Biochemistry

The Emerging Improved Approaches for Crispr/cas-Based Detection: Recent Advance and Perspectives

Authors: Ziyue Liang, Xinran Xiang
Comments: 44 Pages.

The CRISPR/Cas system has emerged as a revolutionary tool for nucleic acid detection, offering high sensitivity, specificity, and versatility. This review highlights recent advancements in CRISPR/Cas-based detection technologies, focusing on structural improvements to key components such as crRNA, Cas proteins, PAM sequences, probes, and activators. These enhancements have significantly boosted the system's efficiency, accuracy, and applicability. Furthermore, the integration of CRISPR/Cas with nucleic acid amplification techniques (e.g., PCR, LAMP, RPA, SDA), microfluidic platforms, nanomaterials, and biosensors (fluorescence, colorimetric, electrochemical, and lateral flow) has enabled rapid, portable, and cost-effective diagnostics. These innovations address critical challenges in infectious disease control, food safety, and clinical diagnostics, particularly in resource-limited settings. Despite these advancements, limitations such as PAM/PFS restrictions, off-target effects, and the need for faster reaction times remain. Future directions include exploring novel Cas enzymes, optimizing reaction conditions, and developing anti-CRISPR strategies to further refine this transformative technology. This review provides a comprehensive overview of the current state and future prospects of CRISPR/Cas-based detection systems, underscoring their potential to revolutionize point-of-care diagnostics and large-scale screening.
Category: Biochemistry