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ABSTRACT: We report the development of a novel colorimetric nitrite and nitrate ion assay based upon gold nanoparticle probes functionalized with Griess reaction reagents. This assay takes advantage of the distance-dependent plasmonic properties of the gold nanoparticles and the ability of nitrite ion to facilitate the cross coupling of novel nanoparticle probes modified with aniline and naphthalene moieties. The assay works on the concept of a kinetic end point and can be triggered at the EPA limit for this ion in drinking water (highlighted in red, microM). This rapid and simple assay could be useful for on-site water quality monitoring.
Journal of the American Chemical Society 05/2009; 131(18):6362-3. · 9.91 Impact Factor
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ABSTRACT: We report the development of a highly sensitive and selective colorimetric detection method for cysteine based upon oligonucleotide-functionalized gold nanoparticle probes that contain strategically placed thymidine-thymidine (T-T) mismatches complexed with Hg2+. This assay relies upon the distance-dependent optical properties of gold nanoparticles, the sharp melting transition of oligonucleotide-linked nanoparticle aggregates, and the very selective coordination of Hg2+ with cysteine. The concentration of cysteine can be determined by monitoring with the naked eye or a UV-vis spectrometer the temperature at which the purple-to-red color change associated with aggregate dissociation takes place. This assay does not utilize organic cosolvents, enzymatic reactions, light-sensitive dye molecules, lengthy protocols, or sophisticated instrumentation thereby overcoming some of the limitations of more conventional methods.
Nano Letters 03/2008; 8(2):529-33. · 13.20 Impact Factor
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ABSTRACT: We have developed a novel competition assay that uses a gold nanoparticle (Au NP)-based, high-throughput colorimetric approach to screen the sequence selectivity of DNA-binding molecules. This assay hinges on the observation that the melting behavior of DNA-functionalized Au NP aggregates is sensitive to the concentration of the DNA-binding molecule in solution. When short, oligomeric hairpin DNA sequences were added to a reaction solution consisting of DNA-functionalized Au NP aggregates and DNA-binding molecules, these molecules may either bind to the Au NP aggregate interconnects or the hairpin stems based on their relative affinity for each. This relative affinity can be measured as a change in the melting temperature (Tm) of the DNA-modified Au NP aggregates in solution. As a proof of concept, we evaluated the selectivity of 4',6-diamidino-2-phenylindone (an AT-specific binder), ethidium bromide (a nonspecific binder), and chromomycin A (a GC-specific binder) for six sequences of hairpin DNA having different numbers of AT pairs in a five-base pair variable stem region. Our assay accurately and easily confirmed the known trends in selectivity for the DNA binders in question without the use of complicated instrumentation. This novel assay will be useful in assessing large libraries of potential drug candidates that work by binding DNA to form a drug/DNA complex.
Analytical Chemistry 10/2007; 79(18):7201-5. · 5.86 Impact Factor
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ABSTRACT: We have designed a chip-based assay, using microarray technology, for determining the relative binding affinities of duplex and triplex DNA binders. This assay combines the high discrimination capabilities afforded by DNA-modified Au nanoparticles with the high-throughput capabilities of DNA microarrays. The detection and screening of duplex DNA binders are important because these molecules, in many cases, are potential anticancer agents as well as toxins. Triplex DNA binders are also promising drug candidates. These molecules, in conjunction with triplex-forming oligonucleotides, could potentially be used to achieve control of gene expression by interfering with transcription factors that bind to DNA. Therefore, the ability to screen for these molecules in a high-throughput fashion could dramatically improve the drug screening process. The assay reported here provides excellent discrimination between strong, intermediate, and weak duplex and triplex DNA binders in a high-throughput fashion.
Analytical Chemistry 09/2007; 79(15):6037-41. · 5.86 Impact Factor
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Angewandte Chemie International Edition 02/2007; 46(22):4093-6. · 13.45 Impact Factor
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Angewandte Chemie International Edition 02/2007; 46(19):3468-70. · 13.45 Impact Factor
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ABSTRACT: We describe the use of gold nanoparticle-oligonucleotide complexes as intracellular gene regulation agents for the control of protein expression in cells. These oligonucleotide-modified nanoparticles have affinity constants for complementary nucleic acids that are higher than their unmodified oligonucleotide counterparts, are less susceptible to degradation by nuclease activity, exhibit greater than 99% cellular uptake, can introduce oligonucleotides at a higher effective concentration than conventional transfection agents, and are nontoxic to the cells under the conditions studied. By chemically tailoring the density of DNA bound to the surface of gold nanoparticles, we demonstrated a tunable gene knockdown.
Science 06/2006; 312(5776):1027-30. · 31.20 Impact Factor
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ABSTRACT: Nanoparticle assemblies interconnected with DNA triple helixes can be used to colorimetrically screen for triplex DNA binding molecules and simultaneously determine their relative binding affinities based on melting temperatures. Nanoparticles assemble only when DNA triple helixes form between DNA from two different particles and a third strand of free DNA. In addition, the triple helix structure is unstable at room temperature and only forms in the presence of triplex DNA binding molecules which stabilize the triple helix. The resulting melting transition of the nanoparticle assembly is much sharper and at a significantly higher Tm than the analogous triplex structure without nanoparticles. Upon nanoparticle assembly, a concomitant red-to-blue color change occurs. The assembly process and color change do not occur in the presence of duplex DNA binders and therefore provide a significantly better screening process for triplex DNA binding molecules compared to standard methods.
Journal of the American Chemical Society 05/2006; 128(15):4954-5. · 9.91 Impact Factor
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Angewandte Chemie International Edition 04/2006; 45(11):1807-10. · 13.45 Impact Factor
