期刊
BIOSENSORS & BIOELECTRONICS
卷 60, 期 -, 页码 351-357出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2014.04.040
关键词
Poly-cytosine nucleotide loop; Hybridization chain reaction; Fluorescent silver nanoclusters; Single nucleotide polymorphisms; Let-7 miRNA family
类别
资金
- National Natural Science Foundation of China [21175027, 20975026]
- Shanghai Natural Science Foundation of China [12ZR1401800]
A simple microRNA (miRNA) detection system based on hybridization chain reaction (HCR) has been developed using highly fluorescent DNA-hosted silver (Ag) nanoclusters. In this assay, a new type of hairpin DNA probe (MB1) containing a poly-cytosine nucleotide loop is designed and used as one of the HCR monomers, which is also demonstrated to be an ideal template for in situ synthesis of highly fluorescent Ag nanoclusters. Correspondingly, another HCR monomer (MB2) contains a poly-guanine nucleotide sticky end. Two monomers are stable to coexist in solution until the introduction of the initiator strand (let-7a) triggers a cascade of hybridization events that yields nicked double helices analogous. By taking advantage of HCR, a small amount of let-7a leads to the conformational change of a large amount of MB1, which results in the decrease of fluorescent signal greatly. Overall, this label-free, enzyme-free method allows the sensitive detection of let-7a with high specificity towards single nucleotide polymorphisms in the let-7 miRNA family. In addition, the simple mix and measure assay can be extended to detect other types of targets upon slight modification, and thus provides a tool for the early diagnosis and risk assessment of malignancy. (C) 2014 Elsevier B.V. All rights reserved.
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