Wei Hongguo, Bu Shengjun, Wang Ze, Zhou Hongyu, Li Xue, Wei Jiaqi, He Xiuxia, Wan Jiayu
School of Life Science and Technology, Changchun University of Science and Technology, Changchun 130022, China.
Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun 130122, China.
ACS Omega. 2022 Sep 26;7(40):35515-35522. doi: 10.1021/acsomega.2c01930. eCollection 2022 Oct 11.
MicroRNAs (miRNAs) play a very important role in biological processes and are used as biomarkers for the detection of a variety of diseases, including neurodegenerative diseases, chronic cardiovascular diseases, and cancers. A sensitive point-of-care (POC) method is crucial for detecting miRNAs. Herein, CRISPR-Cas12a combined with the click chemistry actuated exponential amplification reaction was introduced into an electrochemical biosensor for detecting miRNA-21. The target miRNA-21 initiated the click chemistry-exponential amplification reaction in the electrochemical biosensor to produce numerous nucleic acid fragments, which could stimulate the trans-cleavage ability of CRISPR-Cas12a to cleave hairpin DNA electrochemical reporters immobilized on the electrode surface. Under optimal conditions, the minimum detection limit for this electrochemical biosensor was as low as 1 fM. Thus, the proposed electrochemical biosensor allows sensitive and efficient miRNA detection and could be a potential analysis tool for POC test and field molecular diagnostics.
微小RNA（miRNA）在生物过程中发挥着非常重要的作用，并被用作检测多种疾病的生物标志物，包括神经退行性疾病、慢性心血管疾病和癌症。一种灵敏的即时检测（POC）方法对于检测miRNA至关重要。在此，将CRISPR-Cas12a与点击化学驱动的指数扩增反应相结合引入到一种用于检测miRNA-21的电化学生物传感器中。目标miRNA-21在电化学生物传感器中引发点击化学-指数扩增反应，产生大量核酸片段，这些片段可刺激CRISPR-Cas12a的反式切割能力，以切割固定在电极表面的发夹状DNA电化学报告分子。在最佳条件下，这种电化学生物传感器的最低检测限低至1 fM。因此，所提出的电化学生物传感器能够灵敏且高效地检测miRNA，并且可能成为POC检测和现场分子诊断的潜在分析工具。
