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Updated: Oct 25, 2025

Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases
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Accelerated RNA detection using tandem CRISPR nucleases.

Tina Y Liu1,2, Gavin J Knott1,2,3, Dylan C J Smock1,2

  • 1Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.

Nature Chemical Biology
|August 6, 2021
PubMed
Summary
This summary is machine-generated.

This study introduces a new CRISPR-based method for rapid, direct RNA detection without amplification. The Fast Integrated Nuclease Detection In Tandem (FIND-IT) assay enables sensitive detection of RNA, including SARS-CoV-2, in just 20 minutes.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Diagnostics

Background:

  • Direct, amplification-free RNA detection is crucial for transforming molecular diagnostics and enabling on-site analysis.
  • Existing CRISPR-Cas systems for RNA detection face limitations in reaction time, affecting sensitivity and speed.

Purpose of the Study:

  • To develop a novel CRISPR-based assay for rapid and sensitive direct RNA detection.
  • To overcome the limitations of current CRISPR-Cas systems by enhancing detection speed and sensitivity.

Main Methods:

  • Utilized a tandem system of unrelated CRISPR nucleases (Cas13 and Csm6) for RNA sensing and signal generation.
  • Incorporated a chemically stabilized activator to create a one-step assay.
  • Developed the Fast Integrated Nuclease Detection In Tandem (FIND-IT) approach.

Main Results:

  • Achieved robust detection of approximately 30 molecules per µl of RNA within 20 minutes.
  • Demonstrated sensitive detection of SARS-CoV-2 RNA from respiratory swab samples with quantitative reverse transcriptase PCR (qRT-PCR)-derived cycle threshold (Ct) values up to 33.
  • Successfully employed a compact detector for the assay.

Conclusions:

  • The FIND-IT approach enables sensitive, direct RNA detection.
  • This method is suitable for point-of-care infection diagnosis and various other diagnostic or research applications.
  • The FIND-IT system offers a significant advancement in rapid molecular diagnostics.