CiteULike: Tag origami

Origami constructions

Antonio Oller — 2007-09-21T08:10:42-00:00

(20 Sep 2007)

characterization of real numbers constructible by paper folding.

Folding DNA to create nanoscale shapes and patterns

Paul Rothemund — 2006-03-15T23:36:35-00:00

Nature, Vol. 440, No. 7082., pp. 297-302.

Rapid synthesis of DNA-cysteine conjugates for expressed protein ligation

Marina Lovrinovic — 2008-02-20T06:52:24-00:00

Biochemical and Biophysical Research Communications, Vol. 335, No. 3. (30 September 2005), pp. 943-948.

We report a rapid method for the covalent modification of commercially available amino-modified DNA oligonucleotides with a cysteine moiety. The resulting DNA-cysteine conjugates are versatile reagents for the efficient preparation of covalent DNA-protein conjugates by means of expressed protein ligation (EPL). The EPL method allows for the site-specific coupling of cysteine-modified DNA oligomers with recombinant intein-fusion proteins, the latter of which contain a C-terminal thioester enabling the mild and highly specific reaction with N-terminal cysteine compounds. We prepared a cysteine-modifier reagent in a single-step reaction which allows for the rapid and near quantitative synthesis of cysteine-DNA conjugates. The latter were ligated with the green fluorescent protein mutant EYFP, recombinantly expressed as an intein-fusion protein, allowing for the mild and selective formation of EYFP-DNA conjugates in high yields of about 60%. We anticipate many applications of our approach, ranging from protein microarrays to the arising field of nanobiotechnology.

DNA-programmable nanoparticle crystallization

Sung Park — 2008-01-31T08:37:22-00:00

Nature, Vol. 451, No. 7178. (31 January 2008), pp. 553-556.

DNA-guided crystallization of colloidal nanoparticles

Dmytro Nykypanchuk — 2008-01-31T08:32:06-00:00

Nature, Vol. 451, No. 7178. (31 January 2008), pp. 549-552.

From DNA to transistors

E Braun — 2008-05-06T11:33:46-00:00

Advances in Physics, Vol. 53 (June 2004), pp. 441-496.

The rapid advance in molecular biology and nanotechnology opens up the possibility to explore the interface between biology and electronics at the single-molecule level. We focus on the organization of molecular electronic circuits. Interconnecting an immense number of molecular devices into a functional circuit and constructing a framework for integrated molecular electronics requires new concepts. A promising avenue relies on bottom-up assembly where the information for the circuit connectivity and functionality is embedded in the molecular building blocks. Biology can provide concepts and mechanisms for advancing this approach, but there is no straightforward way to apply them to electronics since biological molecules are essentially electrically insulating. Bridging the chasm between biology and electronics therefore presents great challenges. Circuit organization on the molecular scale is considered and contrasted with the levels of organization presented by the living world. The discussion then focuses on our proposal to harness DNA and molecular biology to construct the scaffold for integrated molecular electronics. DNA metallization is used to convert the DNA scaffold into a conductive one. We present the framework of sequence-specific molecular lithography based on the biological mechanism of homologous genetic recombination and carried out by the bacterial protein RecA. Molecular lithography enables us to use the information encoded in the scaffold DNA molecules for directing the construction of an electronic circuit. We show that it can lead all the way from DNA molecules to working transistors in a test-tube. Carbon nanotubes are incorporated as the active electronic components in the DNA-templated transistors. Our approach can, in principle, be applied to the fabrication of larger-scale electronic circuits. The realization of complex DNA-based circuits will, however, require new concepts and additional biological machinery allowing, for example, feedback from the electronic functionality to direct the assembly process and adaptation mechanisms.

DNA-Directed Immobilization: Efficient, Reversible, and Site-Selective Surface Binding of Proteins by Means of Covalent DNA-Streptavidin Conjugates

Christof Niemeyer — 2008-02-20T06:49:43-00:00

Analytical Biochemistry, Vol. 268, No. 1. (1 March 1999), pp. 54-63.

Covalent DNA-streptavidin conjugates have been utilized for the reversible and site-selective immobilization of various biotinylated enzymes and antibodies by DNA-directed immobilization (DDI). Biotinylated alkaline phosphatase, [beta]-galactosidase, and horseradish peroxidase as well as biotinylated anti-mouse and anti-rabbit immunoglobulins have been coupled to the DNA-streptavidin adapters by simple, two-component incubation and the resulting preconjugates were allowed to hybridize to complementary, surface-bound capture oligonucleotides. Quantitative measurements on microplates indicate that DDI proceeds with a higher immobilization efficiency than conventional immobilization techniques, such as the binding of the biotinylated proteins to streptavidin-coated surfaces or direct physisorption. These findings can be attributed to the reversible formation of the rigid, double-stranded DNA spacer between the surface and the proteins. Moreover, BIAcore measurements demonstrate that DDI allows a reversible functionalization of sensor surfaces with reproducible amounts of proteins. Ultimately, the simultaneous immobilization of different compounds using microstructured oligonucleotide arrays as immobilization matrices demonstrate that DDI proceeds with site selectivity due to the unique specificity of Watson-Crick base pairing.

Design and characterization of two-dye and three-dye binary fluorescent probes for mRNA detection

Angel Marti — 2008-03-13T09:25:39-00:00

Tetrahedron, Vol. 63, No. 17. (23 April 2007), pp. 3591-3600.

