RAPID: Point-of-Need Detection of COVID-19 using CRISPR-Enabled Cell-Free Synthetic Biology

RAPID：使用支持 CRISPR 的无细胞合成生物学对 COVID-19 进行定点检测

基本信息

批准号：
2028651
负责人：
Julius Lucks
金额：
$ 20万
依托单位：
Northwestern University
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-01 至 2022-04-30
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=2028651&HistoricalAwards=false
关键词：
RAPID Point Need Detection COVID

项目摘要

The COVID-19 pandemic highlights a limitation of laboratory-based testing for the SARS-CoV-2 coronavirus in that it does not scale with a sudden and dramatic increase in volume. Laboratory tests, often based on quantitative polymerase chain reaction (qPCR), require equipment, time, expertise and infrastructure, resulting in severe logistical challenges and ultimately resulting in inadequate testing. There is a critical need for diagnostics that can be rapidly deployed at the point-of-need (PON) to enable global surveillance of infectious diseases. Towards addressing this need, this project incorporates a new CRISPR-based approach called CRISPR Isothermal Amplification (CIA) that enables one-pot diagnostic amplification and detection of the SARC-CoV-2 virus at ambient temperatures. This scheme builds off innovations in cell-free synthetic biology and biosensing that enable rapid, low-cost, and PON detection of chemical contaminants and viral pathogens. The platform technology under development could serve several purposes from a quick-screen technology to inform triage and isolation strategies, to a clinical test used on the front lines. There is also the potential for the technology to eventually serve as an at-home test to help inform which subpopulations and sectors of the economy could safely return to productivity towards the end of the pandemic. In addition, the test has the flexibility to be rapidly reprogrammed to detect new emerging pathogens. This work is pursued in close collaboration with a commercial partner, Stemloop, Inc., in order to quickly transition the technology for manufacturing, deployment, and distribution.This project employs a new CRISPR Isothermal Amplification (CIA) approach to enable a one-pot amplification and detection of the SARS-CoV-2 genome in an assay for the virus. CIA uses programmed Cas 13 systems to recognize SARS-CoV-2 RNA, trigger nucleic acid amplification and create a detectable output simultaneously in the same reaction where amplification occurs. Computational models of each process are developed alongside the experimental systems and used to guide optimizations to achieve attomolar sensitivity. The method is unique in that no DNA primers are required. The amplification and detection processes are designed to take less than one hour and cost less than one dollar per test to manufacture.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.

COVID-19大流行强调了SARS-COV-2冠状病毒的基于实验室测试的局限性，因为它不会随着体积的突然而显着增加而扩展。实验室测试通常基于定量聚合酶链反应（QPCR）需要设备，时间，专业知识和基础设施，从而导致严重的后勤挑战，并最终导致测试不足。对于诊断的迫切需要，可以在需要的地方快速部署（PON），以实现全球对传染病的监视。为了满足这一需求，该项目结合了一种新的基于CRISPR的方法，称为CRISPR等热放大（CIA），该方法在环境温度下实现了一锅诊断放大和检测SARC-COV-2病毒。该方案建立了无细胞合成生物学和生物传感的创新，从而使化学污染物和病毒病原体的快速，低成本和PON检测能够进行快速，低成本和PON检测。正在开发的平台技术可以实现多种目的，从快速屏幕技术为分类和隔离策略提供信息，到前线使用的临床测试。该技术最终也有可能作为家庭测试，以帮助告知哪些亚种群和经济领域可以安全地恢复大流行的生产力。此外，该测试具有快速重编程以检测新的新兴病原体的灵活性。与商业合作伙伴STEMLOOP，Inc。密切合作，以快速过渡到制造，部署和分销的技术。该项目采用一种新的CRISPR等温扩增（CIA）方法来启用对ViRUS的SARS-COV-2基因组进行单盘扩增和检测。 CIA使用编程的CAS 13系统识别SARS-COV-2 RNA，触发核酸扩增并在发生扩增的同一反应中同时创建可检测的输出。每个过程的计算模型都与实验系统一起开发，并用于指导优化以实现Attomall敏感性。该方法是独一无二的，因为不需要DNA引物。放大和检测过程旨在花费不到一个小时，每次测试的成本不到一美元。该奖项反映了NSF的法定任务，并使用基金会的知识分子优点和更广泛的影响审查标准，被认为值得通过评估来获得支持。