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Issue 17, 2013
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Detection of mRNA in living cells by double-stranded locked nucleic acid probes

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Abstract

Double-stranded probes are homogeneous biosensors for rapid detection of specific nucleotide sequences. These double-stranded probes have been applied in various molecular sensing applications, such as real-time polymerase chain reaction and detection of bacterial 16S rRNA. In this study, we present the design and optimization of double-stranded probes for single-cell gene expression analysis in living cells. With alternating DNA/LNA monomers for optimizing the stability and specificity, we show that the probe is stable in living cells for over 72 hours post-transfection and is capable of detecting changes in gene expression induced by external stimuli. The probes can be delivered to a large number of cells simultaneously by cationic liposomal transfection or to individual cells selectively by photothermal delivery. We also demonstrate that the probe quantifies intracellular mRNA in living cells through the use of an equilibrium analysis. With its effectiveness and performance, the double-stranded probe represents a broadly applicable approach for large-scale single-cell gene expression analysis toward numerous biomedical applications, such as systems biology, cancer, and drug screening.

Graphical abstract: Detection of mRNA in living cells by double-stranded locked nucleic acid probes

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Publication details

The article was received on 11 Apr 2013, accepted on 16 May 2013 and first published on 17 Jun 2013


Article type: Paper
DOI: 10.1039/C3AN00722G
Citation: Analyst, 2013,138, 4777-4785
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    Detection of mRNA in living cells by double-stranded locked nucleic acid probes

    R. Riahi, Z. Dean, T. Wu, M. A. Teitell, P. Chiou, D. D. Zhang and P. K. Wong, Analyst, 2013, 138, 4777
    DOI: 10.1039/C3AN00722G

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