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

Hai-Yan Jia; Hong-Li Zhao; Ting Wang; Pin-Ru Chen; Bin-Cheng Yin; Bang-Ce Ye

doi:10.1016/j.bios.2022.114382

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

Biosens Bioelectron. 2022 Sep 1:211:114382. doi: 10.1016/j.bios.2022.114382. Epub 2022 May 18.

Authors

Hai-Yan Jia¹, Hong-Li Zhao¹, Ting Wang¹, Pin-Ru Chen², Bin-Cheng Yin³, Bang-Ce Ye⁴

Affiliations

¹ Lab of Biosystem and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.
² Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, Zhejiang, China.
³ Lab of Biosystem and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China; Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, Zhejiang, China; School of Chemistry and Chemical Engineering, Shihezi University, Xinjiang, 832000, China. Electronic address: binchengyin@ecust.edu.cn.
⁴ Lab of Biosystem and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China; Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, Zhejiang, China.

PMID: 35605545
DOI: 10.1016/j.bios.2022.114382

Abstract

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 single-stranded 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.

Keywords: CRISPR/Cas; Collateral cleavage; Hybridization chain reaction circuit; MicroRNA detection; Signal amplification.

MeSH terms

Biosensing Techniques*
CRISPR-Cas Systems / genetics
DNA
MicroRNAs* / analysis
MicroRNAs* / genetics
Nucleic Acid Hybridization

Substances

MicroRNAs
DNA