Issue 24, 2022

Chemiluminescence resonance energy transfer as a simple and sensitive readout mode for a CRISPR/Cas12a-based biosensing platform

Abstract

Various CRISPR/Cas12a-based biosensing systems have been developed in the past few years, and most of these systems used Taqman probes to report signals through fluorescence resonance energy transfer (FRET). In this study, we explored chemiluminescence resonance energy transfer (CRET) as the readout mode for CRISPR/Cas12a-based biosensing. The chemiluminescence (CL) reaction of bis(2,4,6-trichlorophenyl) oxalate (TCPO) and H2O2 was used to excite the fluorophore dye of the Taqman probe. Different from FRET, CRET does not need external excitation light, which can effectively avoid autofluorescence and photobleaching. The detection limit of this CRET readout mode was estimated to be 10 pM for target DNA, which was about 8 times lower than that of the widely used FRET readout mode. These results suggest that CRET can serve as a rapid, sensitive and simple readout mode of CRISPR/Cas12-based biosensing, and can further enrich the toolbox of CRISPR/Cas12-based biosensing.

Graphical abstract: Chemiluminescence resonance energy transfer as a simple and sensitive readout mode for a CRISPR/Cas12a-based biosensing platform

Supplementary files

Article information

Article type
Paper
Submitted
22 Okt 2022
Accepted
13 Nov 2022
First published
16 Nov 2022

Analyst, 2022,147, 5687-5693

Chemiluminescence resonance energy transfer as a simple and sensitive readout mode for a CRISPR/Cas12a-based biosensing platform

Y. Zhang, J. Lei, W. Liu, Y. Jin and B. Li, Analyst, 2022, 147, 5687 DOI: 10.1039/D2AN01735K

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