Journal
JOURNAL OF SOLID STATE ELECTROCHEMISTRY
Volume 18, Issue 10, Pages 2829-2835Publisher
SPRINGER
DOI: 10.1007/s10008-014-2531-y
Keywords
Electrochemical biosensor; miRNA-159a; Label-free detection; Cu-apoferritin
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Funding
- National Natural Science Foundation of China [21075078, 21105056]
- Natural Science Foundation of Shandong province, China [ZR2010BM005, ZR2011BQ001]
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MicroRNAs (miRNAs), a class of small endogenous nonprotein-coding RNAs, regulate a wide range of biological processes, and their abnormal expressions are related to the growth and development of plants. Thus, a simple, rapid, and highly sensitive assay for miRNA detection is of great significance. In this work, a label-free and ultrasensitive assay for miRNA detection using protein cage nanoparticles has been developed. Apoferritin-encapsulated Cu nanoparticles (Cu-apoferritin) could be immobilized on the electrode through special reaction between amino and carboxyl. Hybridization event between the probe DNA and the target miRNA-159a is confirmed by electrochemical oxidation signal after Cu released into the detection buffer by adjusting the pH. This assay is highly selective and sensitive with a low detection limit of 3.5 fM. Moreover, the developed method can even discriminate single-base mismatched strand between the complementary targets. The effect of abscisic acid on the expression level of miRNA-159a in Arabidopsis thaliana seeds was also investigated.
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