Aptamer-Linked CRISPR/Cas12a-Based Immunoassay

Area of Science:

• Biotechnology
• Molecular Biology
• Biosensing

Background:

• The CRISPR/Cas system shows potential in nucleic acid biosensing.
• Current limitations include low versatility, moderate sensitivity, and complex operations for non-nucleic acid targets, hindering clinical translation.

Purpose of the Study:

• To develop a versatile and sensitive detection method for non-nucleic acid analytes using the CRISPR/Cas12a system.
• To overcome the limitations of existing CRISPR-based biosensing platforms for clinical diagnostics.

Main Methods:

• Designed aptamer-flanked activator DNA strands to link non-nucleic acid analytes to the CRISPR/Cas12a system.
• Utilized the collateral cleavage activity of CRISPR/Cas12a for signal amplification.
• Adapted the sensing principle to be compatible with existing enzyme-linked immunosorbent assay (ELISA) platforms.

Main Results:

• Achieved ultrasensitive detection of biomarkers from various species.
• Demonstrated a direct correlation between non-nucleic acid analytes and CRISPR/Cas12a activation.
• The system offers optional signal output and compatibility with ELISA platforms.

Conclusions:

• The developed method significantly broadens the applicability of CRISPR/Cas systems in bioanalysis and medical diagnostics.
• This approach facilitates the clinical transition of CRISPR-based biosensing technologies by enhancing sensitivity, versatility, and ease of use.