Nucleic Acid Nanotechnology-Empowered CRISPR-Cas12a Systems for Biosensing and Bioimaging Applications.

The development of highly sensitive and simple bioanalytical platforms is crucial for advancing disease diagnostics and biomedical research. In recent years, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated (Cas) protein systems have emerged as the "next-generation molecular technology" that profoundly impacts the field of genome editing and molecular diagnosis. In particular, CRISPR-Cas12a, an important CRISPR-Cas family member, has gained prominence as a robust tool in biosensing and bioimaging due to its easy design and high target specificity. Notably, the integration of nucleic acid nanotechnology, including nucleic acid amplification strategies and engineered functional nucleic acids, with CRISPR-Cas12a systems significantly improves detection sensitivity and specificity, enabling the analysis of low-abundance analytes. This review introduces the fundamentals of CRISPR-Cas12a and key nucleic acid-based toolboxes. Next, we systematically summarize the advantages of nucleic acid nanotechnology-empowered CRISPR-Cas12a platforms in detail and highlight recent advances in their applications in biosensing and bioimaging. Finally, current challenges and future perspectives of such nucleic acids-assisted CRISPR-Cas12a systems for disease diagnostics are discussed.