Abstract：
An electrochemical sensor assisted by primer exchange reaction (PER) and CRISPR/Cas9 system (PER-CRISPR/Cas9-E) was established for the sensitive detection of dual microRNAs (miRNAs). Two PER hairpin (HP) were designed to produce a lot of extended PER products, which could hybridize with two kinds of hairpin probes modified on the electrode and initiate the cleavage of two CRISPR/Cas9 systems guided by single guide RNAs (sgRNAs) with different recognition sequences. The decrease of the two electrochemical redox signals indicated the presence of dual-target miRNAs. With the robustness and high specificity of PER amplification and CRISPR/Cas9 cleavage system, simultaneous detection of two targets was achieved and the detection limits for miRNA-21 and miRNA-155 were 0.43 fM and 0.12 fM, respectively. The developed biosensor has the advantages of low cost, easy operation, and in-situ detection, providing a promising platform for point-of-care detection of multiple miRNAs.
摘要：
建立了基于引物交换反应（PER）和CRISPR/Cas9系统（PER-CRISPR/Cas9-E）的电化学传感器，用于敏感检测双microRNAs（miRNAs）。设计了两个PER发夹（HP）来生产许多扩展的PER产品，它可以与电极上修饰的两种发夹探针杂交，并启动由具有不同识别序列的单向导RNA（sgRNA）引导的两个CRISPR/Cas9系统的裂解。两个电化学氧化还原信号的降低表明存在双靶标miRNA。PER扩增和CRISPR/Cas9裂解系统的鲁棒性和高特异性,实现了两个目标的同时检测，miRNA-21和miRNA-155的检出限分别为0.43fM和0.12fM，分别。所研制的生物传感器具有成本低,操作简单,和原位检测，为多种miRNA的即时检测提供了一个有前途的平台。
