CRISPR/Cas12a-based fluorescence aptasensor integrated with two-dimensional cobalt oxyhydroxide nanosheets for IFN-γ...

Area of Science:

• Biotechnology
• Molecular Biology
• Immunology

Background:

• Cytokine storm (CS) involves dangerous immune overreactions with elevated pro-inflammatory cytokines like interferon-gamma (IFN-γ).
• Sensitive detection of cytokines is crucial for understanding CS progression and diagnosing infectious diseases.

Purpose of the Study:

• To develop a highly sensitive signal-amplified platform for detecting IFN-γ.
• To integrate aptamer recognition, strand displacement amplification (SDA), CRISPR/Cas12a, and cobalt oxyhydroxide nanosheets for enhanced detection.

Main Methods:

• Aptamer-target IFN-γ binding initiated cDNA release from an aptamer-complementary DNA duplex.
• Released cDNA triggered SDA, generating double-stranded DNA products.
• Activated CRISPR/Cas12a cleaved a FAM-labeled DNA probe, enabling fluorescence recovery for detection.

Main Results:

• The Apt-SCN tandem system achieved sensitive fluorometric analysis of IFN-γ.
• A low detection limit of 0.37 nM for IFN-γ was established.
• The system demonstrated potential for detecting other cytokines.

Conclusions:

• The Apt-SCN tandem system offers a novel, multi-amplification approach for IFN-γ detection.
• This is the first study to use SDA and CRISPR/Cas12a for IFN-γ detection.
• The system shows significant potential for clinical cytokine assays and CS prevention.