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Issue 30, 2015
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Molecular beacon-loaded polymeric nanoparticles for non-invasive imaging of mRNA expression

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Abstract

Assessment of intracellular mRNA expression is invaluable for understanding cellular signaling activities, identifying disease stages, and monitoring the gene expression pattern of therapeutic cells during their culture, expansion and/or differentiation process. Previous methods suffer from the need to disrupt the biological samples to perform polymerase chain reaction analysis which can be laborious, fragmented and destructive. Herein, we develop a mRNA nanosensor based on the sustained release of mRNA-specific molecular beacons (probes that fluoresce upon hybridization) from the biodegradable poly(D,L-lactide-co-glycolide) nanoparticles. Post cellular internalization, the particles gradually degrade and release the encapsulated probes which are initially weakly fluorescent. When the released probes meet and hybridize with target mRNA, they restore pre-quenched fluorescence. By virtue of quantifying the fluorescence intensity, we can estimate the cellular mRNA expression. As a case study, β-actin mRNA expression in mesenchymal stem cells cultured on a 3D matrix was monitored and compared with those cultured on a 2D plate for one week. Critically, the observed expression profile shows a great correlation with the established quantitative polymerase chain reaction analysis.

Graphical abstract: Molecular beacon-loaded polymeric nanoparticles for non-invasive imaging of mRNA expression

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Publication details

The article was received on 10 May 2015, accepted on 30 Jun 2015 and first published on 01 Jul 2015


Article type: Communication
DOI: 10.1039/C5TB00876J
Citation: J. Mater. Chem. B, 2015,3, 6148-6156
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    Molecular beacon-loaded polymeric nanoparticles for non-invasive imaging of mRNA expression

    C. Wiraja, D. C. Yeo, S. Y. Chew and C. Xu, J. Mater. Chem. B, 2015, 3, 6148
    DOI: 10.1039/C5TB00876J

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