Karimi Marzieh, Ghorbani Abozar, Niazi Ali, Rostami Mahsa, Tahmasebi Ahmad
Institute of Biotechnology, Shiraz University, Shiraz, Iran.
Nuclear Agriculture Research School, Nuclear Science and Technology Research Institute (NSTRI), Karaj, Iran.
Plant Methods. 2025 Jun 9;21(1):83. doi: 10.1186/s13007-025-01401-9.
Plant viruses are among the most serious threats to global agriculture, causing significant yield losses and jeopardizing food security. Identifying these viruses is crucial to prevent widespread crop damage and ensure effective management. CRISPR-Cas13a, a subtype of the RNA-targeting Cas13 family, has emerged as a transformative tool in molecular diagnostics, specifically tailored to detect these plant RNA viruses with unparalleled precision. Unlike traditional methods such as ELISA and RT-PCR, which are often limited by sensitivity, equipment dependency, and long processing times, Cas13a offers exceptional specificity and attomolar-level sensitivity. Its RNA-guided collateral cleavage mechanism allows signal amplification, making it particularly suitable for field-deployable diagnostics. Recent advances in Cas13 engineering, including compact variants such as Cas13bt3 and Cas13Y, have further improved its delivery efficiency and minimized immune responses, enhancing its agricultural applications. Integration with amplification methods like LAMP and innovative biosensor platforms like graphene-based and electrochemical systems further enhances its diagnostic potential. While challenges remain, including off-target effects, reagent stability, and scalability, innovations in CRISPR RNA (crRNA) design, reagent encapsulation, and microfluidic technologies are actively addressing these barriers. CRISPR-Cas13a represents a cutting-edge solution for rapid, accurate, and accessible plant virus diagnostics, providing a powerful safeguard for crop yields and global food security.
植物病毒是全球农业面临的最严重威胁之一，会导致严重的产量损失并危及粮食安全。识别这些病毒对于防止作物广泛受损并确保有效管理至关重要。CRISPR-Cas13a是RNA靶向Cas13家族的一个亚型，已成为分子诊断中的一种变革性工具，专门用于以无与伦比的精度检测这些植物RNA病毒。与酶联免疫吸附测定（ELISA）和逆转录聚合酶链反应（RT-PCR）等传统方法不同，传统方法往往受到灵敏度、设备依赖性和处理时间长的限制，Cas13a具有卓越的特异性和阿托摩尔级别的灵敏度。其RNA引导的附带切割机制允许信号放大，使其特别适用于可现场部署的诊断。Cas13工程的最新进展，包括Cas13bt3和Cas13Y等紧凑型变体，进一步提高了其递送效率并将免疫反应降至最低，增强了其在农业中的应用。与环介导等温扩增（LAMP）等扩增方法以及基于石墨烯和电化学系统等创新生物传感器平台的整合进一步增强了其诊断潜力。虽然挑战仍然存在，包括脱靶效应、试剂稳定性和可扩展性，但CRISPR RNA（crRNA）设计、试剂封装和微流控技术方面的创新正在积极应对这些障碍。CRISPR-Cas13a代表了一种用于快速、准确和便捷的植物病毒诊断的前沿解决方案，为作物产量和全球粮食安全提供了有力保障。
