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WIREs Nanomed Nanobiotechnol
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Noble metal nanoparticles in DNA detection and delivery

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DNA‐conjugated metal nanoparticles have attracted enormous attention for biological and medical applications, owing to their unusual DNA melting characteristics as well as unique optical and catalytic properties. The combination of these unique properties has not only led to the development of DNA‐detection technologies with remarkably high selectivity and sensitivity, but also to the development of gene therapeutic agents with high efficacy and efficiency. In this review, we present a comprehensive coverage on their applications in detecting, manipulating, and delivering genes. WIREs Nanomed Nanobiotechnol 2012, 4:273–290. doi: 10.1002/wnan.1159

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Figure 1.

(a) A transmission electron microscopy (TEM) image, an extinction spectrum, and a picture of 13‐nm gold nanosphere solution. (b) TEM images of silica particles coated with different amounts of gold for the formation of a gold nanoshell. Extinction spectra of nanoshells with different core to shell ratios show that the surface plasmon resonance (SPR) band of nanoshells red‐shifts with decreasing shell thicknesses. (c) A TEM image and extinction spectra of gold nanorods. Extinction spectra of nanorods with different aspect ratios show that the longitudinal SPR band of nanorods red‐shifts with increasing aspect ratios. (d) A TEM image and an extinction spectrum of gold nanostars. (e) TEM images of spiky gold nanoshells with spherical and rod cores and an extinction spectrum of spiky gold nanoshells made with spherical cores. (f) A TEM image of gold nanoprisms and extinction spectra of nanoprisms with different edge lengths. (g) TEM images and extinction spectra of silver nanocubes and gold nanocages. (h) Scanning electron microscope (left) and optical microscope (right) images of a multicomponent nanowire. (i) Three different sized small gold clusters with UV (Au5), blue (Au8), and green (Au13) emission under UV irradiation and their corresponding excitation (dotted) and emission (solid) spectra (b: Reprinted with permission from Ref 22. Copyright 2006 Springer Science and Business Media; c: Reprinted with permission from Ref 23. Copyright 2009 Wiley‐VCH Verlag GmbH & Co. KGaA; d: Reprinted with permission from Ref 24. Copyright 2006 American Chemical Society; f: Reprinted with permission from Ref 25. Copyright 2006 Wiley‐VCH Verlag GmbH & Co. KGaA.; g: Reprinted with permission from Ref 26. Copyright 2007 Wiley‐VCH Verlag GmbH & Co. KGaA; h: Reprinted with permission from Ref 27. Copyright 2004 American Physical Society; i: Reprinted with permission from Ref 28. Copyright 2004 AAAS.)

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Figure 2.

Preparation of nanoparticles densely functionalized with DNA. (a) A widely used conjugation method using citrate‐stabilized gold nanoparticles and thiol‐modified DNA. (b) DNA dissociation curves of DNA‐modified nanoparticles and unlabeled DNA, showing that the nanoparticle probes exhibit unusually sharp melting transitions. (c) A DNA‐functionalization method based on the self‐assembly of DNA block‐copolymers and nanoparticles. This method can be used to coat different types of nanoparticles with a dense layer of DNA.

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Figure 3.

Colorimetric DNA‐detection methods using DNA‐modified gold nanoparticles (a) and unmodified gold nanoparticles (b, c). These methods depend on the change in color [e.g., shift in the surface plasmon resonance (SPR) band] upon nanoparticle aggregation or de‐aggregation. (a) The presence of target DNA strands that are complementary to the DNA strands immobilized on gold nanoparticles results in the formation of nanoparticle aggregates and red to purple color change. (b) A label‐free colorimetric detection method using citrate‐stabilized gold nanoparticles. In the absence of target DNA, the binding of ssDNA on citrate‐stabilized gold nanoparticles stabilizes the particles in salt solutions. When dsDNA containing target DNA was added, on the other hand, nanoparticles aggregate in salt solutions, causing red to purple color change. (c) A label‐free colorimetric detection using peptide nucleic acid (PNA). Citrate‐stabilized gold nanoparticles aggregate upon the addition of PNA. Subsequent addition of complementary DNA targets redisperses nanoparticles, causing a purple to red color change. (b: Pictures of the nanoparticle solutions with ssDNA and dsDNA after the addition of salt; Reprinted with permission from Ref 72. Copyright 2004 National Academy of Sciences, USA.)

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Figure 4.

