Visual detection of Akt mRNA in living cell using gold nanoparticle beacon

Proceedings of SPIE(2014)

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摘要
PI3K-Akt signaling pathway plays the key role in cell apoptosis and survival, and the components of PI3K /Akt signaling pathway are often abnormally expressed in human tumors. Therefore, determination of the Akt (protein kinase B, PKB) messenger ribonucleic acid (mRNA) expression is significantly important in understanding the mechanism of tumor progression. In this study, we designed a special hairpin deoxyribonucleic acid (DNA) functionalized with gold nanoparticles and fluorescein isothiocyanate(FITC) as a beacon for detecting human Akt mRNA. Spectrofluorometer was used to detect the fluorescence quenching and recovery of the beacons, and laser confocal scanning microscopy was adopted to image Akt mRNA in cells. The results showed that this beacon could sensitively and quantitatively measure the Akt mRNA in living cells. This strategy is potentially useful for the cellular imaging of RNA or protein expression in living cells.
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关键词
fluorescence imaging,PI3K-Akt,molecular beacon,gold nanoparticles,living cell,fluorescent dyes
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