期刊
NANO LETTERS
卷 18, 期 8, 页码 5116-5123出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.nanolett.8b02004
关键词
Sensor; photoprotective group; photocleavable; modified electrode; caged hairpin; light-induced
类别
资金
- National Basic Research Program of China (973 Program) [2015CB932600]
- National Key R&D Program of China [2017YFA0208000, 2016YFF0100800]
- National Natural Science Foundation of China [21525523, 21722507, 21574048]
- Fok Ying-Tong Education Foundation, China [151011]
The spatiotemporal detection of a target mRNA in a single living cell is a major challenge in nanoscience and nanomedicine. We introduce a versatile method to detect mRNA at a single living cell level that uses photocleavable hairpin probes as functional units for the optical (fluorescent) and electrochemical (voltammetric) detection of MnSOD mRNA in single MCF-7 cancer cells. The fluorescent probe is composed of an ortho-nitrophenylphosphate ester functionalized hairpin that includes the FAM fluorophore in a caged configuration quenched by Dabcyl. The fluorescent probe is further modified with the AS 1411 aptamer to facilitate the targeting and internalization of the probe into the MCF-7 cells. Under UV irradiation, the hairpin is cleaved, leading to the intracellular mRNA toehold-stimulated displacement of the FAM-functionalized strand resulting in a switched-on fluorescence signal upon the detection of the mRNA in a single cell. In addition, a nanoelectrode functionalized with a methylene blue (MB) redox-active photocleavable hairpin is inserted into the cytoplasm of a single MCF-7 cell. Photocleavage of the hairpin leads to the mRNA-mediated toehold displacement of the redox-active strand associated with the probe, leading to the depletion of the voltammetric response of the probe. The parallel optical and electrochemical detection of the mRNA at a single cell level is demonstrated.
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