New CRISPR-Derived microRNA Sensing Mechanism Based on Cas12a Self-Powered and Rolling Circle Transcription-Unleashed Real-Time crRNA Recruiting

被引：111

作者：

Wang, Gaoting ^{[1
,2
]}

Tian, Weimin ^{[1
,2
]}

Liu, Xiaoling ^{[1
,2
]}

Ren, Wei ^{[1
,2
]}

Liu, Chenghui ^{[1
,2
]}

机构：

[1] Shaanxi Normal Univ, Key Lab Appl Surface & Colloid Chem, Minist Educ, Xian 710119, Shaanxi, Peoples R China

[2] Shaanxi Normal Univ, Sch Chem & Chem Engn, Key Lab Analyt Chem Life Sci Shaanxi Prov, Xian 710119, Shaanxi, Peoples R China

来源：

ANALYTICAL CHEMISTRY | 2020年 / 92卷 / 09期

基金：

中国国家自然科学基金;

关键词：

RNA-GUIDED ENDONUCLEASE; CATALYTIC RNAS; DNA; AMPLIFICATION; DIAGNOSTICS; POLYMERASE; GENERATION; ASSAY; CPF1;

D O I：

10.1021/acs.analchem.0c00680

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Current CRISPR-Cas-based nucleic acid sensing methods relying on the preassembled Cas-crRNA complexes are generally limited to the detection of protospacer-adjacent motif (PAM)-containing sequences, and nonspecific backgrounds are inevitable. Herein, we propose a new CRISPR-derived microRNA sensing mechanism based on rolling circle transcription (RCT)-unleashed self-recruiting of crRNA by Cas12a (Cas12a-SCR). In Cas12a-SCR, target microRNA can specifically trigger RCT to produce a long single- strand RNA with numerous pre-crRNA repeats, which can be trimmed and recruited by Cas12a actively. This new target-initiated, real-time producing, trimming, and self-assembling manner of Cas12a-crRNA remarkably suppresses the nonspecific background and relieves the stringent requirement of PAM site in the target sequence. Thus, the universality of the Cas12a-SCR toward different nucleic acid sequences is greatly expanded.

引用

页码：6702 / 6708

页数：7

共 55 条

[1] CRISPR/Cas13a-Powered Electrochemical Microfluidic Biosensor for Nucleic Acid Amplification-Free miRNA Diagnostics [J].