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Split G-quadruplex-programmed label-free CRISPR-Cas12a sensing system.

Mengting Xu1, Yuedong Zhu1, Yan Zhang1

  • 1School of Pharmacy, Nantong University, Nantong, Jiangsu 226001, China. yuyanyan@ntu.edu.cn.

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|May 31, 2023
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Summary

A novel Cas12a biosensing platform using split G-quadruplex motifs was developed. This label-free system accurately detects and profiles exosomal proteins for cancer diagnostics.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Cas12a nucleases offer programmable DNA targeting capabilities.
  • G-quadruplex (G4) structures can be integrated into nucleic acid-based biosensors.
  • Detecting exosomal surface proteins is crucial for cancer biomarker discovery.

Purpose of the Study:

  • To establish and optimize a split G-quadruplex (G4)-programmed Cas12a platform for biosensing.
  • To develop a label-free detection method for target molecules.
  • To demonstrate the platform's utility in profiling exosomal surface proteins from cancer samples.

Main Methods:

  • Engineering a split G4 motif to serve as a Cas12a substrate.
  • Utilizing Cas12a activity for signal generation in a label-free manner.
  • Applying the platform to detect and analyze exosomal surface proteins.

Main Results:

  • Successful establishment, validation, and optimization of the split G4-Cas12a platform.
  • Demonstrated a concentration-dependent, label-free response to target molecules.
  • Successfully detected and profiled exosomal surface proteins from both cultured cancer cells and clinical samples.

Conclusions:

  • The developed split G4-Cas12a platform is a sensitive and specific tool for molecular detection.
  • This label-free biosensing approach holds promise for exosomal protein analysis in cancer diagnostics.
  • The platform's versatility supports potential applications in biomarker discovery and clinical settings.