Ultrasensitive electrochemical assay for microRNA-21 based on CRISPR/Cas13a-assisted catalytic hairpin assembly

Ying Cui; Shanji Fan; Ze Yuan; Minghui Song; Jiawen Hu; Dong Qian; Deshuai Zhen; Junhua Li; Baode Zhu

doi:10.1016/j.talanta.2020.121878

Ultrasensitive electrochemical assay for microRNA-21 based on CRISPR/Cas13a-assisted catalytic hairpin assembly

Talanta. 2021 Mar 1:224:121878. doi: 10.1016/j.talanta.2020.121878. Epub 2020 Nov 9.

Authors

Ying Cui¹, Shanji Fan², Ze Yuan³, Minghui Song³, Jiawen Hu³, Dong Qian⁴, Deshuai Zhen⁵, Junhua Li⁶, Baode Zhu⁷

Affiliations

¹ Hunan Key Laboratory of Two-Dimensional Materials, Advanced Catalytic Engineering Research Center of the Ministry of Education, and College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082, China; College of Chemistry and Materials Science, Hengyang Normal University, Hengyang, 421001, China.
² Department of Thyroid Breast Surgery, The First Affiliated Hospital of University of South China, Hengyang, 421001, China.
³ Hunan Key Laboratory of Two-Dimensional Materials, Advanced Catalytic Engineering Research Center of the Ministry of Education, and College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082, China.
⁴ Hunan Provincial Key Laboratory of Chemical Power Resources, College of Chemistry and Chemical Engineering, Central South University, Changsha, 410083, PR China.
⁵ Hunan Key Laboratory of Two-Dimensional Materials, Advanced Catalytic Engineering Research Center of the Ministry of Education, and College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082, China. Electronic address: deshuaizhen@hnu.edu.cn.
⁶ College of Chemistry and Materials Science, Hengyang Normal University, Hengyang, 421001, China. Electronic address: junhua325@126.com.
⁷ College of Chemistry Biology & and Environmental Engineering, Xiangnan University, Chenzhou, 423043, China. Electronic address: baodezhu@xnu.edu.cn.

PMID: 33379087
DOI: 10.1016/j.talanta.2020.121878

Abstract

MicroRNAs (miRNAs) are related to many biological processes and regarded as biomarkers of disease. Rapid, sensitive, and specific methods for miRNA assay are very important for early disease diagnostic and therapy. In the present work, an ultrasensitive electrochemical biosensing platform has been developed for miRNA-21 assay by combining CRISPR-Cas13a system and catalytic hairpin assembly (CHA). In the presence of miRNA-21, it would hybridize with the spacer region of Cas13a/crRNA duplex to activate the cleavage activity of CRISPR-Cas13a system, leading to the release of initiator of CHA to generate amplified electrochemical signals. Base on the CRISPR-Cas13a-mediated cascade signal amplification strategy, the developed electrochemical biosensing platform exhibited high sensitivity with a low detection limit of 2.6 fM (S/N = 3), indicating that the platform has great potential for application in early clinical diagnostic.

Keywords: CRISPR-Cas13a system; Cascade signal amplification; Catalytic hairpin assembly; Electrochemical biosensor; miRNA-21.

MeSH terms

Biological Assay
Biosensing Techniques*
Catalysis
Clustered Regularly Interspaced Short Palindromic Repeats
Electrochemical Techniques
Limit of Detection
MicroRNAs* / genetics

Substances

MicroRNAs