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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Related Experiment Video

Updated: May 1, 2026

DNA Virus Detection System Based on RPA-CRISPR/Cas12a-SPM and Deep Learning
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High-efficiency detection of APE1 using a defective PAM-driven CRISPR-Cas12a self-catalytic biosensor.

Yang Song1, Juan Long2, Huaibi Wang1

  • 1Department of Oncology, Chongqing Hospital of Traditional Chinese Medicine, Chongqing, 400016, China.

Biosensors & Bioelectronics
|March 29, 2025
PubMed
Summary

A new CRISPR-Cas12a strategy, DEP-Cas-APE, uses defective PAMs for sensitive detection of apurinic/apyrimidinic endonuclease 1 (APE1) activity. This method enables rapid, isothermal, one-step molecular diagnostics and point-of-care testing for cancer biomarkers.

Keywords:
APE1 detectionCRISPR-Cas12aDefective PAMPoint-of-care testing (POCT)Self-catalytic signal amplification

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Diagnostics

Background:

  • CRISPR-Cas systems offer sensitive molecular diagnostics but face limitations in catalytic efficiency and require complex preamplification.
  • Detecting apurinic/apyrimidinic endonuclease 1 (APE1) activity is crucial for molecular diagnostics and understanding DNA repair pathways.

Purpose of the Study:

  • To develop a novel, highly sensitive, and specific CRISPR-Cas12a-based strategy for detecting apurinic/apyrimidinic endonuclease 1 (APE1) activity.
  • To create an efficient, isothermal, one-step detection method that overcomes the limitations of conventional CRISPR-Cas diagnostics.

Main Methods:

  • A defective PAM-mediated CRISPR-Cas12a self-catalytic signal amplification strategy (DEP-Cas-APE) was developed using modified DNA probes.
  • The strategy integrates Cas12a trans-cleavage with a self-catalytic circuit for signal amplification under isothermal conditions.
  • A point-of-care testing (POCT) platform was created by combining DEP-Cas-APE with a gold nanoparticle-based colorimetric assay.

Main Results:

  • The DEP-Cas-APE strategy achieved a low detection limit of 7.66 × 10-8 U μL-1 for APE1 activity within 30 minutes.
  • The method demonstrated high sensitivity and specificity in detecting APE1 in complex biological samples, including human serum.
  • The integrated POCT platform enabled portable, equipment-free detection and successfully distinguished between cancerous and normal serum samples.

Conclusions:

  • DEP-Cas-APE provides a robust and versatile platform for rapid, sensitive, and specific molecular diagnostics using CRISPR-Cas12a technology.
  • This strategy offers significant potential for advancing biosensing technologies and enabling new opportunities in clinical research and diagnostics.
  • The developed POCT platform facilitates accessible, equipment-free disease biomarker detection.