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Dynamic biosensing enables amplifier-collateral-cleavage enhancement for pathogen diagnostic.

Huiyou Chen1, Zhixi Zeng2, Yangdao Wei3

  • 1School of Marine Sciences, School of Life and Health Sciences, School of Marine Biology and Fisheries, School of Information and Communication Engineering, State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, Haikou, 570228, China; Department of Clinical Hematology, College of Pharmacy and Laboratory Medicine Science, Department of Clinical Laboratory Medicine, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, 400038, China.

Biosensors & Bioelectronics
|November 3, 2025
PubMed
Summary

CRISPR-CasΦ offers a novel biosensing approach by blocking collateral cleavage, enabling exponential signal amplification. This new method, Amplifier-collateral-cleavage enhancement (ACE), shows high sensitivity and specificity in clinical urine sample analysis.

Keywords:
CRISPR-CasΦDiagnosticDynamic biosensingUrinary pathogen

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • CRISPR Technology

Background:

  • CRISPR-Cas systems and DNA nanotechnology are vital in biomedicine.
  • Current methods face limitations in specialized experimental settings.

Purpose of the Study:

  • To explore the unique characteristics of CRISPR-CasΦ for advanced biosensing.
  • To develop a novel signal amplification strategy for enhanced detection.

Main Methods:

  • Investigated CRISPR-CasΦ's collateral-cleavage blockade mechanism.
  • Designed dynamic DNA-based signal amplifiers.
  • Developed the Amplifier-collateral-cleavage enhancement (ACE) method.

Main Results:

  • CRISPR-CasΦ exhibits collateral-cleavage blockade due to inability to recognize "TTN" sequences.
  • Developed two novel stem-loop amplifiers for CasΦ dynamic sensing.
  • ACE method achieved exponential signal amplification with 98.8% sensitivity and 90% specificity in clinical samples.

Conclusions:

  • CRISPR-CasΦ dynamic sensing presents a breakthrough for biosensing applications.
  • The ACE method offers a powerful tool for sensitive and specific detection.
  • This work bridges dynamic DNA technology and CRISPR systems for future innovations.