We report the design, synthesis, and characterization of binary oligonucleotide probes for mRNA detection. The probes were designed to avoid common problems found in standard binary probes such as direct excitation of the acceptor fluorophore and overlap between the donor and acceptor emission spectra. Two different probes were constructed that contained an array of either two or three dyes and were characterized using steady-state fluorescence spectroscopy, time-resolved fluorescence spectroscopy, and fluorescence depolarization measurements. The three-dye binary probe (BP-3d) consists of a Fam fluorophore which acts as a donor, collecting light and transferring it as energy to Tamra, which subsequently transfers energy to Cy5 when the two probes are hybridized to mRNA. This design allows the use of 488 nm excitation, which avoids the direct excitation of Cy5 and at the same time provides a good fluorescence resonance energy transfer (FRET) efficiency. The two-dye binary probe system (BP-2d) was constructed with Alexa488 and Cy5 fluorophores. Although the overlap between the fluorescence of Alexa488 and the absorption of Cy5 is relatively low, FRET still occurs due to their close physical proximity when the probes are hybridized to mRNA. This framework also decreases the direct excitation of Cy5 and reduces the fluorescence overlap between the donor and the acceptor. Picosecond time-resolved spectroscopy showed a reduction in the fluorescence lifetime of donor fluorophores after the formation of the hybrid between the probes and target mRNA. Interestingly, BP-2d in the presence of mRNA shows a slow rise in the fluorescence decay of Cy5 due to a relatively slow FRET rate, which together with the reduction in the Alexa488 lifetime provides a way to improve the signal to background ratio using time-resolved fluorescence spectra (TRES). In addition, fluorescence depolarization measurements showed complete depolarization of the acceptor dyes (Cy5) for both BP-3d (due to sequential FRET steps) and BP-2d (due to the relatively low FRET rate) in the presence of the mRNA target.

Simple model for DNA adsorption onto a mica surface in 1:1 and 2:1 electrolyte solutions.

ML Sushko — 2007-12-12T15:19:43-00:00

Langmuir, Vol. 22, No. 18. (29 August 2006), pp. 7678-7688.

We propose a simple theory of interactions between like-charged polyelectrolyte and a surface based on a mean-field Derjaguin-Landau-Verwey-Overbeek approach. It predicts that the van der Waals attractive interactions are responsible for irreversible physisorption of polyelectrolytes onto charged surfaces. We show that monovalent salts contribute significantly to repulsive interactions, while enhancing the attraction very slightly. The effect of the divalent counterions is reverse. Therefore, to achieve the adsorption, the overall repulsion due to 1:1 electrolyte should be counterbalanced by the stronger van der Waals attraction due to the presence of doubly charged counterions in solution. The theory has been validated experimentally against its ability to predict the minimum polymer/surface interaction energy required for the adsorption using DNA/mica in NaCl, MgCl2, and NiCl2 solutions as a test system. The theory explains the mechanism of linear DNA adsorption to a mica surface for different solvent compositions and can be used as a tool for predicting the optimum conditions for AFM experiments on linear polymer systems. The model can also be used to make general conclusions on the conformation of polymer molecules on a surface. We have shown for the DNA/mica surface system that when the adsorption of DNA is mostly governed by long-range van der Waals forces the molecule adopts an ideal 2D conformation. When the adsorption is mostly due to short-range ion-correlation forces, DNA will appear 3D --> 2D projected in agreement with experimental data.

Sequence-Specific Molecular Lithography on Single DNA Molecules

Kinneret Keren — 2008-02-20T09:28:53-00:00

Science, Vol. 297, No. 5578. (5 July 2002), pp. 72-75.

10.1126/science.1071247

Nanoparticles, Proteins, and Nucleic Acids: Biotechnology Meets Materials Science

Christof Niemeyer — 2007-05-09T19:34:06-00:00

Angewandte Chemie International Edition, Vol. 40, No. 22. (2001), pp. 4128-4158.

Based on fundamental chemistry, biotechnology and materials science have developed over the past three decades into today's powerful disciplines which allow the engineering of advanced technical devices and the industrial production of active substances for pharmaceutical and biomedical applications. This review is focused on current approaches emerging at the intersection of materials research, nanosciences, and molecular biotechnology. This novel and highly interdisciplinary field of chemistry is closely associated with both the physical and chemical properties of organic and inorganic nanoparticles, as well as to the various aspects of molecular cloning, recombinant DNA and protein technology, and immunology. Evolutionary optimized biomolecules such as nucleic acids, proteins, and supramolecular complexes of these components, are utilized in the production of nanostructured and mesoscopic architectures from organic and inorganic materials. The highly developed instruments and techniques of today's materials research are used for basic and applied studies of fundamental biological processes.

Covalent cross-linking of vasoactive intestinal polypeptide to its receptors on intact human lymphoblasts

CL Wood — 2008-01-22T16:26:35-00:00

J. Biol. Chem., Vol. 260, No. 2. (25 January 1985), pp. 1243-1247.

Nanoscale structure and dynamics of DNA.

MA Berg — 2008-05-06T17:47:19-00:00

Physical chemistry chemical physics : PCCP, Vol. 10, No. 9. (7 March 2008), pp. 1229-1242.

DNA is depicted in elementary chemistry and biology texts as a perfect double helix; but local structural variations and nanoscale motions within the double helix are critical for its ability to be packaged, recognized, and transcribed. DNA is becoming a favored nanoscale assembly tool due to the precise pairing of complementary strands that in principle can bring nanoscale objects within a well-defined distance of each other. However, future nanotechnology applications of DNA need to take into account its variable nanoscale structural and dynamic properties, especially in terms of its solvent shell and counterions. This article highlights efforts of the authors to (1) interrogate nanoscale structures of DNA using nanoparticles and (2) measure the dynamic nature of DNA over six orders of magnitude in time, using a fluorescent reporter in the base stack.

Adsorption of DNA to mica mediated by divalent counterions: a theoretical and experimental study.

D Pastré — 2007-12-12T15:17:15-00:00

Biophys J, Vol. 85, No. 4. (October 2003), pp. 2507-2518.

