Peptide antisense nanoparticles

Patel, Pinal C.; Giljohann, David A.; Seferos, Dwight S.; Mirkin, Chad A.

doi:10.1073/pnas.0801609105

Cited by 103 publications

(81 citation statements)

References 59 publications

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“…Contrastingly, MEFs treated with Sticky-flares targeting U1 snRNA distinctly showed internuclear fluorescence. Importantly, SNA constructs are sequestered to the cytosol of live cells and cannot enter the nucleus on their own (33). Thus, the internuclear fluorescence of U1 Sticky-flares indicates specific release from the nanoparticle surface and subsequent transport of the fluorescentlabeled RNA target into the nucleus.…”

Section: Resultsmentioning

confidence: 99%

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

Briley

Bondy

Randeria

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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105

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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.he study of RNA is a critical component of biological research and in the diagnosis and treatment of disease. Recently, the localization of mRNA has been identified as an essential process for a number of cellular functions, including restricting the production of certain proteins to specific compartments within cells (1). For instance, synaptic potentiation, the basis of learning and memory, relies on the local translation of specific mRNAs in pre-and postsynaptic compartments (2). Likewise, the misregulation of RNA distribution is associated with many disorders, including mental retardation, autism, and cancer metastasis (3-5). However, despite the significant role of mRNA transport and localization in cellular function, the available methods to visualize these phenomena are severely limited. For example, FISH, the most commonly used technique to analyze spatial distribution of RNA, requires fixation and permeabilization of cells before analysis (6). As a result, analysis of dynamic RNA distribution is restricted to a single snapshot in time (7,8). With such a limitation, understanding the translocation of RNA with respect to time, cell cycle, or external stimulus is difficult if not impossible. Furthermore, fixed cell analysis is a lengthy and highly specialized procedure due to the number of steps necessary to prepare a sample. Fixation, permeabilization, blocking, and staining processes each require optimization and vary based on cell type and treatment conditions, rendering FISH prohibitively complicated in many cases. Likewise, live cell analysis platforms such as molecular beacons require toxic transfection techniques, such as microinjection or lipid transfection, and are rapidly sequestered to the nucleus on cellular entry (9, 10). Recently more sophisticated live cell analyses have been developed that use genetic engineering to introduce exogenous hybrid gene...

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Section: Resultsmentioning

confidence: 99%

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

Briley

Bondy

Randeria

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

105

View full text Add to dashboard Cite

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“…The details of this preparation are described elsewhere (Jeynes et al, 2013a), where we also show that the TAT coating compared to a FBS coating, increases the number of GNPs in the cell by about 5 times. Of note, it has been shown that coating the GNPs with bovine serum increases the diameter of the GNP by about 10nm (Tsai et al, 2011), while coating with TAT increases it by about 30nm (Patel et al, 2008).…”

Section: Gold Nanoparticle Treatmentmentioning

confidence: 99%

Investigation of gold nanoparticle radiosensitization mechanisms using a free radical scavenger and protons of different energies

et al. 2014

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Gold nanoparticles (GNPs) have been shown to sensitise cancer cells to X-ray radiation, particularly at kV energies where photoelectric interactions dominate and the high atomic number of gold makes a large difference to X-ray absorption. Protons have a high cross-section for gold at a large range of relevant clinical energies, and so potentially could be used with GNPs for increased therapeutic effect.Here, we investigate the contribution of secondary electron emission to cancer cell radiosensitisation and investigate how this parameter is affected by proton energy and a free radical scavenger. We simulate the emission from a realistic cell phantom containing GNPs after traversal by protons and X-rays with different energies. We find that with a range of proton energies (1 -250 MeV) there is a small increase in secondaries compared to a much larger increase with X-rays. Secondary electrons are known to produce toxic free radicals. Using a cancer cell line in vitro we find that a free radical scavenger has no protective effect on cells containing GNPs irradiated with 3 MeV protons, while it does protect against cells irradiated with X-rays.3

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“…Building on their previous work with proteins facilitating intracellular trafficking, the peptides bound to the nanoparticles were linked to increased cellular uptake and/or altered intracellular localization. The peptideoligonucleotide-modified nanoparticles showed a 75% decreased expression of glyceraldehyde 3-phosphate dehydrogenase, whereas oligonucleotide-modified nanoparticles showed only a 50% decrease in expression (4 ). Interestingly, the increased knockdown was not associated with greater numbers of nanoparticles per cell, but the nanoparticles were reproducibly associated more closely to the cell's nucleus.…”

Section: Peptide Antisense Nanoparticlesmentioning

confidence: 79%

“…It is against this backdrop that a series of papers out of the Mirkin group at Northwestern University is significant (1)(2)(3)(4). Few disagree with the power that oligonucleotide probes offer as detectors of nucleic acids and regulators of gene expression in living cells.…”

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confidence: 99%