A CRISPR-Cas12a-mediated dual-mode luminescence and colorimetric nucleic acid biosensing platform based on upconversion nanozyme

Jiaxiang Yan; Bohan Yin; Qin Zhang; Chuanqi Li; Jiareng Chen; Yingying Huang; Jianhua Hao; Changqing Yi; Yu Zhang; Siu Hong Dexter Wong; Mo Yang

doi:10.1016/j.bios.2024.116963

A CRISPR-Cas12a-mediated dual-mode luminescence and colorimetric nucleic acid biosensing platform based on upconversion nanozyme

Biosens Bioelectron. 2024 Nov 19:270:116963. doi: 10.1016/j.bios.2024.116963. Online ahead of print.

Authors

Jiaxiang Yan¹, Bohan Yin², Qin Zhang¹, Chuanqi Li¹, Jiareng Chen¹, Yingying Huang¹, Jianhua Hao³, Changqing Yi⁴, Yu Zhang⁵, Siu Hong Dexter Wong⁶, Mo Yang⁷

Affiliations

¹ Department of Biomedical Engineering, The Hong Kong Polytechnic University, Hong Kong, 999077, China.
² Laboratory for Marine Drugs and Bioproducts, Qingdao Marine Science and Technology Center, Qingdao, 266237, China; School of Medicine and Pharmacy, Ocean University of China, Qingdao, 266003, China.
³ Department of Applied Physics, The Hong Kong Polytechnic University, Hong Kong, 999077, China.
⁴ Key Laboratory of Sensing Technology and Biomedical Instruments Guangdong, School of Biomedical Engineering, Sun Yat-Sen University, Guangzhou, 510006, China.
⁵ Department of Mechanical and Automotive Engineering, Royal Melbourne Institute of Technology, Melbourne, VIC, 3000, Australia.
⁶ Laboratory for Marine Drugs and Bioproducts, Qingdao Marine Science and Technology Center, Qingdao, 266237, China; School of Medicine and Pharmacy, Ocean University of China, Qingdao, 266003, China. Electronic address: dexterwong@ouc.edu.cn.
⁷ Department of Biomedical Engineering, The Hong Kong Polytechnic University, Hong Kong, 999077, China; Joint Research Center of Biosensing and Precision Theranostics, The Hong Kong Polytechnic University, Kowloon, Hong Kong, 999077, China. Electronic address: mo.yang@polyu.edu.hk.

PMID: 39603211
DOI: 10.1016/j.bios.2024.116963

Abstract

In this study, a CRISPR-Cas12a-mediated dual-mode upconversion luminescence/colorimetric nucleic acid biosensing platform is developed based on UCNP@SiO₂/CeO₂ (UNSC) nanozyme. Here, UNSC is conjugated with single-stranded DNA (ssDNA) probes used as both peroxidase-like nanozyme and upconversion luminescence donors. When no target nucleic acid is present, ssDNA-conjugated UNSC attaches on magnetic graphene oxide (MGO) via pi-pi stacking force, resulting in upconversion luminescence quenching (OFF) and no color change after magnetic removal of nanozymes attached on the MGO. In the presence of target nucleic acid, Cas12a is specifically activated by targeted nucleic acid and indiscriminately cleaves the ssDNA probes on UNSCs. UNSCs then detach from the MGO surface due to the weakening of binding force, leading to upconversion luminescence recovery (ON) and colorimetric change due to the existence of free nanozyme in the 3,3',5,5'-tetramethyl-benzidine assay. As a proof-of-concept, this biosensing platform shows a limit of detection of around 320 fM in the upconversion luminescence mode and ∼28.4 pM in the colorimetric mode for nucleic acid detection, respectively. This UNSC nanozyme-based CRISPR-Cas12a dual-mode biosensing system also demonstrates high selectivity, good repeatability, and facile operation, which allows easy adaption to other nucleic acid-based detection only by redesigning the sequence of CRISPR RNA.

Keywords: CRISPR-Cas12a; Dual-modality detection; Peroxidase-mimic nanozyme; Upconversion nanozyme.