title: Quantification and real-time tracking of RNA in live cells using Sticky-flares.
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 Sticky-flares 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.
authors.affiliation: Department of Interdisciplinary Biological Sciences, Northwestern University, Evanston, IL 60208; International Institute for Nanotechnology, Northwestern University, Evanston, IL 60208;
authors.forename: William E
authors.initials: WE
authors.lastname: Briley
authors.affiliation: Department of Chemistry, Northwestern University, Evanston, IL 60208;
authors.forename: Madison H
authors.initials: MH
authors.lastname: Bondy
authors.affiliation: International Institute for Nanotechnology, Northwestern University, Evanston, IL 60208; Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208.
authors.forename: Pratik S
authors.initials: PS
authors.lastname: Randeria
authors.affiliation: Department of Chemistry, Northwestern University, Evanston, IL 60208;
authors.forename: Torin J
authors.initials: TJ
authors.lastname: Dupper
authors.affiliation: Department of Interdisciplinary Biological Sciences, Northwestern University, Evanston, IL 60208; International Institute for Nanotechnology, Northwestern University, Evanston, IL 60208; Department of Chemistry, Northwestern University, Evanston, IL 60208; Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208 chadnano@northwestern.edu.
authors.forename: Chad A
authors.initials: CA
authors.lastname: Mirkin
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D002470:Cell Survival; D058948:Cell Tracking / Q000379:methods*; D003199:Computer Systems; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006046:Gold / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D053768:Metal Nanoparticles / Q000737:chemistry; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D012313:RNA / Q000378:metabolism*; D034443:RNA Transport; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D012333:RNA, Messenger; D012313:RNA; D006046:Gold
keywords: FISH; RNA detection; RNA localization; in situ hybridization; nanotechnology