CRISPR/Cas12a-nanozyme visual biosensor for detection of microRNA-21

Area of Science:

• Biomedical Engineering
• Molecular Biology
• Biotechnology

Background:

• Accurate diagnosis of aggressive tumors is vital for effective treatment.
• MicroRNAs (miRNAs) are key biomarkers in cancer development and spread.
• Sensitive detection methods for specific miRNAs are needed for early diagnosis.

Purpose of the Study:

• To develop a novel CRISPR/Cas12a-nanozyme visual biosensor.
• To achieve direct and sensitive detection of tumor-associated miRNA-21.
• To explore its potential for point-of-care nucleic acid diagnostics.

Main Methods:

• Utilized a CRISPR/Cas12a system combined with a nanozyme (Ag/NiFe layered double hydroxide).
• Employed magnetic beads and single-stranded DNA (ssDNA) linkers for nanozyme immobilization.
• Incorporated strand displacement amplification (SDA) for signal amplification.
• Leveraged Cas12a activity to trigger a visible colorimetric reaction upon miRNA-21 detection.

Main Results:

• The biosensor demonstrated sensitive detection of miRNA-21 with a limit of 420.0 fM.
• Achieved direct visual readout, simplifying the detection process.
• The system successfully detected tumor-associated miRNA-21.

Conclusions:

• The developed CRISPR/Cas12a-nanozyme visual biosensor is a promising tool for sensitive miRNA detection.
• This approach offers a direct visual readout, suitable for point-of-care applications.
• The biosensor shows significant potential for early cancer diagnosis and monitoring.