Simple and cheap CRISPR/Cas12a biosensor based on plug-and-play of DNA aptamers for the detection of endocrine-disrupting compounds

Yunqiu Zhao; Lin Zhu; Yaxue Ding; Weiting Ji; Kun Liu; Ke Liu; Bei Gao; Xinyi Tao; Yu-Guo Dong; Feng-Qing Wang; Dongzhi Wei

doi:10.1016/j.talanta.2023.124761

Simple and cheap CRISPR/Cas12a biosensor based on plug-and-play of DNA aptamers for the detection of endocrine-disrupting compounds

Talanta. 2023 Oct 1:263:124761. doi: 10.1016/j.talanta.2023.124761. Epub 2023 May 30.

Authors

Yunqiu Zhao¹, Lin Zhu², Yaxue Ding², Weiting Ji², Kun Liu², Ke Liu¹, Bei Gao¹, Xinyi Tao¹, Yu-Guo Dong³, Feng-Qing Wang⁴, Dongzhi Wei¹

Affiliations

¹ State Key Laboratory of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China; Key Laboratory of Biocatalysis and Intelligent Manufacturing (ECUST), China National Light Industry, Shanghai, 200237, China.
² State Key Laboratory of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China.
³ Key Laboratory of Biocatalysis and Intelligent Manufacturing (ECUST), China National Light Industry, Shanghai, 200237, China. Electronic address: dongyuguo@ecust.edu.cn.
⁴ State Key Laboratory of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China; Key Laboratory of Biocatalysis and Intelligent Manufacturing (ECUST), China National Light Industry, Shanghai, 200237, China. Electronic address: fqwang@ecust.edu.cn.

PMID: 37267883
DOI: 10.1016/j.talanta.2023.124761

Abstract

Endocrine-disrupting compounds (EDCs) are widely distributed in the environment. Here, we present a CRISPR/Cas12a (CAS) biosensor based on DNA aptamers for point-of-care detection of EDCs. Two typical EDCs, 17β-estradiol (E2) and bisphenol A (BPA), were selected to be detected by the CAS biosensors via the plug-and-play of their DNA aptamers. The results indicated that the performance of the CAS biosensors can be well regulated by controlling the trans-cleavage activity of Cas12a on a single-stranded DNA reporter and optimizing the sequence and ratio of DNA aptamer and activator DNA. Ultimately, two reliable and specific biosensors were developed, with the linear range and limit of detection of 0.2-25 nM and 0.08 nM for E2 and of 0.1-250 nM and 0.06 nM for BPA, respectively. Compared to the existing detection methods, the CAS biosensors showed higher reliability and sensitivity with simple operation, short detection time, and no costly equipment.

Keywords: 17β-estradiol; Bisphenol A; CRISPR/Cas12a biosensor; DNA aptamers; Endocrine-disrupting compounds.

MeSH terms

Aptamers, Nucleotide* / genetics
Biosensing Techniques* / methods
CRISPR-Cas Systems
Estradiol
Reproducibility of Results

Substances

Aptamers, Nucleotide
Estradiol