Yi Bo, Zhou Baoqing, Zhou Donggen, Yang Luyu, Xu Hengyi
State Key Laboratory of Food Science and Resources, Nanchang University, Nanchang 330047, PR China.
Jiangxi General Institute of Testing and Certification, Nanchang 330052, PR China.
Biotechnol Adv. 2025 Oct;83:108609. doi: 10.1016/j.biotechadv.2025.108609. Epub 2025 May 21.
Rapid, accurate, cost-effective, and efficient ultrasensitive detection strategies are essential for public health safety (including food safety, disease prevention and environmental governance). The CRISPR/CRISPR-associated (Cas) detection is a cutting-edge technology that has been widely used in the detection of public health safety due to its targeted cleavage properties (signal amplification), attomolar level sensitivity, high specificity (recognizing single-base mismatches), and rapid turnover time. However, the current research about CRISPR/Cas-based biosensors is not clear, such as mechanism problem and application differences of integrating CRISPR/Cas system with other technologies, and how to further innovate and develop in the future. Therefore, further detailed analysis and comparative discussion of CRISPR/Cas-based biosensors is needed. Currently, CRISPR/Cas system powered biosensors can be mainly categorized into two types: CRISPR/Cas system powered nucleic acid amplification biosensors and CRISPR/Cas system powered nucleic acid amplification-free biosensors. The two biosensors have different characteristics and advantages. This paper first provides an in-depth investigation of the enzymatic mechanism of CRISPR/Cas system at the molecular level. Then, this paper summarizes the principles and recent advances of CRISPR/Cas system powered nucleic acid amplification biosensors and CRISPR/Cas system powered nucleic acid amplification-free biosensors and discusses their integration mechanisms in depth. More, the differences and application-oriented between the two biosensors are further discussed. Finally, the application orientation and future perspectives of the two biosensors are discussed, and unique insights into the future development of CRISPR/Cas system are provided.
快速、准确、经济高效且高效的超灵敏检测策略对于公共卫生安全（包括食品安全、疾病预防和环境治理）至关重要。CRISPR/CRISPR相关（Cas）检测是一项前沿技术，由于其靶向切割特性（信号放大）、阿托摩尔级灵敏度、高特异性（识别单碱基错配）和快速周转时间，已被广泛应用于公共卫生安全检测。然而，目前关于基于CRISPR/Cas的生物传感器的研究尚不清楚，例如将CRISPR/Cas系统与其他技术整合的机制问题和应用差异，以及未来如何进一步创新和发展。因此，需要对基于CRISPR/Cas的生物传感器进行更详细的分析和比较讨论。目前，由CRISPR/Cas系统驱动的生物传感器主要可分为两类：由CRISPR/Cas系统驱动的核酸扩增生物传感器和由CRISPR/Cas系统驱动的无核酸扩增生物传感器。这两种生物传感器具有不同的特点和优势。本文首先在分子水平上深入研究了CRISPR/Cas系统的酶促机制。然后，本文总结了由CRISPR/Cas系统驱动的核酸扩增生物传感器和由CRISPR/Cas系统驱动的无核酸扩增生物传感器的原理和最新进展，并深入讨论了它们的整合机制。此外，进一步讨论了这两种生物传感器之间的差异和应用导向。最后，讨论了这两种生物传感器的应用方向和未来前景，并对CRISPR/Cas系统的未来发展提供了独特见解。
