Amplification-free RNA Detection with CRISPR-Cas13

Overview

Journal Commun Biol

Specialty Biology

Date 2021 Apr 20

PMID 33875803

Citations 69

Authors

Hajime Shinoda

Yuya Taguchi

Ryoya Nakagawa

Asami Makino

Sae Okazaki

Masahiro Nakano

Yukiko Muramoto

Chiharu Takahashi

Ikuko Takahashi

Jun Ando

Takeshi Noda

Osamu Nureki

Hiroshi Nishimasu

Rikiya Watanabe

Affiliations

Soon will be listed here.

Abstract

CRISPR-based nucleic-acid detection is an emerging technology for molecular diagnostics. However, these methods generally require several hours and could cause amplification errors, due to the pre-amplification of target nucleic acids to enhance the detection sensitivity. Here, we developed a platform that allows "CRISPR-based amplification-free digital RNA detection (SATORI)", by combining CRISPR-Cas13-based RNA detection and microchamber-array technologies. SATORI detected single-stranded RNA targets with maximal sensitivity of ~10 fM in <5 min, with high specificity. Furthermore, the simultaneous use of multiple different guide RNAs enhanced the sensitivity, thereby enabling the detection of the SARS-CoV-2 N-gene RNA at ~5 fM levels. Therefore, we hope SATORI will serve as a powerful class of accurate and rapid diagnostics.

Citing Articles

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