Three-Way Junction-Assisted Rolling Circle Amplification Integrated with <i>trans-</i>Cleavage of Cas12a for Sensitive and Reliable Detection of miRNA

作者全名："Wu, Zhiquan; Zhao, Shuqi"

作者地址："[Wu, Zhiquan] Chongqing Med Univ, Emergency & Crit Care Ctr, Affiliated Hosp 3, Chongqing 401120, Peoples R China; [Zhao, Shuqi] Chongqing Med Univ, Dept Hematol, Affiliated Hosp 3, Shuanghu Branch Rd, Chongqing 401120, Peoples R China"

通信作者："Zhao, SQ (通讯作者)，Chongqing Med Univ, Dept Hematol, Affiliated Hosp 3, Shuanghu Branch Rd, Chongqing 401120, Peoples R China."

来源：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY

ESI学科分类：BIOLOGY & BIOCHEMISTRY

WOS号：WOS:001063566500005

JCR分区：Q2

影响因子：3.1

年份：2023

卷号：　

期号：　

开始页：　

结束页：　

文献类型：Article; Early Access

关键词：MicroRNAs (miRNAs); Rolling circle amplification (RCA); Catalytic hairpin amplification (CHA); Cas12a

摘要："MicroRNAs (miRNAs) serve a crucial role in numerous biological processes, such as acute pancreatitis development. Due to its low abundance and high similarity among homogeneous family members, sensitive and reliable detection of microRNA remains a formidable challenge. By combining the three-way junction-assisted rolling circle amplification (RCA) with the trans-cleavage of Cas12a, we propose a novel fluorescent technique for sensitive miRNA detection. In order to increase the amplification efficiency of RCA-based methods, catalytic hairpin amplification (CHA) is incorporated into the RCA process, playing the roles of specific target recognition and three-way junction formation. Consequently, the method demonstrated a six-orders-of-magnitude detection range and a LOD as low as 27 aM, making it a promising method for the early diagnosis of various diseases."

基金机构：We appreciate the financial support from the Chongqing Medical University. We also thank the director of central laboratory for providing essential equipment.; Chongqing Medical University

基金资助正文：We appreciate the financial support from the Chongqing Medical University. We also thank the director of central laboratory for providing essential equipment.