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 Cas12a-crRNA 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. Sensing mechanism for ATP detection. [Display omitted] • The paper provides an efficient ECL biosensing system based on CRISPR-Cas12a for the determination of ATP. • CRISPR-Cas12a can convert the recognition of target ATP into detectable ECL signals on bipolar electrodes. • Sensitive quantitative analysis of ATP was achieved with the detection limit of 0.48 nM. [ABSTRACT FROM AUTHOR]