文献类型：期刊文献

英文题名：A programmable and sensitive CRISPR/Cas12a-based MicroRNA detection platform combined with hybridization chain reaction

作者：Jia, Hai-Yan[1];Zhao, Hong-Li[1];Wang, Ting[1];Chen, Pin-Ru[2];Yin, Bin-Cheng[1,2,3];Ye, Bang-Ce[1,2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Lab Biosyst & Microanal, Shanghai 200237, Peoples R China;[2]Zhejiang Univ Technol, Coll Pharmaceut Sci, Collaborat Innovat Ctr Yangtze River Delta Reg Gre, Hangzhou 310014, Zhejiang, Peoples R China;[3]Shihezi Univ, Sch Chem & Chem Engn, Shihezi 832000, Xinjiang, Peoples R China

年份：2022

卷号：211

外文期刊名：BIOSENSORS & BIOELECTRONICS

收录：;EI(收录号：20222112147276);WOS:【SCI-EXPANDED(收录号:WOS:000806387700003)】；

基金：Acknowledgments This work was jointly supported by the National Natural Science Foundation of China (Grants 21822402, 31730004) , the National Key R&D Program of China (Grant 2020YFA0908800) , ?Shuguang Program? supported by Shanghai Education Development Foundation and Shanghai Municipal Education Commission, and the Joint Funds of the Zhejiang Provincial Natural Science Foundation of China (Grant LHDMZ22H300008) .

语种：英文

外文关键词：MicroRNA detection; CRISPR; Cas; Collateral cleavage; Hybridization chain reaction circuit; Signal amplification

摘要：MicroRNAs (miRNAs) play an essential role in cancer diagnosis and prognosis. Developing a new method for sensitive detection of miRNA is constantly in demand. CRISPR/Cas12a system can nonspecifically cleave singlestranded DNA after specific recognition of target DNA, showing tremendous potential in molecular diagnostics. However, CRISPR-based detection methods require synthesizing different crRNAs for detecting different targets, which limit their widespread application. Herein, we design a versatile and sensitive miRNA detection platform based on CRISPR/Cas12a system combined with a hybridization chain reaction (HCR) circuit. In this design, the HCR circuit as the signal transducer converts each miRNA into multiple DNA duplexes, which act as the activators to activate the trans-cleavage activity of Cas12a for further signal amplification. More importantly, this platform can sensitively detect different miRNAs without changing the spacer sequence of crRNA due to the fixed activators formed by HCR. In addition, the consistency between the proposed platform and RT-qPCR in miRNA detection extracted from different cell lines validated its practicability, demonstrating the potential in clinical diagnosis of cancers and monitoring therapy.