Photo-switchable CRISPR electrochemical system enables de-interferential biosensing signal output

Area of Science:

• Biomedical Engineering
• Molecular Diagnostics
• CRISPR Technology

Background:

• CRISPR-based electrochemical biosensors often suffer from signal crosstalk, leading to false positives.
• A controllable switching function is needed to improve the reliability and accuracy of these biosensors.

Purpose of the Study:

• To develop a photo-switchable CRISPR/Cas12a electrochemical (PSCE) system for precise gene mutation detection.
• To mitigate signal crosstalk and enhance the accuracy of electrochemical biosensing.

Main Methods:

• Incorporation of a photocleavable (PC) linker and a light-responsive strategy into the CRISPR/Cas12a system.
• Decoupling nucleic acid amplification (NAA) from signal transduction for a photo-switchable response.
• Utilizing light irradiation to activate Cas12a activity and separate signal readout from sample pretreatment.

Main Results:

• The PSCE system demonstrated de-interference electrochemical signals, achieving high sensitivity and accuracy for non-small cell lung cancer (NSCLC) gene mutation detection.
• The system suppressed intrinsic amplification-derived signal crosstalk, resulting in an ultralow limit of detection (LoD).
• Achieved 98.1% sensitivity, 92.7% specificity, and 98.9% overall accuracy with 67 clinical samples.

Conclusions:

• The developed PSCE system offers a reliable and accurate method for bioanalysis by addressing signal crosstalk.
• The system shows significant potential for clinical diagnosis of mutation-associated diseases, including applications in wearable electronics and machine learning analysis.