摘　要：近年来，病毒感染疫情频发，凸显了高效便利的病毒检测技术以及抗病毒药物研制的迫切性。基于成簇的规则间隔短回文重复序列（CRISPR）和CRISPR相关蛋白（Cas）的工程系统在靶向和切割核酸方面具有较高的特异性和效率，是目前使用最为广泛的基因编辑工具。该系统目前也广泛应用于病毒学研究和相关医疗实践。本文重点介绍了Cas9、Cas12和Cas13这三种最常用的CRISPR/Cas系统在病毒检测和抗病毒治疗中的应用。在病毒检测方面，Cas9通过与荧光传感器、电化学传感器和侧流层析试纸等生物传感器相结合，提高了生物传感器检测的灵敏度和准确性。Cas12和Cas13则基于其反式切割活性，目前已经开发了多种技术来检测DNA和RNA病毒，如SHERLOCK和DETECTER。在抗病毒治疗方面，Cas9已被用于靶向切割病毒DNA，从而抑制病毒的复制，其靶标包括DNA病毒的基因组和逆转录病毒的中间产物DNA；而Cas13则被用于靶向病毒RNA，其靶标包括RNA病毒的基因组和病毒mRNA。尽管CRISPR/Cas系统在灵敏度、效率和便利度等方面具有多种优势，但在一些方面仍不可避免地存在局限性，如脱靶效应、免疫原性和致癌性。本文全面总结了CRISPR/Cas系统应用于病毒检测和抗病毒治疗的现有进展，为了解该领域相关技术提供了系统的参考。

关键词：CRISPR/Cas系统；病毒检测；抗病毒治疗；生物传感器

中图分类号：Q789                             文献标志码：A              DOI：10.3969/j.issn.1007-7146.2023.06.003

（Acta Laser Biology Sinica, 2023, 32(6): 502-516）

Abstract: Frequent viral epidemics in recent years urge the development of efficient and convenient techniques for virus detection and antiviral drugs development. As the most popular gene-editing tools, engineered systems based on clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) possess high specificity and efficiency in nucleic acid targeting and cleavage, which have also been widely used in virological research and medical practice. This review briefly introduces the features of three most commonly used CRISPR/Cas systems, including Cas9, Cas12 and Cas13, and comprehensively concludes their application in virus detection and antiviral therapeutics. For virus detection, Cas9 enhances the sensitivity and accuracy of biosensor detection by combining it with biosensors such as fluorescent sensors, electrochemical sensors, and lateral flow chromatography. As for Cas12 and Cas13, multiple technologies have been developed to detect DNA and RNA viruses based on their trans-cleavage activity, such as SHERLOCK and DETECTR. As CRISPR/Cas-based antiviral techniques, Cas9 has been employed to cleave viral DNA, including genomes of DNA viruses and intermediate DNA of retroviruses, while Cas13 has been used to target viral RNA, including genomes of RNA viruses and viral mRNA. Although CRISPR/Cas systems have shown multiple advantages in sensitivity, efficiency and convenience, they still bear some limitations, such as off-target effect, immunogenicity and carcinogenicity. In summary, this manuscript provides an overview of current progress in the application of different CRISPR/Cas systems as promising tools in virus detection and antiviral therapy.

Key words: CRISPR/Cas system; virus detection; antiviral therapy; biosensor

CLC number: Q789                        Document code: A                DOI：10.3969/j.issn.1007-7146.2023.06.003