The adsorption of DNA molecules onto a flat mica surface is a necessary step to perform atomic force microscopy studies of DNA conformation and observe DNA-protein interactions in physiological environment. However, the phenomenon that pulls DNA molecules onto the surface is still not understood. This is a crucial issue because the DNA/surface interactions could affect the DNA biological functions. In this paper we develop a model that can explain the mechanism of the DNA adsorption onto mica. This model suggests that DNA attraction is due to the sharing of the DNA and mica counterions. The correlations between divalent counterions on both the negatively charged DNA and the mica surface can generate a net attraction force whereas the correlations between monovalent counterions are ineffective in the DNA attraction. DNA binding is then dependent on the fractional surface densities of the divalent and monovalent cations, which can compete for the mica surface and DNA neutralizations. In addition, the attraction can be enhanced when the mica has been pretreated by transition metal cations (Ni(2+), Zn(2+)). Mica pretreatment simultaneously enhances the DNA attraction and reduces the repulsive contribution due to the electrical double-layer force. We also perform end-to-end distance measurement of DNA chains to study the binding strength. The DNA binding strength appears to be constant for a fixed fractional surface density of the divalent cations at low ionic strength (I < 0.1 M) as predicted by the model. However, at higher ionic strength, the binding is weakened by the screening effect of the ions. Then, some equations were derived to describe the binding of a polyelectrolyte onto a charged surface. The electrostatic attraction due to the sharing of counterions is particularly effective if the polyelectrolyte and the surface have nearly the same surface charge density. This characteristic of the attraction force can explain the success of mica for performing single DNA molecule observation by AFM. In addition, we explain how a reversible binding of the DNA molecules can be obtained with a pretreated mica surface.

Double-labeled donor probe can enhance the signal of fluorescence resonance energy transfer (FRET) in detection of nucleic acid hybridization

Yukio Okamura — 2008-03-13T09:22:41-00:00

Nucl. Acids Res., Vol. 28, No. 24. (15 December 2000), e107.

A set of fluorescently-labeled DNA probes that hybridize with the target RNA and produce fluorescence resonance energy transfer (FRET) signals can be utilized for the detection of specific RNA. We have developed probe sets to detect and discriminate single-strand RNA molecules of plant viral genome, and sought a method to improve the FRET signals to handle in vivo applications. Consequently, we found that a double-labeled donor probe labeled with Bodipy dye yielded a remarkable increase in fluorescence intensity compared to a single-labeled donor probe used in an ordinary FRET. This double-labeled donor system can be easily applied to improve various FRET probes since the dependence upon sequence and label position in enhancement is not as strict. Furthermore this method could be applied to other nucleic acid substances, such as oligo RNA and phosphorothioate oligonucleotides (S-oligos) to enhance FRET signal. Although the double-labeled donor probes labeled with a variety of fluorophores had unexpected properties (strange UV-visible absorption spectra, decrease of intensity and decay of donor fluorescence) compared with single-labeled ones, they had no relation to FRET enhancement. This signal amplification mechanism cannot be explained simply based on our current results and knowledge of FRET. Yet it is possible to utilize this double-labeled donor system in various applications of FRET as a simple signal-enhancement method. 10.1093/nar/28.24.e107

Organization of 'nanocrystal molecules' using DNA

Paul Alivisatos — 2007-09-27T18:37:01-00:00

Nature, Vol. 382, No. 6592. (1996), pp. 609-611.

PATTERNING matter on the nanometre scale is an important objective of current materials chemistry and physics. It is driven by both the need to further miniaturize electronic components and the fact that at the nanometre scale, materials properties are strongly size-dependent and thus can be tuned sensitively1. In nanoscale crystals, quantum size effects and the large number of surface atoms influence the, chemical, electronic, magnetic and optical behaviour2—4. 'Top-down' (for example, lithographic) methods for nanoscale manipulation reach only to the upper end of the nanometre regime5; but whereas 'bottom-up' wet chemical techniques allow for the preparation of mono-disperse, defect-free crystallites just 1–10 nm in size6–10, ways to control the structure of nanocrystal assemblies are scarce. Here we describe a strategy for the synthesis of'nanocrystal molecules', in which discrete numbers of gold nanocrystals are organized into spatially defined structures based on Watson-Crick base-pairing interactions. We attach single-stranded DNA oligonucleotides of defined length and sequence to individual nanocrystals, and these assemble into dimers and trimers on addition of a complementary single-stranded DNA template. We anticipate that this approach should allow the construction of more complex two-and three-dimensional assemblies.

High-Quality Mapping of DNA-Protein Complexes by Dynamic Scanning Force Microscopy

Song Gao — 2008-07-09T13:44:23-00:00

ChemPhysChem, Vol. 2, No. 6. (2001), pp. 384-388.

No Abstract

Gold nanoparticle aggregation-based highly sensitive DNA detection using atomic force microscopy.

MP Bui — 2008-02-20T10:04:38-00:00

Anal Bioanal Chem, Vol. 388, No. 5-6. (July 2007), pp. 1185-1190.

The potential ability of atomic force microscopy (AFM) as a quantitative bioanalysis tool is demonstrated by using gold nanoparticles as a size enhancer in a DNA hybridization reaction. Two sets of probe DNA were functionalized on gold nanoparticles and sandwich hybridization occurred between two probe DNAs and target DNA, resulting in aggregation of the nanoparticles. At high concentrations of target DNA in the range from 100 nM to 10 microM, the aggregation of gold nanoparticles was determined by monitoring the color change with UV-vis spectroscopy. The absorption spectra broadened after the exposure of DNA-gold nanoparticles to target DNA and a new absorption band at wavelengths >600 nm was observed. However, no differences were observed in the absorption spectra of the gold nanoparticles at low concentrations of target DNA (10 pM to 10 nM) due to insufficient aggregation. AFM was used as a biosensing tool over this range of target DNA concentrations in order to monitor the aggregation of gold nanoparticles and to quantify the concentration of target DNA. Based on the AFM images, we successfully evaluated particle number and size at low concentrations of target DNA. The calibration curve obtained when mean particle aggregate diameter was plotted against concentration of target DNA showed good linearity over the range 10 pM to 10 nM, the working range for quantitative target DNA analysis. This AFM-based DNA detection technique was three orders of magnitude more sensitive than a DNA detection method based on UV-vis spectroscopy.

