School of Life Sciences, South China Normal University, Guangzhou, China; email:
Annu Rev Anal Chem (Palo Alto Calif). 2023 Jun 14;16(1):311-332. doi: 10.1146/annurev-anchem-090822-014725. Epub 2023 Apr 5.
Biosensing based on CRISPR-Cas systems is a young but rapidly evolving technology. The unprecedented properties of the CRISPR-Cas system provide an innovative tool for developing new-generation biosensing strategies. To date, a series of nucleic acid and non-nucleic acid detection methods have been developed based on the CRISPR platform. In this review, we first introduce the core biochemical properties underpinning the development of CRISPR bioassays, such as diverse reaction temperatures, programmability in design, high reaction efficiency, and recognition specificity, and highlight recent efforts to improve these parameters. We then introduce the technical developments, including how to improve sensitivity and quantification capabilities, develop multiplex assays, achieve convenient one-pot assays, create advanced sensors, and extend the applications of detection. Finally, we analyze obstacles to the commercial application of CRISPR detection technology and explore development opportunities and directions.
基于 CRISPR-Cas 系统的生物传感是一项年轻但快速发展的技术。CRISPR-Cas 系统前所未有的特性为开发新一代生物传感策略提供了创新工具。迄今为止,已经基于 CRISPR 平台开发了一系列核酸和非核酸检测方法。在这篇综述中,我们首先介绍了支撑 CRISPR 生物分析发展的核心生化特性,例如多样化的反应温度、设计上的可编程性、高反应效率和识别特异性,并强调了最近为改善这些参数所做的努力。然后,我们介绍了技术发展,包括如何提高灵敏度和定量能力、开发多重分析、实现方便的一键式分析、创建先进的传感器以及扩展检测应用。最后,我们分析了 CRISPR 检测技术商业化应用的障碍,并探讨了发展机遇和方向。