School of Chemistry and Materials Science, Jiangsu Normal University, Xuzhou 221116, China.
Guangzhou Double Helix Gene Technology Co., Ltd., Guangzhou International Bio Island Co., Ltd., Guangzhou 510320, P.R. China.
Anal Chem. 2020 Mar 3;92(5):4029-4037. doi: 10.1021/acs.analchem.9b05597. Epub 2020 Feb 17.
Gold-nanoparticles-based colorimetric assay is an attractive detection format, but is limited by the tedious and ineffective posthybridization manipulations for genomic analysis. Here, we present a new design for a colorimetric gene-sensing platform based on the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system. In this strategy, programmable recognition of DNA by Cas12a/crRNA and RNA by Cas13a/crRNA with a complementary target activates the -ssDNA or -ssRNA cleavage. Target-induced -ssDNA or ssRNA cleavage triggers an aggregation behavior change for the designed AuNPs-DNA probes pair, enabling the completion of naked-eye gene detection (transgenic rice, African swine fever virus, and miRNAs as the models) within 1 h. This platform is also showing promise as a fast and inexpensive tool for bacteria identification using or . A CRISPR/Cas-based colorimetric platform shows superior characteristics, such as probe universality, compatibility with isothermal reaction conditions, on-site detection capability, and high sensitivity, thus, demonstrating its use as a robust next-generation gene detection platform.
基于金纳米粒子的比色分析是一种很有吸引力的检测形式，但由于基因组分析中杂交后繁琐且低效的操作而受到限制。在这里，我们提出了一种基于成簇规律间隔短回文重复序列（CRISPR）/Cas 系统的新型比色基因传感平台设计。在该策略中，Cas12a/crRNA 对 DNA 的可编程识别和 Cas13a/crRNA 对互补靶标的 RNA 的识别激活了 -ssDNA 或 -ssRNA 的切割。靶标诱导的 -ssDNA 或 ssRNA 切割引发设计的 AuNPs-DNA 探针对的聚集行为变化，从而在 1 小时内完成肉眼可见的基因检测（以转基因水稻、非洲猪瘟病毒和 miRNA 为模型）。该平台还显示出作为一种快速且廉价的细菌鉴定工具的潜力，可用于 或 。基于 CRISPR/Cas 的比色平台具有探针通用性、与等温反应条件的兼容性、现场检测能力和高灵敏度等优越特性，因此可作为一种强大的下一代基因检测平台。
