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 大流行凸显了基于实验室的 SARS-CoV-2 冠状病毒检测的局限性，因为它不会随着数量的突然大幅增加而扩大。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，导致严峻的后勤挑战，并最终导致测试不充分。迫切需要能够在需求点 (PON) 快速部署的诊断技术，以实现对传染病的全球监测。为了满足这一需求，该项目采用了一种基于 CRISPR 的新方法，称为 CRISPR 等温扩增 (CIA)，该方法能够在环境温度下对 SARC-CoV-2 病毒进行一锅诊断扩增和检测。该计划建立在无细胞合成生物学和生物传感方面的创新基础之上，能够对化学污染物和病毒病原体进行快速、低成本的 PON 检测。正在开发的平台技术可用于多种用途，从用于告知分诊和隔离策略的快速筛选技术，到前线使用的临床测试。该技术还有可能最终作为家庭测试，帮助了解哪些亚人群和经济部门可以在大流行结束时安全地恢复生产力。此外，该测试具有快速重新编程的灵活性，以检测新出现的病原体。这项工作是与商业合作伙伴 Stemloop, Inc. 密切合作进行的，以便快速过渡制造、部署和分销技术。该项目采用了一种新的 CRISPR 等温扩增 (CIA) 方法来实现一锅法在病毒检测中扩增和检测 SARS-CoV-2 基因组。 CIA 使用编程的 Cas 13 系统来识别 SARS-CoV-2 RNA，触发核酸扩增，并在发生扩增的同一反应中同时产生可检测的输出。每个过程的计算模型都是与实验系统一起开发的，并用于指导优化以实现阿托摩尔灵敏度。该方法的独特之处在于不需要 DNA 引物。扩增和检测过程的设计时间不到一小时，每次测试的制造成本不到一美元。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力优点和更广泛的影响审查标准进行评估，被认为值得支持。