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

Content Provider	PubMed Central
Author	Riahi, Reza Dean, Zachary Wu, Ting-hsiang Teitell, Michael A. Chiou, Pei-yu Zhang, Donna D. Wong, Pak Kin
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.
Related Links	http://dx.doi.org/10.1039/c3an00722g
Ending Page	4785
Page Count	9
Starting Page	4777
File Format	PDF
ISSN	00032654
e-ISSN	13645528
Journal	The Analyst
Issue Number	17
Volume Number	138
Language	English
Publisher Date	2013-01-01
Access Restriction	Open
Subject Keyword	Analytical Chemistry Spectroscopy Electrochemistry Biochemistry Environmental Chemistry Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Spectroscopy Environmental Chemistry Analytical Chemistry Biochemistry Electrochemistry

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