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iScience
. 2022 Jan 30;103830.
doi: 10.1016/j.isci.2022.103830. Online ahead of print.
CRISPR-Cas3-based diagnostics for SARS-CoV-2 and influenza virus
Kazuto Yoshimi 1 2 , Kohei Takeshita 3 , Seiya Yamayoshi 4 , Satomi Shibumura 5 , Yuko Yamauchi 1 , Masaki Yamamoto 3 , Hiroshi Yotsuyanagi 6 , Yoshihiro Kawaoka 4 7 , Tomoji Mashimo 1 2
Affiliations
- PMID: 35128347
- PMCID: PMC8801231
- DOI: 10.1016/j.isci.2022.103830
Abstract
CRISPR-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.