Multiplexed Nanoflares: mRNA Detection in Live Cells

We report the development of the multiplexed nanoflare, a nanoparticle agent that is capable of simultaneously detecting two distinct mRNA targets inside a living cell. These probes are spherical nucleic acid (SNA) gold nanoparticle (Au NP) conjugates consisting of densely packed and highly oriented...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2012-02, Vol.84 (4), p.2062-2066
Main Authors: Prigodich, Andrew E, Randeria, Pratik S, Briley, William E, Kim, Nathaniel J, Daniel, Weston L, Giljohann, David A, Mirkin, Chad A
Format: Article
Language:English
Subjects:
Analytical chemistry
Applied sciences
Cells
Chemistry
Exact sciences and technology
Flow Cytometry
Fluorescent Dyes
Global environmental pollution
Gold - chemistry
Humans
Inhibitor of Apoptosis Proteins - genetics
Metal Nanoparticles - chemistry
Nanoparticles
Neoplasms - genetics
Nucleic Acid Hybridization
Pollution
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
Ribonucleic acid
RNA
RNA, Messenger - analysis
RNA, Messenger - genetics
Tumor Cells, Cultured
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Summary:We report the development of the multiplexed nanoflare, a nanoparticle agent that is capable of simultaneously detecting two distinct mRNA targets inside a living cell. These probes are spherical nucleic acid (SNA) gold nanoparticle (Au NP) conjugates consisting of densely packed and highly oriented oligonucleotide sequences, many of which are hybridized to a reporter with a distinct fluorophore label and each complementary to its corresponding mRNA target. When multiplexed nanoflares are exposed to their targets, they provide a sequence specific signal in both extra- and intracellular environments. Importantly, one of the targets can be used as an internal control, improving detection by accounting for cell-to-cell variations in nanoparticle uptake and background. Compared to single-component nanoflares, these structures allow one to determine more precisely relative mRNA levels in individual cells, improving cell sorting and quantification.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac202648w