CRISPR/Cas系统在生物核酸检测中的应用进展

Advances in application of CRISPR/Cas system in biological nucleic acid detection

传统的核酸检测技术耗时长且对操作人员的专业水平要求高,开发精准、快速、便携、低成本、高特异性和灵敏度的新核酸检测系统具有重要意义。目前,CRISPR/Cas系统不但被广泛应用于基因编辑,而且随着部分Cas蛋白“附带切割”活性的发现,灵敏度高、特异性强的CRISPR/Cas系统被逐步开发为新型生物传感工具,用于生物的核酸检测。随着各种Cas效应蛋白的发现,研究人员开发了多种基于CRISPR/Cas系统的核酸检测方法(如SHERLOCK、HOLMES、DETECTR、HUDSON和CASLFA等)可检测各种靶标(包括细菌、病毒、癌症突变等)。笔者综述了当前重要的用于不同靶标检测的CRISPR/Cas系统,包括检测核酸、其他小分子物质及病原微生物,讨论了其面临的挑战和应用潜力,最后推断,随着CRISPR/Cas系统的不断发展和完善,该系统在生物传感及临床分子检测领域的应用会越来越广泛。

Traditional nucleic acid detection technology is time-consuming and requires high professional level of operators.Therefore,it is of great significance to develop a new nucleic acid detection system that is accurate,fast,portable,low-cost,and highly specific and sensitive.At present,CRISPR/Cas is not only widely used in gene editing,but with the discovery of the"incidental cleavage"activity of some Cas proteins.CRISPR/Cas system with high sensitivity and strong specificity has been gradually developed as a new type of biosensing tool for nucleic acid detection of organisms.With the emergence of various Cas effector proteins,researchers have developed a variety of nucleic acid assessors based on the CRISPR/Cas system(SHERLOCK,HOLMES,DETECTR,HUDSON,CASLFA,etc.)to detect various targets(including bacteria,viruses,cancer mutations and more).We review the current important CRISPR/Cas systems for the detection of different targets,including the detection of nucleic acids,other small molecules,and pathogenic microorganisms,and discuss the challenges they face and their application potential.Finally,we conclude that with the continuous development and improvement of the CRISPR/Cas system,the system will be more and more widely used in the fields of biosensing and clinical molecular detection.