期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 182, 期 -, 页码 156-160出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2013.02.113
关键词
Fluorescence; Graphene oxide; Argonaute2 protein; RNA endonuclease activity
资金
- National Natural Science Foundation of China [21175076, 21205065]
- Basic Scientific Fund for National Public Research Institutes of China [2012T07]
- Scientific Special Expenditure for Non-profit Public Industry of State Oceanic Administration [201105020]
- Basic Research Program of Qingdao [11-2-4-3-(4)-jch]
A novel graphene oxide (GO)-based amplified fluorescence assay for RNA endonuclease activity of mammalian Argonaute2 (Ago2) was developed. Owing to the extraordinarily high quenching efficiency of GO, the probe labeled with a fluorophore carboxyfluorescein exhibits minimal background fluorescence, while strong fluorescence intensity is observed with the addition of Ago2-miRNA complex due to the scission of the target mRNA sequences and the release of FAM-linked oligonucleotide fragments. Furthermore, Ago2-miRNA complex may be released to initiate another cycle of cleavage. The newly designed simple and cost-effective method exhibits high sensitivity with a detection limit of 1.7 nM (3 sigma) for Ago2 endonuclease activity. (C) 2013 Elsevier B.V. All rights reserved.
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