CRISPR-Cas技术在SARS-CoV-2核酸检测中的研究进展

Research progress of CRISPR-CAS system in SARS-CoV-2 nucleic acid detection

规律成簇的间隔短回文重复序列及其相关蛋白[clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas), CRISPR-Cas]组成原核生物的适应性免疫系统。CRISPR-Cas核酸检测技术主要依赖于crRNA和靶标序列识别结合后激发Cas蛋白的核酸探针切割活性。将CRISPR-Cas系统应用于核酸检测中,为提高核酸检测的灵敏度和特异度提供了一种新方法。目前,已有多项基于CRISPR-Cas系统的核酸检测研究,且检测性能优越、简便、快速,有广泛的应用价值。现将基于第二类CRISPR-Cas技术的严重急性呼吸综合征冠状病毒2(severe acute respiratory syndrome-coronavirus 2, SARS-CoV-2)核酸检测新方法作一概述,为后期开发的基于CRISPR-Cas针对SARS-CoV-2变异毒株单碱基突变核酸检测技术提供参考和依据。

CRISPR-Cas system is a prokaryotic adaptive immune system composed of clustered regularly interspaced short palindromic repeats(CRISPR) and CRISPR-associated proteins(Cas). CRISPR-Cas technologies mainly depend on the cleavage activity of Cas proteins after the recognition and combination of crRNA and target sequence. The application of CRISPR-Cas system in nucleic acid detection provides a novel method to improve the sensitivity and specificity of detection. At present, many nucleic acid detection studies based on CRISPR-Cas system have been reported and confirm superiority performance and extensive application value of this simple rapid method. This paper summarizes the SARS-CoV-2 nucleic acid detection technologies based on the Class II CRISPR-Cas, to provide reference and basis for future development of CRISPR-Cas based single base mutations detection for SARS-CoV-2 variants.