Hierarchical self-assembly of DNA into symmetric supramolecular polyhedra

Yu He — 2008-03-13T00:31:06-00:00

Nature, Vol. 452, No. 7184. (13 March 2008), pp. 198-201.

Nanostructures in biodiagnostics.

NL Rosi — 2005-09-21T14:12:47-00:00

Chem Rev, Vol. 105, No. 4. (April 2005), pp. 1547-1562.

3DNA: a software package for the analysis, rebuilding and visualization of three-dimensional nucleic acid structures.

XJ Lu — 2006-05-09T20:53:54-00:00

Nucleic Acids Res, Vol. 31, No. 17. (1 September 2003), pp. 5108-5121.

We present a comprehensive software package, 3DNA, for the analysis, reconstruction and visualization of three-dimensional nucleic acid structures. Starting from a coordinate file in Protein Data Bank (PDB) format, 3DNA can handle antiparallel and parallel double helices, single-stranded structures, triplexes, quadruplexes and other complex tertiary folding motifs found in both DNA and RNA structures. The analysis routines identify and categorize all base interactions and classify the double helical character of appropriate base pair steps. The program makes use of a recently recommended reference frame for the description of nucleic acid base pair geometry and a rigorous matrix-based scheme to calculate local conformational parameters and rebuild the structure from these parameters. The rebuilding routines produce rectangular block representations of nucleic acids as well as full atomic models with the sugar-phosphate backbone and publication quality 'standardized' base stacking diagrams. Utilities are provided to locate the base pairs and helical regions in a structure and to reorient structures for effective visualization. Regular helical models based on X-ray diffraction measurements of various repeating sequences can also be generated within the program.

A DNA-based method for rationally assembling nanoparticles into macroscopic materials

Chad Mirkin — 2007-03-14T19:31:29-00:00

Nature, Vol. 382, No. 6592. (1996), pp. 607-609.

COLLOIDAL particles of metals and semiconductors have potentially useful optical, optoelectronic and material properties1–4 that derive from their small (nanoscopic) size. These properties might lead to applications including chemical sensors, spectro-scopic enhancers, quantum dot and nanostructure fabrication, and microimaging methods2–4. A great deal of control can now be exercised over the chemical composition, size and polydis-persity1,2 of colloidal particles, and many methods have been developed for assembling them into useful aggregates and materials. Here we describe a method for assembling colloidal gold nanoparticles rationally and reversibly into macroscopic aggregates. The method involves attaching to the surfaces of two batches of 13-nm gold particles non-complementary DNA oligo-nucleotides capped with thiol groups, which bind to gold. When we add to the solution an oligonucleotide duplex with 'sticky ends' that are complementary to the two grafted sequences, the nanoparticles self-assemble into aggregates. This assembly process can be reversed by thermal denaturation. This strategy should now make it possible to tailor the optical, electronic and structural properties of the colloidal aggregates by using the specificity of DNA interactions to direct the interactions between particles of different size and composition.

Fabricating and aligning pi-conjugated polymer-functionalized DNA nanowires: atomic force microscopic and scanning near-field optical microscopic studies.

H Nakao — 2008-02-20T09:59:48-00:00

Langmuir, Vol. 21, No. 17. (16 August 2005), pp. 7945-7950.

We report a simple method to functionalize DNA with pi-conjugated polymer, forming highly aligned and integrated arrays of pi-conjugated polymer nanowires of a few nanometers diameter. pi-conjugated polymer, polyphenazasiline, having alkylammonium salts on the N atom (PPhenaz-TMA), synthesized in this study can be directly attached to DNA, which can be organized along stretched and aligned DNA molecules on surfaces as a template. Furthermore, PPhenaz-TMA/DNA nanowires were stretched and aligned on surfaces, even when PPhenaz-TMA/DNA complexes formed in solutions. The resulting PPhenaz-TMA/DNA nanowires could be easily converted to oxidized states or metallic nanowires by using adequate oxidant or metal salts. The direct visualization of PPhenaz-TMA/DNA nanowires and its structural changes have been studied by atomic force microscopy and scanning near-field optical microscopy.

Dynamic interactions of p53 with DNA in solution by time-lapse atomic force microscopy

Yuekan Jiao — 2008-03-18T09:12:59-00:00

Journal of Molecular Biology, Vol. 314, No. 2. (23 November 2001), pp. 233-243.

Dynamic interactions of the tumor suppressor protein p53 with a DNA fragment containing a p53-specific recognition sequence were directly observed by time-lapse tapping mode atomic force microscopy (AFM) in liquid. The divalent cation Mg2+ was used to loosely attach both DNA and p53 to a mica surface so they could be imaged by the AFM while interacting with each other. Various interactions of p53 with DNA were observed, including dissociation/re-association, sliding and possibly direct binding to the specific sequence. Two modes of target recognition of p53 were detected: (a) direct binding, and (b) initial non-specific binding with subsequent translocation by one-dimensional diffusion of the protein along the DNA to the specific site.

Finite-Size, Fully Addressable DNA Tile Lattices Formed by Hierarchical Assembly Procedures

Sung Park — 2008-02-21T17:18:23-00:00

Angewandte Chemie, Vol. 118, No. 5. (2006), pp. 749-753.

No Abstract

AFM studies of DNA structures on mica in the presence of alkaline earth metal ions.

J Zheng — 2007-12-12T15:04:27-00:00

Biophys Chem, Vol. 104, No. 1. (1 May 2003), pp. 37-43.

As counterions of DNA on mica, Mg(2+), Ca(2+), Sr(2+) and Ba(2+) were used for clarifying whether DNA molecules equilibrate or are trapped on mica surface. End to end distance and contour lengths were determined from statistical analysis of AFM data. It was revealed that DNA molecules can equilibrate on mica when Mg(2+), Ca(2+) and Sr(2+) are counterions. When Ba(2+) is present, significantly crossovered DNA molecules indicate that it is most difficult for DNA to equilibrate on mica and the trapping degree is different under different preparation conditions. In the presence of ethanol, using AFM we have also observed the dependence of B-A conformational transition on counterion identities. The four alkaline earth metal ions cause the B-A transition in different degrees, in which Sr(2+) induces the greatest structural transition.

