Oligonucleotide-Modified Gold Nanoparticles for Intracellular Gene Regulation

Rosi, Nathaniel L.; Giljohann, David A.; Thaxton, C. Shad; Lytton-Jean, Abigail K. R.; Han, Min Su; Mirkin, Chad A.

doi:10.1126/science.1125559

Cited by 1,834 publications

(1,380 citation statements)

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“…6 To probe how enzymatic degradation leads to non-specific signaling, we incubated nano-flares with the endonuclease DNAse I (0.38 mg/ L, significantly greater than what would be found in a cellular environment), and measured the rate of degradation by monitoring the increase in fluorescence signal as a function of time. The results of the assay reveal that the nano-flare is degraded at a normalized rate of 0.275 nmol min −1 under these conditions.…”

Section: Introductionmentioning

confidence: 99%

“…Cell-lines transfected with nano-flares showed uniform single populations of fluorescent cells, consistent with the greater than 99% cell penetration that we observe when transfecting antisense particles. 6 Flow-cytometry revealed that SKBR3 cells treated with surviving nano-flares were highly fluorescent and 2.5 times more fluorescent than the population treated with noncomplementary controls (Figure 2). For comparison, in C166 cell models, both probes resulted in a similarly low fluorescent signal.…”

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confidence: 99%

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Nano-Flares: Probes for Transfection and mRNA Detection in Living Cells

et al. 2007

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We demonstrate that novel oligonucleotide-modified gold nanoparticle probes hybridized to fluorophore-labeled complements can be used as both transfection agents and cellular "nanoflares" for detecting mRNA in living cells. Nano-flares take advantage of the highly efficient fluorescence quenching properties of gold, cellular uptake of oligonucleotide nanoparticle conjugates without the use of transfection agents, and the enzymatic stability of such conjugates, thus overcoming many of the challenges to creating sensitive and effective intracellular probes. Nano-flares exhibit high signaling, have low background fluorescence, and are sensitive to changes in the number of RNA transcripts present in cells.Probes to visualize and detect intracellular RNA including those used for in situ staining, 1 molecular beacons, 2 and fluorescent resonance energy transfer (FRET) pairs 3 are important tools to measure and quantify activity in living systems in response to external stimuli. 4 However, these probes are often difficult to transfect, require additional agents for cellular internalization, and can be unstable in cellular environments. These factors can lead to a high background signal and the inability to detect targets. Here we show how novel oligonucleotide-modified gold nanoparticle probes hybridized to fluorophore complements can be used as both transfection agents and cellular "nano-flares" for visualizing and quantifying RNA in living cells. Nano-flares take advantage of the highly efficient fluorescence quenching properties of gold, 5 cellular uptake of oligonucleotide nanoparticle conjugates without the use of transfection agents, and the enzymatic stability of such conjugates, 6 thus overcoming many of the challenges to creating sensitive and effective intracellular probes. Specifically, nano-flares exhibit high signaling, have low background fluorescence, and are sensitive to changes in the number of RNA transcripts present in cells. The discovery and subsequent development of the oligonucleotide-nanoparticle conjugate have led to a variety of new opportunities in molecular diagnostics 11 and materials design. 12 Recently, it has been demonstrated that oligonucleotide-functionalized nanoparticles enter cells and can act as antisense agents to control gene expression. 6 These "antisense particles" are not only delivery vehicles, 13 but also single entity regulation and transfection agents that undergo facile cellular internalization, resist enzymatic degradation, and bind intracellular targets with affinity constants that are as much as two orders of magnitude greater than free oligonucleotides. 14 Moreover, they can be easily modified with potent designer materials such as locked nucleic acids 15 and are nontoxic under conditions required for gene regulation.The nano-flares described herein are oligonucleotide functionalized nanoparticle conjugates designed to provide an intracellular fluorescence signal that correlates with the relative amount of a specific intracellular RNA. By utilizing nanopart...

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Section: Introductionmentioning

confidence: 99%

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confidence: 99%

Nano-Flares: Probes for Transfection and mRNA Detection in Living Cells

et al. 2007

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“…Detailed studies have been performed for these materials and the findings might be of importance for the pristine DNA assemblies as well. By incubation of DNA-Au NPs with DNAse I it was demonstrated that ds nucleic acids are more stable when bound to a core compared to pristine ds DNA [27]. The same stability was observed in an intracellular environment (C166 cells).…”

Section: Dna-au Npsmentioning

confidence: 77%

“…DNA-Au NPs offer some extra features like magnetic properties, plasmonic effects or the ability of fluorescence quenching, which represent a significant extension to the functionality of pristine DNA nanoobjects [24]. These characteristics are important in the field of bio-imaging and biomedicine; hence DNA-Au NPs have become a very popular template for the assembly of nanoobjects and currently find use in imaging, detection and as transfection agents and gene regulation materials [25][26][27][28].…”

Section: Introductionmentioning

confidence: 99%

Drug delivery systems based on nucleic acid nanostructures

Vries

Zhang

Herrmann

2013

Journal of Controlled Release

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a b s t r a c t a r t i c l e i n f oThe field of DNA nanotechnology has progressed rapidly in recent years and hence a large variety of 1D-, 2D-and 3D DNA nanostructures with various sizes, geometries and shapes is readily accessible. DNA-based nanoobjects are fabricated by straight forward design and self-assembly processes allowing the exact positioning of functional moieties and the integration of other materials. At the same time some of these nanosystems are characterized by a low toxicity profile. As a consequence, the use of these architectures in a biomedical context has been explored. In this review the progress and possibilities of pristine nucleic acid nanostructures and DNA hybrid materials for drug delivery will be discussed. For the latter class of structures, a distinction is made between carriers with an inorganic core composed of gold or silica and amphiphilic DNA block copolymers that exhibit a soft hydrophobic interior.

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“…Dual delivery of multiple growth factors and morphogens has been achieved in multiple systems including poly (lactide-co-glycolide) (PLG) [31], oligo(poly(ethylene glycol) fumarate) [33], and alginate [34] scaffolds. Multiple factors can be delivered sequentially, simultaneously, or both, and these techniques can be combined with DNA or RNAi delivery to enable long-standing protein expression [35] or abrogation of expression [36]. The physiologic relevance of control over factors availability in time and space has been demonstrated by differentiation of stem cells [37], regulation over the extent of vascular network maturation with time [31], enhanced delivery and integration of transplanted cells [38], and the simultaneous formation of vascular networks at distinct stages of maturation in distinct spatial compartments [28].…”

Section: Spatial and Temporal Control Of Morphogen And Growth Factormentioning

confidence: 99%

Polymers to direct cell fate by controlling the microenvironment

Sands

Mooney

2007

Current Opinion in Biotechnology

130

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Enhanced understanding of the signals within the microenvironment that regulate cell fate has led to the development of increasingly sophisticated polymeric biomaterials for tissue engineering and regenerative medicine applications. This advancement is exemplified by biomaterials with precisely controlled scaffold architecture, that regulate the spatio-temporal release of growth factors and morphogens, and respond dynamically to microenvironmental cues. Further understanding of the biology, qualitatively and quantitatively, of cells within their microenvironments and at the tissuematerial interface will expand the design space of future biomaterials.

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Oligonucleotide-Modified Gold Nanoparticles for Intracellular Gene Regulation

Cited by 1,834 publications

References 30 publications

Nano-Flares: Probes for Transfection and mRNA Detection in Living Cells

Nano-Flares: Probes for Transfection and mRNA Detection in Living Cells

Drug delivery systems based on nucleic acid nanostructures

Polymers to direct cell fate by controlling the microenvironment

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