期刊
BIOSENSORS & BIOELECTRONICS
卷 216, 期 -, 页码 -出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2022.114646
关键词
CRISPR; Biosensor; Imaging; Live cell; In vivo
类别
资金
- National Natural Science Foundation of China [21904102, 81802053]
- Research Foundation for Talented Scholars of Hubei University of Medicine [2020QDJZR001]
In this study, a CRISPR/Cas12a-based biosensor is proposed that achieves highly-efficient biological uptake through the physical absorption of multiple biomolecular components by MnO(2) nanosheets. A smart near-infrared (NIR) light-gated manner is designed to offer spatiotemporal target recognition, preventing pre-initiation during biological delivery. The biosensor demonstrates satisfactory sensitivity and specificity to the cancer biomarker survivn messenger RNA and can be used for reliable imaging and analysis in live cellular and mouse models, greatly advancing CRISPR technology in the field of biosensing.
Despite the in vitro usage of CRISPR/Cas12a system in fluorescent biosensors has made remarkable achievements, many challenges such as poor biological delivery, insufficient sensitivity, and uncontrollable initiation compel them hard to conduct in vivo analysis. Thus, we propose here some fruitful sensing concepts. First, the multiple biomolecular components of CRISPR/Cas12a system are collectively carried by MnO(2 )nanosheets via a simple physical absorption to achieve a highly-efficient biological uptake. Under the reduction of widespread biothiols, not only the sensing frame is easily released but also sufficient Mn2+ is produced to serve as an effective trans-cleavage accelerator. Furthermore, a photocleavge-linker induced smart near-infrared (NIR) light-gated manner is designed to offer a spatiotemporal target recognition, for which a 808 nm NIR light transduced ul-traviolet upconversion luminescence with weak thermal effect is employed to completely prevent the sensing flow from pre-initiating during the biological delivery. As a conceptual validation, this biosensor has satisfactory sensitivity and specificity to survivn messenger RNA (a broad-spectrum cancer biomarker). More importantly, it can work as a reliable imaging platform for differentiating cancers in live cellular level and also presents a high-performance operation ability for analyzing live mice, greatly promoting the CRISPR technology in biosensing field.
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