Nucleic acid nanostructures: bottom-up control of geometry on the nanoscale

Nadrian Seeman — 2004-12-28T15:58:29-00:00

Reports on Progress in Physics, Vol. 68, No. 1. (2005), 237.

DNA may seem an unlikely molecule from which to build nanostructures, but this is not correct. The specificity of interaction that enables DNA to function so successfully as genetic material also enables its use as a smart molecule for construction on the nanoscale. The key to using DNA for this purpose is the design of stable branched molecules, which expand its ability to interact specifically with other nucleic acid molecules. The same interactions used by genetic engineers can be used to make cohesive interactions with other DNA molecules that lead to a variety of new species. Branched DNA molecules are easy to design, and they can assume a variety of structural motifs. These can be used for purposes both of specific construction, such as polyhedra, and for the assembly of topological targets. A variety of two-dimensional periodic arrays with specific patterns have been made. DNA nanomechanical devices have been built with a series of different triggers, small molecules, nucleic acid molecules and proteins. Recently, progress has been made in self-replication of DNA nanoconstructs, and in the scaffolding of other species into DNA arrangements.

Metallic Conduction through Engineered DNA: DNA Nanoelectronic Building Blocks

A Rakitin — 2007-09-07T10:41:29-00:00

Physical Review Letters, Vol. 86, No. 16. (16 April 2001), 3670.

A novel way of engineering DNA molecules involves substituting the imino proton of each base pair with a metal ion to obtain M -DNA with altered electronic properties. We report the first direct evidence of metalliclike conduction through 15 μm long M -DNA. In contrast; measurements on B -DNA give evidence of semiconducting behavior with a few hundred meV band gap at room temperature. The drastic change of M -DNA conductivity points to a new degree of freedom in the development of future molecular electronics utilizing DNA; such as creating all-DNA junction devices for use as nanoelectronic building blocks.

Multiparameter single-molecule fluorescence spectroscopy reveals heterogeneity of HIV-1 reverse transcriptase:primer/template complexes.

PJ Rothwell — 2008-04-05T14:40:04-00:00

Proceedings of the National Academy of Sciences of the United States of America, Vol. 100, No. 4. (18 February 2003), pp. 1655-1660.

By using single-molecule multiparameter fluorescence detection, fluorescence resonance energy transfer experiments, and newly developed data analysis methods, this study demonstrates directly the existence of three structurally distinct forms of reverse transcriptase (RT):nucleic acid complexes in solution. Single-molecule multiparameter fluorescence detection also provides first information on the structure of a complex not observed by x-ray crystallography. This species did not incorporate nucleotides and is structurally distinct from the other two observed species. We determined that the nucleic acid substrate is bound at a site far removed from the nucleic acid-binding tract observed by crystallography. In contrast, the other two states are identified as being similar to the x-ray crystal structure and represent distinct enzymatically productive stages in DNA polymerization. These species differ by only a 5-A shift in the position of the nucleic acid. Addition of nucleoside triphosphate or of inorganic pyrophosphate allowed us to assign them as the educt and product state in the polymerization reaction cycle; i.e., the educt state is a complex in which the nucleic acid is positioned to allow nucleotide incorporation. The second RT:nucleic acid complex is the product state, which is formed immediately after nucleotide incorporation, but before RT translates to the next nucleotide.

A 1.4-nm gold cluster covalently attached to antibodies improves immunolabeling

JF Hainfeld — 2007-10-09T09:12:57-00:00

J. Histochem. Cytochem., Vol. 40, No. 2. (1 February 1992), pp. 177-184.

A large gold cluster (Au1.4nm) was covalently coupled to IgG and Fab' fragments. Its gold core is 1.4 nm in diameter and the Fab'-Au1.4nm immunoconjugate is the smallest gold immunoprobe that can be seen directly in the conventional electron microscope. It is useful in high- resolution immunolabeling, providing a resolution of 7.0 nm. The cluster's visibility can be enhanced with silver development for use in EM or light microscopy for histological purposes, or to detect less than or equal to 0.2 pg of antigen in immunoblots. By using a gold compound with covalent attachment, a number of advantages over colloidal gold probes are realized, including better resolution, stability, uniformity, sensitivity, and complete absence of aggregation; its small size should also improve penetration and more quantitative labeling of antigenic sites.

Method for orienting DNA molecules on mica surfaces in one direction for atomic force microscopy imaging.

M Gad — 2007-12-12T15:50:13-00:00

J Biomol Struct Dyn, Vol. 19, No. 3. (December 2001), pp. 471-477.

An efficient method was developed to stretch DNA molecules on an atomically flat surface for AFM imaging. This method involves anchoring DNA molecules from their 5' ends to amino silanized mica surfaces. N-Succinimidyl6-[3'-(2-pyridyldithio) propionamido]hexanoate (LC-SPDP), a heterobifunctional cross-linker with a flexible spacer arm was used for this purpose. Immobilization was carried out by introducing a thiol group to the 5' end of DNA by PCR. Thiolated molecules were then reacted with the cross linker to conjugate with its 2-pyridyl disulphide group via sulfhydryl exchange. The resulting complex was deposited on amino silanized mica where NHS-ester moiety of the cross linker reacted with the primary amino group on the surface. Samples were washed by a current of water and dried by an air jet in one direction parallel to the surface. DNA molecules were fully stretched in one direction on imaging them by AFM.

