CRISPR/cas systems redefine nucleic acid detection: Principles and methods
CRISPR/cas systems redefine nucleic acid detection: Principles and methods
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DOI：
10.1016/j.bios.2020.112430
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发表时间：
2020-10-01
影响因子：
12.6
通讯作者：
Li, Jinming
中科院分区：
文献类型：
--
作者：
Wang, Meng;Zhang, Rui;Li, Jinming
能够对核酸序列进行快速、灵敏和特异性分析的方法通过提供对临床相关遗传信息的直接洞察，对精确的疾病诊断和有效的临床治疗具有积极影响。到目前为止，许多CRISPR/Cas系统已经被重新用于诊断功能，并且由于其高度的灵活性，灵敏度和特异性，正在彻底改变强大诊断工具的可访问性。作为RNA引导的靶向识别效应物，Cas9变体已用于各种诊断应用，包括生物传感测定、成像测定和用于下一代测序（NGS）的靶富集，从而能够开发灵活且具有成本效益的测试。此外，随后发现的Cas蛋白（Cas 12和Cas 13）具有侧切活性，促进了许多诊断工具的发展，用于快速和便携式检测，这些工具具有很大的潜力，为现场护理设置。然而，在这个问题上提出的代表性评论主要局限于经典的生物传感应用，因此，这个快速发展的领域需要一个全面和系统的描述。在这篇综述中，基于检测原理，我们从一个新的角度对最近利用这些基于CRISPR的诊断工具的工作进行了详细的分类和全面的讨论。此外，还概述了基于CRISPR的诊断的当前挑战和未来前景。
Methods that enable rapid, sensitive and specific analyses of nucleic acid sequences have positive effects on precise disease diagnostics and effective clinical treatments by providing direct insight into clinically relevant genetic information. Thus far, many CRISPR/Cas systems have been repurposed for diagnostic functions and are revolutionizing the accessibility of robust diagnostic tools due to their high flexibility, sensitivity and specificity. As RNA-guided targeted recognition effectors, Cas9 variants have been utilized for a variety of diagnostic applications, including biosensing assays, imaging assays and target enrichment for next-generation sequencing (NGS), thereby enabling the development of flexible and cost-effective tests. In addition, the ensuing discovery of Cas proteins (Cas12 and Cas13) with collateral cleavage activities has facilitated the development of numerous diagnostic tools for rapid and portable detection, and these tools have great potential for point-of-care settings. However, representative reviews proposed on this topic are mainly confined to classical biosensing applications; thus, a comprehensive and systematic description of this fast-developing field is required. In this review, based on the detection principle, we provide a detailed classification and comprehensive discussion of recent works that harness these CRISPR-based diagnostic tools from a new perspective. Furthermore, current challenges and future perspectives of CRISPR-based diagnostics are outlined.