Catalytic Hairpin Assembly-Aided CRISPR/Cas-SERS for Nucleic Acid Detection Enhanced by Multiple Catalytic Amplification.
5 affiliations
- 1State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun 130012, P. R. China.
- 2School of Chemistry and Environmental Engineering, Changchun University of Science and Technology, Changchun 130022, P. R. China.
- 3Institute of Agricultural Quality Standard and Testing Technology, Jilin Academy of Agricultural Sciences (Northeast Innovation Center of Agricultural Science and Technology in China), Changchun 130033, P. R. China.
- 4Maize Research Institute, Jilin Academy of Agricultural Sciences (Northeast Innovation Center of Agricultural Science and Technology in China), Changchun 130118, P.R. China.
- 5Center for Supramolecular Chemical Biology, College of Chemistry, Jilin University, Changchun 130012, P. R. China.
Abstract
This study established a surface-enhanced Raman scattering (SERS) detection method based on the catalytic hairpin assembly (CHA) reaction combined with clustered regularly interspaced short palindromic repeats and its associated proteins (CRISPR/Cas), integrated with the guanine-quadruplexes/Hemin (G4/Hemin) catalytic amplification effect, which is available for identifying a typical indicator for genetically modified crops, the 35S promoter from the cauliflower mosaic virus (CaMV35S). Target CaMV35S-derived RNA (CaMV35S RNA) triggers the CHA reaction, yielding a double-stranded DNA product (C1/C2) that is recognized by CRISPR RNA (crRNA), thereby activating the trans-cleavage activity of Cas12a. A magnetic bead-based nanoprobe was fabricated by linking the hybridization chain reaction (HCR) product to the magnetic bead. This probe can fold into repeated G4 units under a specific potassium ion concentration to enable a high loading of Hemin, thereby providing strong peroxidase-like catalytic activity for 3,3',5,5'-tetra-methyl-benz-idine (TMB). In a positive trial, the 35S RNA initiates CHA, and the CHA product activates Cas12a, disrupting the HCR products on the magnetic bead probe, preventing the G4 structure formation, accordingly yielding a weak SERS signal from TMB. In contrast, a negative trial fails to activate Cas12a, and the probe retains its nanoenzyme activity, resulting in a strong SERS signal from the produced oxTMB. Owing to the collateral trans-cleavage activity of Cas12a and the high peroxidase-like nanozyme activity of G4/Hemin, this method achieves the highly sensitive SERS detection of genetically modified samples without target gene preamplification.
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BibTeX
@article{liu2026catalytic,
title = {{Catalytic Hairpin Assembly-Aided CRISPR/Cas-SERS for Nucleic Acid Detection Enhanced by Multiple Catalytic Amplification.}},
author = {Lin Liu and Chengxin Bao and Huimin Wang and Feiwu Li and Xiangguo Liu and Yuxuan He and Jingjing Chang and Yuejia Yin and Shuping Xu},
journal = {ACS measurement science au},
year = {2026},
month = aug,
doi = {10.1021/acsmeasuresciau.6c00142},
url = {https://europepmc.org/article/MED/42921428},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Catalytic Hairpin Assembly-Aided CRISPR/Cas-SERS for Nucleic Acid Detection Enhanced by Multiple Catalytic Amplification.
AU - Lin Liu
AU - Chengxin Bao
AU - Huimin Wang
AU - Feiwu Li
AU - Xiangguo Liu
AU - Yuxuan He
AU - Jingjing Chang
AU - Yuejia Yin
AU - Shuping Xu
PY - 2026
DA - 2026/08/21
JO - ACS measurement science au
DO - 10.1021/acsmeasuresciau.6c00142
UR - https://europepmc.org/article/MED/42921428
AB - This study established a surface-enhanced Raman scattering (SERS) detection method based on the catalytic hairpin assembly (CHA) reaction combined with clustered regularly interspaced short palindromic repeats and its associated proteins (CRISPR/Cas), integrated with the guanine-quadruplexes/Hemin (G4/Hemin) catalytic amplification effect, which is available for identifying a typical indicator for genetically modified crops, the 35S promoter from the cauliflower mosaic virus (CaMV35S). Target CaMV35S-derived RNA (CaMV35S RNA) triggers the CHA reaction, yielding a double-stranded DNA product (C1/C2) that is recognized by CRISPR RNA (crRNA), thereby activating the trans-cleavage activity of Cas12a. A magnetic bead-based nanoprobe was fabricated by linking the hybridization chain reaction (HCR) product to the magnetic bead. This probe can fold into repeated G4 units under a specific potassium ion concentration to enable a high loading of Hemin, thereby providing strong peroxidase-like catalytic activity for 3,3',5,5'-tetra-methyl-benz-idine (TMB). In a positive trial, the 35S RNA initiates CHA, and the CHA product activates Cas12a, disrupting the HCR products on the magnetic bead probe, preventing the G4 structure formation, accordingly yielding a weak SERS signal from TMB. In contrast, a negative trial fails to activate Cas12a, and the probe retains its nanoenzyme activity, resulting in a strong SERS signal from the produced oxTMB. Owing to the collateral trans-cleavage activity of Cas12a and the high peroxidase-like nanozyme activity of G4/Hemin, this method achieves the highly sensitive SERS detection of genetically modified samples without target gene preamplification.
SN - 2694-250X
ER -