Novel strategies for biomolecular detection by utilizing the trans-cleavage activity of CRISPR/Cas system

Abstract

Since the outbreak of the COVID-19 pandemic, the importance of molecular diagnostics has emerged and the related market has grown exponentially. Nucleic acids have been widely used as a major target of such molecular diagnosis. Detection of nucleic acids is very important for accurate and rapid diagnosis and effective control of infectious diseases. CRISPR/Cas system is a type of adaptive immunity of microorganisms, an immune system that bacteria have evolved to protect themselves from viral infection. Recently, CRISPR/Cas system is extensively applied to the field of molecular diagnosis beyond genome editing thanks to its high target- specificity and rapid kinetics. The applicability of CRISPR/Cas system has also been tested for the detection of various biomolecules. In this study, in order to utilize CRISPR/Cas system for the development of various biomolecules detection technologies and molecular diagnosis technologies, CRISPR/Cas12a system was applied for the detection of ribonuclease H, protein-small molecule interactions, and target nucleic acids. The developed technologies are expected to have great social and economic ramifications in the field of in vitro diagnostics in the future, considering their excellent sensitivity, selectivity, and applicability to biological samples.