Electrochemical DNA detection systems are an attractive approach to the development of multiplexed, high-throughput DNA analysis systems for clinical and research applications. We have engineered a new class of nanoelectrode ensembles (NEEs) that constitute a useful platform for biomolecular electrochemical sensing. High-sensitivity DNA detection was achieved at oligonucleotide-functionalized NEEs using a label-free electrocatalytic assay. Attomole levels of DNA were detected using the NEEs, validating the promise of nanoarchitectures for ultrasensitive biosensing.

Integrating nanotechnology with experimental biology is paramount to advancing fundamental biological science and technology, and, therefore, of high current interest and importance. In this article, we report on a new possibility of utilizing carbon nanotube probes assembled by a modified dielectrophoretic based technique for single-cell experimentation and delivery. The modified approach permits highly reproducible construction of water-stable, highly-aligned, and electrically-conductive probes several hundred microns in length, which hold a great promise for enhancing previously developed molecular-scale intracellular experimental techniques. The results of this work, in particular, indicate that the minimally invasive nanotube probes could be advantageous for studies involving permeabilization and subsequent desorption of molecules into a cell's interior, thereby obviating permeabilization and diffusion across membranes.

We describe a strategy that permits discrete regions of arrayed carbon nanotubes (CNTs) to be functionalized simultaneously and specifically with DNA oligonucleotides. The different chemical properties of two regions on single CNTs and orthogonal chemical coupling strategies have been exploited to derivatize CNTs within highly ordered arrays with multiple DNA sequences. Through duplex hybridization, we then targeted different DNA sequences with appended metal nanoparticles to distinct sites on the CNT architecture with precise spatial control. The materials generated from these studies represent the first CNTs with bipartite functionalization. The approach described provides a high level of precision in parallel and directed assembly of DNA sequences and appended cargo and is useful for the preparation of novel hybrid bionanomaterials.

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As part of an effort to develop nanoelectronic sensors for biological targets, we tested the potential to incorporate coiled coils as metallized, self-assembling, site-specific molecular linkers on carbon nanotubes (CNTs). Based on a previously conceived modular anchor-probe approach, a system was designed in which hydrophobic residues (valines and leucines) form the interface between the two helical peptide components. Charged residues (glutamates and arginines) on the borders of the hydrophobic interface increase peptide solubility, and provide stability and specificity for anchor-probe assembly. Two histidine residues oriented on the exposed hydrophilic exterior of each peptide were included as chelating sites for metal ions such as cobalt. Cysteines were incorporated at the peptide termini for oriented, thiol-mediated coupling to surface plasmon resonance (SPR) biosensor surfaces, gold nanoparticles or CNT substrates. The two peptides were produced by solid phase peptide synthesis using Fmoc chemistry: an acidic 42-residue peptide E42C, and its counterpart in the heterodimer, a basic 39-residue peptide R39C. The ability of E42C and R39C to bind cobalt was demonstrated by immobilized metal affinity chromatography and isothermal titration calorimetry. SPR biosensor kinetic analysis of dimer assembly revealed apparent sub-nanomolar affinities in buffers with and without 1 mM CoCl2 using two different reference surfaces. For device-oriented CNT immobilization, R39C was covalently anchored to CNT tips via a C-terminal cysteine residue. Scanning electron microscopy was used to visualize the assembly of probe peptide (E42C) N-terminally labeled with 15 nm gold nanoparticles, when added to the R39C-CNT surface. The results obtained open the way to develop CNT tip-directed recognition surfaces, using recombinant and chemically synthesized chimeras containing binding epitopes fused to the E42C sequence domain.

A strategy of metallizing peptides to serve as conduits of electronic signals that bridge between a redox enzyme and a carbon-nanotube electrode has been developed with enhanced results. In conjunction, a protocol to link the biological elements to the tips of carbon nanotubes has been developed to optimize contact and geometry between the redox enzyme and the carbon nanotube electrode array. A peptide nanowire of 33 amino acids, comprised of a leucine zipper motif, was mutated to bind divalent metals, conferring conductivity into the peptide. Reaction between a thiolate of the peptide with the sulfenic acid of the NADH peroxidase enzyme formed a peptide-enzyme assembly that are fully primed to transduce electrons out of the enzyme active site to an electrode. Scanning electron microscopy shows immobilization and linking of the assembly specifically to the tips of carbon nanotube electrodes, as designed. Isothermal titration calorimetry and mass spectrometry indicate a binding stoichiometry of at least three metals bound per peptide strand. Overall, these results highlight the gain that can be achieved when the signal tranducing units of a biosensor are aligned through directed peptide chemistry.

This paper describes a class of three component hybrid nanowires templated by DNA directed self-assembly. Through the modification of carbon nanotube (CNT) termini with synthetic DNA oligonucleotides, gold nanoparticles are delivered, via DNA hybridization, to CNT tips that then serve as growth sites for zinc oxide (ZnO) nanowires. The structures we have generated using DNA templating represent an advance toward building higher order sequenced one dimensional nanostructures with rational control.

The authors describe the properties of electronically active nanowires that can be assembled via DNA directed growth on a nanostructured array. DNA-modified nanoparticles are used to site-specifically address the tips of vertically aligned carbon nanotubes (CNTs) that serve as catalysts for the growth of zinc oxide (ZnO) nanorods. Using conductive probe atomic force microscopy, they measured the conductance characteristics of single ZnO-CNT structures under various force and illumination conditions and at different sites in a large array, thereby establishing that DNA directed formation of multimaterial, optically active nanostructures can yield devices that are electronically functional at the nanometer scale. The inherent ability of DNA to carry and convey encoded information provides the basis for targeted synthesis of nanostructured devices.