作者: Dong，Ming;Kshirsagar，Aneesh;Guan，Weihua
• 通讯作者: Guan，Weihua