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Dynamic DNA assembly-assisted CRISPR/Cas12a system for lung cancer-associated miRNA analysis
Anzhi Sheng1, Bingjie Zeng2, Haizhen Jin1
1Department of Central Laboratory, Shanghai Chest Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200030, PR China.
Materials Today. Bio
|April 11, 2025
Summary
This study introduces a novel CRISPR/Cas12a-based method for ultrasensitive detection of lung cancer biomarkers (miRNAs). The innovative approach significantly enhances early cancer diagnostics and prognostic evaluation through precise molecular detection.
Area of Science:
- Molecular Diagnostics
- Biomarker Discovery
- CRISPR Technology
Background:
- Serum microRNAs (miRNAs) are crucial biomarkers for early cancer detection and prognosis.
- Ultrasensitive quantification of miRNAs is essential for advancing molecular diagnostics.
- Existing analytical platforms require improvement for enhanced sensitivity.
Purpose of the Study:
- To develop a novel strategy for ultrasensitive detection of lung cancer-associated miRNAs.
- To leverage the CRISPR/Cas12a system for amplified signal generation.
- To establish a robust platform for early cancer diagnostics.
Main Methods:
- Utilized a functional hairpin DNA template (H) for target miRNA recognition.
- Initiated an isothermal exponential amplification reaction (iEXPAR) upon miRNA binding.
- Activated the CRISPR/Cas12a system for signal amplification using generated amplicons.
Main Results:
- Achieved ultrasensitive detection of lung cancer-associated miRNAs.
- Demonstrated a linear detection range from 20 fM to 2 nM.
- Obtained a low detection limit of 26 fM.
Conclusions:
- The proposed H-sequence and CRISPR/Cas12a strategy enables highly sensitive miRNA detection.
- This platform shows significant potential for early lung cancer diagnosis and prognostic evaluation.
- The DNA sequence programmability allows for adaptation to detect various nucleic acid targets.
