title: Multiplexed nanoflares: mRNA detection in live cells.
abstract: 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.
authors.affiliation: Department of Chemistry and International Institute for Nanotechnology, Northwestern University, 2145 Sheridan Road, Evanston, Illinois 60208-3113, USA.
authors.forename: Andrew E
authors.initials: AE
authors.lastname: Prigodich
authors.forename: Pratik S
authors.initials: PS
authors.lastname: Randeria
authors.forename: William E
authors.initials: WE
authors.lastname: Briley
authors.forename: Nathaniel J
authors.initials: NJ
authors.lastname: Kim
authors.forename: Weston L
authors.initials: WL
authors.lastname: Daniel
authors.forename: David A
authors.initials: DA
authors.lastname: Giljohann
authors.forename: Chad A
authors.initials: CA
authors.lastname: Mirkin
mesh_terms: D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006046:Gold / Q000737:chemistry*; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics*; D053768:Metal Nanoparticles / Q000737:chemistry*; D053758:Nanoparticles; D009369:Neoplasms / Q000235:genetics; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077022:Survivin; D014407:Tumor Cells, Cultured
chemical_list: C107489:BIRC5 protein, human; D005456:Fluorescent Dyes; D051034:Inhibitor of Apoptosis Proteins; D012333:RNA, Messenger; D000077022:Survivin; D006046:Gold