CRISPR-Cas3-based diagnostics for SARS-CoV-2 and influenza virus

Kazuto Yoshimi; Kohei Takeshita; Seiya Yamayoshi; Satomi Shibumura; Yuko Yamauchi; Masaki Yamamoto; Hiroshi Yotsuyanagi; Yoshihiro Kawaoka; Tomoji Mashimo

iScience (Feb 2022)

CRISPR-Cas3-based diagnostics for SARS-CoV-2 and influenza virus

Kazuto Yoshimi,
Kohei Takeshita,
Seiya Yamayoshi,
Satomi Shibumura,
Yuko Yamauchi,
Masaki Yamamoto,
Hiroshi Yotsuyanagi,
Yoshihiro Kawaoka,
Tomoji Mashimo

Affiliations

Kazuto Yoshimi: Division of Animal Genetics, Laboratory Animal Research Center, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan; Division of Genome Engineering, Center for Experimental Medicine and Systems Biology, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan
Kohei Takeshita: Advanced Photon Technology Division, RIKEN SPring-8 Center, Hyogo 679-5148, Japan
Seiya Yamayoshi: Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan
Satomi Shibumura: C4U Corporation, Osaka 565-0871, Japan
Yuko Yamauchi: Division of Animal Genetics, Laboratory Animal Research Center, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan
Masaki Yamamoto: Advanced Photon Technology Division, RIKEN SPring-8 Center, Hyogo 679-5148, Japan
Hiroshi Yotsuyanagi: Division of Infectious Diseases and Applied Immunology, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan
Yoshihiro Kawaoka: Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan; Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53711, USA
Tomoji Mashimo: Division of Animal Genetics, Laboratory Animal Research Center, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan; Division of Genome Engineering, Center for Experimental Medicine and Systems Biology, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan; Corresponding author

Journal volume & issue: Vol. 25, no. 2
p. 103830

Abstract

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Summary: 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.

Published in iScience

ISSN: 2589-0042 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science
Website: http://www.cell.com/iscience/home

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