Research on the Biosensing Mechanisms of CRISPR/Cas9 Systems and the Applications in Detection of Non-Infectious Disease

Abstract

The CRISPR/Cas9 system has become a groundbreaking technology in molecular diagnostics due to its precise nucleic acid recognition capabilities. This paper comprehensively analyzes the molecular mechanisms of CRISPR-based biosensing platforms, emphasizing target specificity and signal transduction. It reveals how engineered Cas9 variants and optimized guide RNA enhance detection sensitivity, with applications in non-infectious disease monitoring (e.g., early cancer biomarkers). CRISPR-mediated platforms achieve remarkable performance, with detection limits at femtomolar concentrations. The review discusses challenges like off-target effects and signal amplification, proposing solutions involving nanotechnology and microfluidics. Chronic non-communicable conditions challenge global healthcare. CRISPR/Cas9 comprises gRNA and Cas9, with class 2 systems (e.g., Type II) prominent. Cas9 identifies sequences via protospacer adjacent motifs, inducing DNA breaks. Traditional methods are labor-intensive; CRISPR Class 2 platforms enable sensitive, precise detection at lower costs. Emerging Cas variants (e.g., Cpf1) offer enhanced capabilities. Challenges include sample processing and non-specific binding. Future research should simplify protocols and conduct clinical trials. CRISPR/Cas9 advances precision genetic analysis.