Label-Free and Universal CRISPR/Cas12a-Based Detection Platform for Nucleic Acid Biomarkers

CRISPR/Cas12a has been widely used in molecular diagnostics due to its excellent trans-cleavage activity. However, conventional reporters, such as F/Q-labeled single-stranded DNA (ssDNA) reporters, enzyme-labeled reporters, and spherical nucleic acid reporters, require complex modification or labeli...

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Published in:ACS sensors 2024-09, Vol.9 (9), p.4803-4810
Main Authors: Zhao, Rujian, Xiao, Yao, Tang, Yidan, Lu, Baiyang, Li, Bingling
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creator Zhao, Rujian
Xiao, Yao
Tang, Yidan
Lu, Baiyang
Li, Bingling
description CRISPR/Cas12a has been widely used in molecular diagnostics due to its excellent trans-cleavage activity. However, conventional reporters, such as F/Q-labeled single-stranded DNA (ssDNA) reporters, enzyme-labeled reporters, and spherical nucleic acid reporters, require complex modification or labeling processes. In this study, we have developed a rapid, universal, and label-free CRISPR/Cas12a-based biomarker detection platform via designing a G-quadruplex (G4) containing a hairpin structure as the reporter. The hairpin loop design of hairpin G4 improves the cleavage efficiency of Cas12a and the signal strength of the G4 binding ligand. Meanwhile, the incorporation of a G4 binding dye (protoporphyrin IX) eliminates the need for complex modifications. The CRISPR-hairpin G4 detection platform is capable of detecting ssDNA, double-stranded DNA, genetic RNAs, and miRNAs. Moreover, this platform achieves label-free detection in clinical samples, demonstrating its practical applicability and efficiency.
doi_str_mv 10.1021/acssensors.4c01233
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title Label-Free and Universal CRISPR/Cas12a-Based Detection Platform for Nucleic Acid Biomarkers
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