CRISPR-Cas12a-based efficient electrochemiluminescence biosensor for ATP detection

CRISPR-Cas12a system exhibits tremendous potential in accurate recognition and quantitation of nucleic acids and non-nucleic-acid targets thanks to the discovery of its cleavage capability toward single-stranded DNA (ssDNA). In this study, we developed an efficient electrochemiluminescence (ECL) sensing platform based on CRISPR-Cas12a for the analysis of adenosine triphosphate (ATP). In the presence of the target, the successful release of the DNA activator is specially recognized by Cas12acrRNA duplex and activates the cleavage of ferrocene (Fc) labeled-ssDNA (Fc-ssDNA) modified on the cathode of bipolar electrode (BPE), resulting in a decrease of ECL intensity of [Ru(bpy)(3)](2+) /TPrA in the anodic cell of BPE. By means of the unique combination of Cas12a with ECL technique based on BPE, it can convert the recognition of target ATP into a detectable ECL signal. The detection limit of ATP was determined to be 0.48 nM under the optimal conditions. This work will expand the application of CRISPR-Cas detection system and propose a potential method for the analysis of non-nucleic-acid targets. (C) 2021 Elsevier B.V. All rights reserved.

Automated summary

The CRISPR-Cas12a system, with its cleavage capability toward ssDNA, shows great potential in accurate recognition and quantitation of nucleic acids and non-nucleic-acid targets. An efficient ECL sensing platform based on CRISPR-Cas12a was developed for ATP analysis, which converts the recognition of target ATP into a detectable ECL signal. The detection limit of ATP was determined to be 0.48 nM under the optimal conditions, expanding the application of the CRISPR-Cas detection system.