Schematic representations for the detection of DNA using the scanometric (a) and electrical (b) methods. The two methods are similar in design with the major difference in the readout step, where the results of the scanometric method can be detected by a scanner or the naked eye and the electrical method provides an electrical signal.

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Figure 5.

(a) Schematic description of chip‐based DNA detection using surface enhanced Raman spectroscopy (SERS). The SERS signal from reporter dyes is increased by the catalytic silver deposition on nanoparticles. Different reporter dye molecules show different SERS spectra, providing this method the capability for multiplexing. (b) A homogeneous DNA‐detection method based on SERS. SERS signal is increased in the presence of target DNA because Raman enhancement factor is higher in nanoparticle aggregates than in isolated nanoparticles. (c) Scheme and transmission electron microscopy (TEM) image of an Au–Ag core‐shell nanoparticle dimer prepared from DNA‐functionalized gold nanoparticles. (d) SERS spectra of Au–Ag core‐shell dimers with 3 nm (top) and 10 nm (bottom) thick Ag shell, showing that the dimer with a narrower gap yields higher SERS signal. (e) A diagram for a DNA‐detection scheme based on surface plasmon resonance spectroscopy (SPRS). (f) SPRS spectra showing that the binding of gold nanoparticles through DNA hybridization results in a large shift in SPR positions. (c and d: Reprinted with permission from Ref 87. Copyright 2010 AAAS; f: Reprinted with permission from Ref 88. Copyright 2000 American Chemical Society.)

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Figure 6.

(a–c) Gene delivery and regulation using DNA‐functionalized gold nanospheres. In this method, antisense DNA strands that inhibit the expression of enhanced green fluorescent protein (EGFP) were first immobilized on gold probes. Cy5‐modified reporter DNA strands, which were used to monitor the cellular uptake, were hybridized to the antisense DNA and then the nanoparticle probes were incubated with C166‐EGFP cells. Confocal fluorescence microscopy images of C166‐EGFP cells before and after the treatment with the nanoparticle probes are shown in (b) and (c), respectively. (upper left: Cy5 emission; upper right: EGFP emission; lower left: transmission image; lower right: overlay of the three channels). Results indicate that the reporter–antisense probes were effectively taken up by cells (appearance of red fluorescence from Cy5 in (c)) and reduced the expression of EGFP in the cells (reduced green fluorescence from EGFP in (c)). (d–e) Intracellular RNA detection method based on DNA‐modified nanoparticles called ‘nano‐flares’. In this work, Cy5‐modified reporter DNA strands were hybridized to the Survivin antisense DNA on nanoparticles in a way that Cy5 dyes were close to the gold surface. In that geometry, fluorescence of Cy5 was completely quenched by gold. In the presence of mRNA, the short Cy5 labeled reporter strands are dehybridized from the nano‐flares and therefore become fluorescent (left image in (e)). Shown in (e) are the cells treated with nano‐flares (left: cells containing survivin; right: cells without survivin). Shown in (f) is another control where cells are treated with non‐survivin antisense nano‐flares (left: cells containing survivin; right: cells without survivin); neither show any fluorescence from the reporter. (b and c: Reprinted with permission from Ref 103. Copyright 2006 AAAS; (e) and (f): Reprinted with permission from Ref 105. Copyright 2007 American Chemical Society.)

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Figure 7.

(a) Release of DNA strands hybridized to complementary DNA immobilized on gold nanoshells by near infrared (NIR) irradiation. (b) Release of green fluorescent protein (GFP) coding plasmid from Au nanorod through the shape transformation by pulsed laser irradiation. (c) Experimental setup and schematic description of radio‐frequency (RF)‐field‐induced DNA dehybridization.

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In the Spotlight

James F. Leary

James F. Leary
has been contributing to nanomedical research and technologies throughout his career. Such contributions include the invention of high-speed flow cytometry, cell sorting techniques, and rare-event methods. Dr. Leary’s current research spans across three general areas in nanomedicine. The first is the development of high-throughput single-cell flow cytometry and cell sorting technologies. The second explores BioMEMS technologies. These include miniaturized cell sorters, portable devices for detection of microbial pathogens in food and water, and artificial human “organ-on-a-chip” technologies which consists of developing cell culture chips capable of simulating the activities and mechanics of entire organs and organ systems. His third area of research aims at developing smart nano-engineered systems for single-cell drug or gene delivery for nanomedicine. Dr. Leary currently holds nine issued U.S. Patents with four currently pending, and he has received NIH funding for over 25 years.

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