Multiplexed Nanoflares: mRNA Detection in Live Cells

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• 作者：Andrew E. Prigodich ; Pratik S. Randeria ; William E. Briley ; Nathaniel J. Kim ; Weston L. Daniel ; David A. Giljohann ; Chad A. Mirkin
• 刊名：Analytical Chemistry
• 出版年：2012
• 出版时间：February 21, 2012
• 年：2012
• 卷：84
• 期：4
• 页码：2062-2066
• 全文大小：332K
• 年卷期：v.84,no.4(February 21, 2012)
• ISSN：1520-6882

文摘

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.