Quantification and real-time tracking of RNA in live cells using Sticky-flares

William E. Briley, Madison H. Bondy, Pratik S. Randeria, Torin J. Dupper, Chad A. Mirkin*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

57 Scopus citations

Abstract

We report a novel spherical nucleic acid (SNA) gold nanoparticle conjugate, termed the Sticky-flare, which enables facile quantification of RNA expression in live cells and spatiotemporal analysis of RNA transport and localization. The Sticky-flare is capable of entering live cells without the need for transfection agents and recognizing target RNA transcripts in a sequence-specific manner. On recognition, the Sticky-flare transfers a fluorophore-conjugated reporter to the transcript, resulting in a turning on of fluorescence in a quantifiable manner and the fluorescent labeling of targeted transcripts. The latter allows the RNA to be tracked via fluorescence microscopy as it is transported throughout the cell. We use this novel nanoconjugate to analyze the expression level and spatial distribution of β-actin mRNA in HeLa cells and to observe the real-time transport of β-actin mRNA in mouse embryonic fibroblasts. Furthermore, we investigate the application of Stickyflares for tracking transcripts that undergo more extensive compartmentalization by fluorophore-labeling U1 small nuclear RNA and observing its distribution in the nucleus of live cells.

Original languageEnglish (US)
Pages (from-to)9591-9595
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume112
Issue number31
DOIs
StatePublished - Aug 4 2015

Keywords

  • FISH
  • In situ hybridization
  • Nanotechnology
  • RNA detection
  • RNA localization

ASJC Scopus subject areas

  • General

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