Using fluorescence resonance energy transfer to measure distances along individual DNA molecules: Corrections due to nonideal transfer

Chandran Sabanayagam — 2008-04-21T14:55:34-00:00

The Journal of Chemical Physics, Vol. 122, No. 6. (2005)

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Optimized fabrication and electrical analysis of silver nanowires templated on DNA molecules

Sung Park — 2007-09-28T10:46:22-00:00

Applied Physics Letters, Vol. 89, No. 3. (2006)

We report on the electrical conductivity measurement of silver nanowires templated on native -bacteriophage and synthetic double-stranded DNA molecules. After an electroless chemical deposition, the metallized DNA wires have a diameter down to 15 nm and are among the thinnest metallic nanowires available to date. Two-terminal I-V measurements demonstrating various conduction behaviors are presented. DNA templated functional nanowires may, in the near future, be targeted to connect at specific locations on larger-scale circuits and represent a potential breakthrough in the self-assembly of nanometer-scale structures for electronics layout. ©2006 American Institute of Physics

Antiparallel DNA Double Crossover Molecules As Components for Nanoconstruction

X Li — 2008-05-12T22:22:35-00:00

J. Am. Chem. Soc., Vol. 118, No. 26. (3 July 1996), pp. 6131-6140.

Abstract: Double crossover molecules are DNA structures containing two Holliday junctions connected by two double helical arms. There are several types of double crossover molecules, differentiated by the relative orientations of their helix axes, parallel or antiparallel, and by the number of double helical half-turns (even or odd) between the two crossovers. We have examined these molecules from the viewpoint of their potential utility in nanoconstruction. Whereas the parallel double helical molecules are usually not well behaved, we have focused on the antiparallel molecules; antiparallel molecules with an even number of half turns between crossovers (termed DAE molecules) produce a reporter strand when ligated, so these have been characterized in a ligation cyclization assay. In contrast to other molecules that contain branched junctions, we find that these molecules cyclize rarely or not at all. The double crossover molecules cyclize no more readily than the linear molecule containing the same sequence as the ligation domain. We have tested both a conventional DAE molecule and one containing a bulged three-arm branched junction between the crossovers. The conventional DAE molecule appears to be slightly stiffer, but so few cyclic products are obtained in either case that quantitative comparisons are not possible. Thus, it appears that these molecules may be able to serve as the rigid components that are needed to assemble symmetric molecular structures, such as periodic lattices. We suggest that they be combined with DNA triangles and deltahedra in order to accomplish this goal.

Self-assembly at all scales.

GM Whitesides — 2005-12-17T14:15:26-00:00

Science, Vol. 295, No. 5564. (29 March 2002), pp. 2418-2421.

Self-assembly is the autonomous organization of components into patterns or structures without human intervention. Self-assembling processes are common throughout nature and technology. They involve components from the molecular (crystals) to the planetary (weather systems) scale and many different kinds of interactions. The concept of self-assembly is used increasingly in many disciplines, with a different flavor and emphasis in each.

Molecular self-assembly and nanochemistry: a chemical strategy for the synthesis of nanostructures

GM Whitesides — 2008-02-21T22:19:58-00:00

Science, Vol. 254, No. 5036. (29 November 1991), pp. 1312-1319.

Molecular self-assembly is the spontaneous association of molecules under equilibrium conditions into stable, structurally well-defined aggregates joined by noncovalent bonds. Molecular self-assembly is ubiquitous in biological systems and underlies the formation of a wide variety of complex biological structures. Understanding self-assembly and the associated noncovalent interactions that connect complementary interacting molecular surfaces in biological aggregates is a central concern in structural biochemistry. Self-assembly is also emerging as a new strategy in chemical synthesis, with the potential of generating nonbiological structures with dimensions of 1 to 10(2) nanometers (with molecular weights of 10(4) to 10(10) daltons). Structures in the upper part of this range of sizes are presently inaccessible through chemical synthesis, and the ability to prepare them would open a route to structures comparable in size (and perhaps complementary in function) to those that can be prepared by microlithography and other techniques of microfabrication. 10.1126/science.1962191

An autonomous polymerization motor powered by DNA hybridization

Suvir Venkataraman — 2008-03-19T23:43:07-00:00

Nat Nano, Vol. 2, No. 8. (2007), pp. 490-494.

Challenges and Applications for Self-Assembled DNA Nanostructures

John Reif — 2007-05-07T22:04:55-00:00

(2001), pp. 173-198.

Electronic properties, hydrogen bonding, stacking, and cation binding of DNA and RNA bases.

J Sponer — 2007-09-07T10:38:16-00:00

Biopolymers, Vol. 61, No. 1. (2002), pp. 3-31.

This review summarizes results concerning molecular interactions of nucleic acid bases as revealed by advanced ab initio quantum chemical (QM) calculations published in last few years. We first explain advantages and limitations of modern QM calculations of nucleobases and provide a brief history of this still rather new field. Then we provide an overview of key electronic properties of standard and selected modified nucleobases, such as their charge distributions, dipole moments, polarizabilities, proton affinities, tautomeric equilibria, and amino group hybridization. Then we continue with hydrogen bonding of nucleobases, by analyzing energetics of standard base pairs, mismatched base pairs, thio-base pairs, and others. After this, the nature of aromatic stacking interactions is explained. Also, nonclassical interactions in nucleic acids such as interstrand bifurcated hydrogen bonds, interstrand close amino group contacts, C [bond] H...O interbase contacts, sugar-base stacking, intrinsically nonplanar base pairs, out-of-plane hydrogen bonds, and amino-acceptor interactions are commented on. Finally, we overview recent calculations on interactions between nucleic acid bases and metal cations. These studies deal with effects of cation binding on the strength of base pairs, analysis of specific differences among cations, such as the difference between zinc and magnesium, the influence of metalation on protonation and tautomeric equlibria of bases, and cation-pi interactions involving nucleobases. In this review, we do not provide methodological details, as these can be found in our preceding reviews. The interrelation between advanced QM approaches and classical molecular dynamics simulations is briefly discussed.

Covalent immobilization of DNA onto functionalized mica for atomic force microscopy.

