2. Academic Validation
  3. CRISPR-Cas12a-based efficient electrochemiluminescence biosensor for ATP detection

CRISPR-Cas12a-based efficient electrochemiluminescence biosensor for ATP detection

  • Anal Chim Acta. 2021 Dec 15:1188:339180. doi: 10.1016/j.aca.2021.339180.
Zhi-Hong Xu 1,  Zi-Yuan Zhao 1,  Hui Wang 1,  Shu-Min Wang 1,  Hong-Yuan Chen 1,  Jing-Juan Xu 2
Affiliations

Affiliations

  • 1 State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, China.
  • 2 State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, China. Electronic address: [email protected].
PMID: 34794559 DOI: 10.1016/j.aca.2021.339180
Abstract

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.

Keywords

Adenosine triphosphate; Bipolar electrode; CRISPR-Cas12a; Electrochemiluminescence.

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