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Related Experiment Video

Updated: Jan 27, 2026

DNA Virus Detection System Based on RPA-CRISPR/Cas12a-SPM and Deep Learning
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DNA Flap-Controlled CRISPR/Cas12a Trans-Cleavage Enables Mix-and-Read FEN1 Activity Detection.

Shengyong Ding1,2, Yuhang Li1, Fengshuang Wang2

  • 1School of Public Health, Shandong First Medical University and Shandong Academy of Medical Sciences, Jinan 250117, China.

Analytical Chemistry
|January 26, 2026
PubMed
Summary
This summary is machine-generated.

A new DNA flap-controlled CRISPR/Cas12a (FCT-CRISPR) method simplifies detecting flap endonuclease 1 (FEN1) activity. This innovation offers sensitive, accurate, mix-and-read detection without complex steps, aiding disease diagnosis.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Biosensing

Background:

  • CRISPR/Cas12a offers high specificity and signal amplification for biosensing.
  • Current methods for detecting flap endonuclease 1 (FEN1) activity using CRISPR/Cas12a require complex preligation or replication steps, increasing operational complexity and false signal risks.

Purpose of the Study:

  • To develop a simplified, sensitive, and accurate method for detecting FEN1 activity.
  • To establish a mix-and-read platform for FEN1 activity monitoring using CRISPR/Cas12a.

Main Methods:

  • A novel DNA flap-controlled CRISPR/Cas12a trans-cleavage (FCT-CRISPR) strategy was designed.
  • A flap-structured dumbbell DNA probe was utilized, where the flap acts as a split activator and the scaffold provides steric hindrance.
  • FEN1 cleavage liberates the activator, triggering CRISPR/Cas12a trans-cleavage for signal amplification.

Main Results:

  • The FCT-CRISPR strategy achieved sensitive and accurate detection of FEN1 activity.
  • The method demonstrated a low detection limit of 0.2 mU and high specificity against nontarget enzymes.
  • The platform successfully detected FEN1 activity in cancer cell lysates, showing clinical potential.

Conclusions:

  • FCT-CRISPR provides a sensitive, accurate, and mix-and-read platform for FEN1 activity detection.
  • This approach eliminates the need for exogenous DNA ligation or replication steps.
  • The FCT-CRISPR strategy shows promise for early diagnosis of FEN1-related diseases.