CRISPR/Cas12a-Assisted Visual Logic-Gate Detection of Pathogenic Microorganisms Based on Water-Soluble DNA-Binding AIEgens

Zhe Jiao; Jialing Yang; Xiaojuan Long; Yingfang Lu; Zongning Guo; Yonglin Peng; Xuelin Huang; Yi Yin; Chao Song; Pengfei Zhang

doi:10.3389/fchem.2021.801972

CRISPR/Cas12a-Assisted Visual Logic-Gate Detection of Pathogenic Microorganisms Based on Water-Soluble DNA-Binding AIEgens

Front Chem. 2022 Jan 14:9:801972. doi: 10.3389/fchem.2021.801972. eCollection 2021.

Authors

Zhe Jiao¹, Jialing Yang¹, Xiaojuan Long², Yingfang Lu², Zongning Guo³, Yonglin Peng⁴, Xuelin Huang³, Yi Yin⁵, Chao Song^{6

7}, Pengfei Zhang⁸

Affiliations

¹ School of Environment and Civil Engineering, Dongguan University of Technology, Dongguan, China.
² Guangdong Dongguan Ecological and Environmental Monitoring Station, Dongguan, China.
³ Huangpu Customs District Technology Center, Dongguan, China.
⁴ Pinete (Zhongshan) Biotechnology Co., Ltd., Zhongshan, China.
⁵ Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China.
⁶ Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, China.
⁷ Key Laboratory of Control of Quality and Safety for Aquatic Products, Ministry of Agriculture and Rural Affairs, Beijing, China.
⁸ Guangdong Key Laboratory of Nanomedicine, Shenzhen Engineering Laboratory of Nanomedicine and Nanoformulations, CAS Key Laboratory of Health Informatics, Institute of Biomedicine and Biotechnology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.

Abstract

Here, we developed a rapid, visual and double-checked Logic Gate detection platform for detection of pathogenic microorganisms by aggregation-induced emission luminogens (AIEgens) in combination with Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR associated (Cas). DNA light-up AIEgens (1,1,2,2-tetrakis[4-(2-bromo-ethoxy) phenyl]ethene, TTAPE) was non-emissive but the emission was turned on in the presence of large amount of DNA produced by recombinase polymerase amplification (RPA). When CRISPR/Cas12a was added, all long-stranded DNA were cut leading to the emission quenched. Thus, a method that can directly observe the emission changes with the naked eye has been successfully constructed. The detection is speedy within only 20 min, and has strong specificity to the target. The result can be judged by Logic Gate. Only when the output signal is (1,0), does it represent the presence of pathogenic microorganisms in the test object. Finally, the method was applied to the detect pathogenic microorganisms in environmental water samples, which proved that this method has high selectivity, specificity and applicability for the detection of pathogenic microorganisms in environmental water samples.

Keywords: CRISPR-Cas12a; aggregation-induced emission (AIE); logic gate; pathogenic microorganisms; visual detection.