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Rapid and Accurate Detection of Novel Coronavirus SARS-CoV-2 Using CRISPR-Cas3

29 Pages Posted: 2 Jul 2020 Sneak Peek Status: Review Complete

See all articles by Kazuto Yoshimi

Kazuto Yoshimi

University of Tokyo - Division of Animal Genetics

Kohei Takeshita

RIKEN SPring-8 Center - Life Science Research Infrastructure Group

Seiya Yamayoshi

University of Tokyo - Division of Virology

Satomi Shibumura

C4U Corporation

Yuko Yamauchi

University of Tokyo - Division of Animal Genetics

Masaki Yamamoto

RIKEN SPring-8 Center - Life Science Research Infrastructure Group

Hiroshi Yotsuyanagi

University of Tokyo - Division of Virology

Yoshihiro Kawaoka

University of Wisconsin - Madison - Department of Pathobiological Sciences

Tomoji Mashimo

University of Tokyo - Division of Animal Genetics

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Abstract

Novel coronavirus SARS-CoV-2 outbreaks have rapidly spread to multiple countries, highlighting the urgent necessity for fast, sensitive, and specific diagnostic tools for virus surveillance. Here, the previously unknown collateral single-stranded DNA cleavage we observed with type I CRISPR-Cas3 highlights its potential for development as a Cas3-mediated rapid (within 40 min), low-cost, instrument-free detection method for SARS-CoV-2. This Cas3-based assay is comparable with Cas12- and real-time reverse-transcriptase PCR-based assays in its speed and sensitivity, but offers greater specificity for single-base-pair discrimination while negating the need for highly trained operators. These findings support the use of CRISPR diagnostics for point-of-care testing in patients with suspected SARS-CoV-2 infections.

Funding: This project was supported in part by JSPS KAKENHI Grant Number 18H03974 (T.M.) and 19K16025 (K.Y.). Partially support also came from the Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Supporting Innovative Drug Discovery and Life Science Research (BINDS)) from AMED under Grant Number JP20am0101070 (support numbers 1251 and 2463), from Research Program on Emerging and Reemerging Infectious Diseases from AMED (JP19fk0108113), and from the National Institutes of Allergy and Infectious Diseases funded Center for Research on Influenza Pathogenesis (CRIP; HHSN272201400008C).

Conflict of Interest: S.S. is employee of C4U. K.Y. and T.M. are co-founders and scientific advisors for C4U. The other authors declare no competing interests.

Suggested Citation

Yoshimi, Kazuto and Takeshita, Kohei and Yamayoshi, Seiya and Shibumura, Satomi and Yamauchi, Yuko and Yamamoto, Masaki and Yotsuyanagi, Hiroshi and Kawaoka, Yoshihiro and Mashimo, Tomoji, Rapid and Accurate Detection of Novel Coronavirus SARS-CoV-2 Using CRISPR-Cas3. Available at SSRN: https://ssrn.com/abstract=3640844
This is a paper under consideration at Cell Press and has not been peer-reviewed.

Kazuto Yoshimi

University of Tokyo - Division of Animal Genetics

Japan

Kohei Takeshita

RIKEN SPring-8 Center - Life Science Research Infrastructure Group ( email )

Seiya Yamayoshi

University of Tokyo - Division of Virology ( email )

Yayoi 1-1-1
Bunkyo-ku
Tokyo, Tokyo 113-8657
Japan

Satomi Shibumura

C4U Corporation ( email )

Yuko Yamauchi

University of Tokyo - Division of Animal Genetics ( email )

Japan

Masaki Yamamoto

RIKEN SPring-8 Center - Life Science Research Infrastructure Group ( email )

Hiroshi Yotsuyanagi

University of Tokyo - Division of Virology ( email )

Yayoi 1-1-1
Bunkyo-ku
Tokyo, Tokyo 113-8657
Japan

Yoshihiro Kawaoka

University of Wisconsin - Madison - Department of Pathobiological Sciences

716 Langdon Street
Madison, WI 53706-1481
United States

Tomoji Mashimo (Contact Author)

University of Tokyo - Division of Animal Genetics ( email )

Japan

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