M Ji — 2007-12-12T15:47:31-00:00

J Nanosci Nanotechnol, Vol. 4, No. 6. (July 2004), pp. 580-584.

The immobilization of DNA on the self-assembled monolayer of 3-aminopropyltrimethoxysilane (APTES) on mica wafer functionalized with glutaraldehyde (GA) by chemical bonding was studied by atomic force microscopy (AFM). The DNA used for our investigation was amplified by polymerase chain reaction, and primers were labeled with a -NH2 group at their 5' terminus. The surfaces were analyzed by X-ray photoelectron spectroscopy (XPS) and AFM. Results from XPS and AFM showed that the mica with the APTES and activated with GA can be formed, and the flatness of the mica can be adapted for AFM images. We found that the modified surface was capable of binding DNA molecules so that it withstood a thorough rinsing with a solution of sodium dodecylsulfate. Covalent binding between the aldehyde-terminated membrane and -NH2 groups at both ends of double-stranded DNA resulted in immobilization and straightening of the DNA.

Mastering the complexity of DNA nanostructures

Marco Brucale — 2007-11-26T16:18:06-00:00

Trends in Biotechnology, Vol. 24, No. 5. (May 2006), pp. 235-243.

The self-assembly of oligodeoxynucleotides is a versatile and powerful tool for the construction of objects in the nanoscale. The strictly information-driven pairing of DNA fragments can be used to rationally design and build nanostructures with planned topologies and geometries. Taking advantage of the steadily expanding library of well-characterized DNA motifs, several examples of structures with different dimensionalities have appeared in the literature in the past few years, laying the foundations for a promising DNA-mediated, bottom-up approach to nanotechnology. This article focuses on recent developments in this area of research and proposes a classification of DNA nanostructures based on topological considerations in addition to describing strategies for tackling the inherent complexities of such an endeavor.

Site-directed modification of DNA duplexes by chemical ligation.

NG Dolinnaya — 2008-05-12T14:40:45-00:00

Nucleic acids research, Vol. 16, No. 9. (11 May 1988), pp. 3721-3738.

The efficiency of chemical ligation method have been demonstrated by assembling a number of DNA duplexes with modified sugar phosphate backbone. Condensation on a tetradecanucleotide template of hexa(penta)- and undecanucleotides differing only in the terminal nucleoside residue have been performed using water-soluble carbodiimide as a condensing agent. As was shown by comparing the efficiency of chemical ligation of single-strand breaks in those duplexes, the reaction rate rises 70 or 45 times if the 3'-OH group is substituted with an amino or phosphate group (the yield of products with a phosphoramidate or pyrophosphate bond is 96-100% in 6 d). Changes in the conformation of reacting groups caused by mismatched base pairs (A.A, A.C) as well as the hybrid rU.dA pair or an unpaired base make the template-directed condensation less effective. The thermal stability of DNA duplexes was assayed before and after the chemical ligation. Among all of the modified duplexes, only the duplex containing 3'-rU in the nick was found to be a substrate of T4 DNA ligase.

Remote electronic control of DNA hybridization through inductive coupling to an attached metal nanocrystal antenna

Kimberly Hamad-Schifferli — 2007-09-28T13:27:56-00:00

Nature, Vol. 415, No. 6868. (10 January 2002), pp. 152-155.

Increasingly detailed structural and dynamic studies are highlighting the precision with which biomolecules execute often complex tasks at the molecular scale. The efficiency and versatility of these processes have inspired many attempts to mimic or harness them. To date, biomolecules have been used to perform computational operations and actuation, to construct artificial transcriptional loops that behave like simple circuit elements and to direct the assembly of nanocrystals. Further development of these approaches requires new tools for the physical and chemical manipulation of biological systems. Biomolecular activity has been triggered optically through the use of chromophores, but direct electronic control over biomolecular 'machinery' in a specific and fully reversible manner has not yet been achieved. Here we demonstrate remote electronic control over the hybridization behaviour of DNA molecules, by inductive coupling of a radio-frequency magnetic field to a metal nanocrystal covalently linked to DNA. Inductive coupling to the nanocrystal increases the local temperature of the bound DNA, thereby inducing denaturation while leaving surrounding molecules relatively unaffected. Moreover, because dissolved biomolecules dissipate heat in less than 50 picoseconds (ref. 16), the switching is fully reversible. Inductive heating of macroscopic samples is widely used, but the present approach should allow extension of this concept to the control of hybridization and thus of a broad range of biological functions on the molecular scale.

Nucleic acid junctions and lattices

Nadrian Seeman — 2008-02-21T17:01:20-00:00

Journal of Theoretical Biology, Vol. 99, No. 2. (21 November 1982), pp. 237-247.

It is possible to generate sequences of oligomeric nucleic acids which will preferentially associate to form migrationally immobile junctions, rather than linear duplexes, as they usually do. These structures are predicated on the maximization of Watson-Crick base pairing and the lack of sequence symmetry customarily found in their analogs in living systems. Criteria are presented which oligonucleotide sequences must fulfill in order to yield these junction structures. The generable junctions are nexi, from which 3 to 8 double helices may emanate. Each junction may be treated as a macromolecular "valence cluster", and the individual clusters may be linked together directly, or with pieces of linear DNA interspersed between them. This covalent linkage can be done with enormous specificity, using the sticky-ended ligation techniques currently employed in genetic engineering studies. It appears to be possible to generate covalently joined three-dimensional networks of nucleic acids which are periodic in connectivity and perhaps in space.

Architecture with GIDEON, a program for design in structural DNA nanotechnology.

JJ Birac — 2007-10-22T12:57:40-00:00

J Mol Graph Model, Vol. 25, No. 4. (December 2006), pp. 470-480.

