文章基本信息

标题：CRISPR-Cas3-based diagnostics for SARS-CoV-2 and influenza virus
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作者：Kazuto Yoshimi ; Kohei Takeshita ; Seiya Yamayoshi 等
期刊名称：iScience
印刷版ISSN：2589-0042
出版年度：2022
卷号：25
期号：2
页码：1-14
DOI：10.1016/j.isci.2022.103830
语种：English
出版社：Elsevier
摘要：SummaryCRISPR-based diagnostics (CRISPR-dx), including the Cas12-based DETECTR and Cas13-based SHERLOCK Class 2 CRISPRs, have been used to detect the presence of DNA or RNA from pathogens, such as the 2009 pandemic influenza virus A (IAV) and the 2019 novel coronavirus SARS-CoV-2. Here, we describe the collateral single-stranded DNA cleavage with Class 1 type I CRISPR-Cas3 and highlight its potential for development as a Cas3-mediated rapid (within 40 min), low-cost, instrument-free detection method for SARS-CoV-2. This assay, which we call Cas3-Operated Nucleic Acid detectioN (CONAN), not only detects SARS-CoV-2 in clinical samples, but also offers specific detection of single-base-pair mutations in IAV variants. This tool allows rapid and accurate point-of-care testing for patients with suspected SARS-CoV-2 or drug-resistant IAV infections in hospitals.Graphical abstractDisplay OmittedHighlights•Type I CRISPR-Cas3 showstrans-cleavage activity on nearby nonspecific ssDNA•We developed CONAN as a method to easily and accurately detect viral RNA•CONAN provides rapid, low-cost, and instrument-free detection of SARS-CoV-2 and IAV•CONAN also allows the specific detection of single-base-pair mutationsDiagnostics; Virology