We present geometry based design strategies for DNA nanostructures. The strategies have been implemented with GIDEON-a graphical integrated development environment for oligonucleotides. GIDEON has a highly flexible graphical user interface that facilitates the development of simple yet precise models, and the evaluation of strains therein. Models are built on a simple model of undistorted B-DNA double-helical domains. Simple point and click manipulations of the model allow the minimization of strain in the phosphate-backbone linkages between these domains and the identification of any steric clashes that might occur as a result. Detailed analysis of 3D triangles yields clear predictions of the strains associated with triangles of different sizes. We have carried out experiments that confirm that 3D triangles form well only when their geometrical strain is less than 4% deviation from the estimated relaxed structure. Thus geometry-based techniques alone, without detailed energetic considerations, can be used to explain certain general trends in DNA structure formation. We have used GIDEON to build detailed models of double crossover and triple crossover molecules, evaluating the non-planarity associated with base tilt and junction misalignments. Computer modeling using a graphical user interface overcomes the limited precision of physical models for larger systems, and the limited interaction rate associated with earlier, command-line driven software.

Atomic force microscopy imaging of DNA covalently immobilized on a functionalized mica substrate.

LS Shlyakhtenko — 2007-12-12T15:45:26-00:00

Biophys J, Vol. 77, No. 1. (July 1999), pp. 568-576.

A procedure for covalent binding of DNA to a functionalized mica substrate is described. The approach is based on photochemical cross-linking of DNA to immobilized psoralen derivatives. A tetrafluorphenyl (TFP) ester of trimethyl psoralen (trioxalen) was synthesized, and the procedure to immobilize it onto a functionalized aminopropyl mica surface (AP-mica) was developed. DNA molecules were cross-linked to trioxalen moieties by UV irradiation of complexes. The steps of the sample preparation procedure were analyzed with x-ray photoelectron spectroscopy (XPS). Results from XPS show that an AP-mica surface can be formed by vapor phase deposition of silane and that this surface can be derivatized with trioxalen. The derivatized surface is capable of binding of DNA molecules such that, after UV cross-linking, they withstand a thorough rinsing with SDS. Observations with atomic force microscopy showed that derivatized surfaces remain smooth, so DNA molecules are easily visualized. Linear and circular DNA molecules were photochemically immobilized on the surface. The molecules are distributed over the surface uniformly, indicating rather even modification of AP-mica with trioxalen. Generally, the shapes of supercoiled molecules electrostatically immobilized on AP-mica and those photocross-linked on trioxalen-functionalized surfaces remain quite similar. This suggests that UV cross-linking does not induce formation of a noticeable number of single-stranded breaks in DNA molecules.

Scanning Force Microscopy of DNA Deposited onto Mica: EquilibrationversusKinetic Trapping Studied by Statistical Polymer Chain Analysis

Claudio Rivetti — 2007-11-22T18:06:06-00:00

Journal of Molecular Biology, Vol. 264, No. 5. (20 December 1996), pp. 919-932.

This paper reports a study of the deposition process of DNA molecules onto a mica surface for imaging under the scanning force microscope (SFM). Kinetic experiments indicate that the transport of DNA molecules from the solution drop onto the surface is governed solely by diffusion, and that the molecules are irreversibly adsorbed onto the substrate. A statistical polymer chain analysis has been applied to DNA molecules to determine the deposition conditions that lead to equilibrium and those that result in trapped configurations. Using the appropriate conditions, DNA molecules deposited onto freshly cleaved mica, are able to equilibrate on the surface as in an ideal two-dimensional solution. A persistence length of 53 nm was determined from those molecules. DNA fragments that were labeled on both ends with a horseradish peroxidase streptavidin fusion protein were still able to equilibrate on the surface, despite the additional protein-surface interaction. In contrast, DNA molecules deposited onto glow-discharged mica or H+-exchanged mica do not equilibrate on the surface. These molecules adopt conformations similar to those expected for a simple projection onto the surface plane, suggesting a process of kinetic trapping. These results validate recent SFM application to quantitatively analyze the conformation of complex macromolecular assemblies deposited on mica. Under equilibration conditions, the present study indicates that the SFM can be used to determine the persistence length of DNA molecules to a high degree of precision.

Algorithmic self-assembly of DNA Sierpinski triangles.

PW Rothemund — 2005-05-02T18:13:02-00:00

PLoS Biol, Vol. 2, No. 12. (December 2004)

Algorithms and information, fundamental to technological and biological organization, are also an essential aspect of many elementary physical phenomena, such as molecular self-assembly. Here we report the molecular realization, using two-dimensional self-assembly of DNA tiles, of a cellular automaton whose update rule computes the binary function XOR and thus fabricates a fractal pattern--a Sierpinski triangle--as it grows. To achieve this, abstract tiles were translated into DNA tiles based on double-crossover motifs. Serving as input for the computation, long single-stranded DNA molecules were used to nucleate growth of tiles into algorithmic crystals. For both of two independent molecular realizations, atomic force microscopy revealed recognizable Sierpinski triangles containing 100-200 correct tiles. Error rates during assembly appear to range from 1% to 10%. Although imperfect, the growth of Sierpinski triangles demonstrates all the necessary mechanisms for the molecular implementation of arbitrary cellular automata. This shows that engineered DNA self-assembly can be treated as a Turing-universal biomolecular system, capable of implementing any desired algorithm for computation or construction tasks.

Programmable DNA self-assemblies for nanoscale organization of ligands and proteins.

SH Park — 2006-02-09T18:35:17-00:00

Nano Lett, Vol. 5, No. 4. (April 2005), pp. 729-733.

We demonstrate the precise control of periodic spacing between individual protein molecules by programming the self-assembly of DNA tile templates. In particular, we report the application of two self-assembled periodic DNA structures, two-dimensional nanogrids, and one-dimensional nanotrack, as template for programmable self-assembly of streptavidin protein arrays with controlled density.

DNA-scaffolded nanoparticle structures

Björn Högberg — 2007-04-20T12:38:58-00:00

J. Phys.: Conf. Ser., Vol. 61, No. 1. (2007), 458.