Conjugation with artificial nucleic acids allows proteins to be modified with a synthetically accessible, robust tag. This attachment is addressable in a highly specific manner by means of molecular ...recognition events, such as Watson-Crick hybridization. Such DNA-protein conjugates, with their combined properties, have a broad range of applications, such as in high-performance biomedical diagnostic assays, fundamental research on molecular recognition, and the synthesis of DNA nanostructures. This Review surveys current approaches to generate DNA-protein conjugates as well as recent advances in their applications. For example, DNA-protein conjugates have been assembled into model systems for the investigation of catalytic cascade reactions and light-harvesting devices. Such hybrid conjugates are also used for the biofunctionalization of planar surfaces for micro- and nanoarrays, and for decorating inorganic nanoparticles to enable applications in sensing, materials science, and catalysis.
In the past 35 years, DNA nanotechnology has grown to a highly innovative and vibrant field of research at the interface of chemistry, materials science, biotechnology, and nanotechnology. Herein, a ...short summary of the state of research in various subdisciplines of DNA nanotechnology, ranging from pure “structural DNA nanotechnology” over protein–DNA assemblies, nanoparticle‐based DNA materials, and DNA polymers to DNA surface technology is given. The survey shows that these subdisciplines are growing ever closer together and suggests that this integration is essential in order to initiate the next phase of development. With the increasing implementation of machine‐based approaches in microfluidics, robotics, and data‐driven science, DNA‐material systems will emerge that could be suitable for applications in sensor technology, photonics, as interfaces between technical systems and living organisms, or for biomimetic fabrication processes.
DNA nanotechnology is on its way to creating integrated functional systems for material research, life sciences, and high‐end technology. The current state of various subdisciplines of DNA nanotechnology is summarized, ranging from “structural DNA nanotechnology” over protein‐ and nanoparticle‐modified DNA assemblies, to DNA polymers and DNA surface technology, respectively.
DNA Origami: The Art of Folding DNA Saccà, Barbara; Niemeyer, Christof M.
Angewandte Chemie (International ed.),
January 2, 2012, Letnik:
51, Številka:
1
Journal Article
Recenzirano
The advent of DNA origami technology greatly simplified the design and construction of nanometer‐sized DNA objects. The self‐assembly of a DNA‐origami structure is a straightforward process in which ...a long single‐stranded scaffold (often from the phage M13mp18) is folded into basically any desired shape with the help of a multitude of short helper strands. This approach enables the ready generation of objects with an addressable surface area of a few thousand nm2 and with a single “pixel” resolution of about 6 nm. The process is rapid, puts low demands on experimental conditions, and delivers target products in high yields. These features make DNA origami the method of choice in structural DNA nanotechnology when two‐ and three‐dimensional objects are desired. This Minireview summarizes recent advances in the design of DNA origami nanostructures, which open the door to numerous exciting applications.
Know when to fold ′em: As in the ancient art of paper folding, where a single sheet of paper is modeled into beautiful shapes, DNA origami technology allows nanoscale‐addressable objects to be created from one single strand of DNA (see picture).
Biological compartmentalization is a fundamental principle of life that allows cells to metabolize, propagate, or communicate with their environment. Much research is devoted to understanding this ...basic principle and to harness biomimetic compartments and catalytic cascades as tools for technological processes. This Review summarizes the current state‐of‐the‐art of these developments, with a special emphasis on length scales, mass transport phenomena, and molecular scaffolding approaches, ranging from small cross‐linkers over proteins and nucleic acids to colloids and patterned surfaces. We conclude that the future exploration and exploitation of these complex systems will largely benefit from technical solutions for the integrated, machine‐assisted development and maintenance of a next generation of biotechnological processes. These goals should be achievable by implementing microfluidics, robotics, and added manufacturing techniques supplemented by theoretical simulations as well as computer‐aided process modeling based on big data obtained from multiscale experimental analyses.
Machine‐generated multienzyme cascades: The machine‐assisted development of biomimetic compartments and catalytic cascades will pave the way towards a novel generation of biotechnological processes. Molecular scaffolds with small cross‐linkers, proteins, nucleic acids, colloids, and patterned surfaces can be used to arrange the catalytic units. S: substrate, P: product.
Chemical Strategies for Generating Protein Biochips Jonkheijm, Pascal; Weinrich, Dirk; Schröder, Hendrik ...
Angewandte Chemie (International ed.),
December 1, 2008, Letnik:
47, Številka:
50
Journal Article
Recenzirano
Protein biochips are at the heart of many medical and bioanalytical applications. Increasing interest has been focused on surface activation and subsequent functionalization strategies for ...immobilizing these biomolecules. Different approaches using covalent and noncovalent chemistry are reviewed; particular emphasis is placed on the chemical specificity of protein attachment and on retention of protein function. Strategies for creating protein patterns (as opposed to protein arrays) are also outlined. An outlook on promising and challenging future directions for protein biochip research and applications is also offered.
The positioning of enzymes on DNA nanostructures for the study of spatial effects in interacting biomolecular assemblies requires chemically mild immobilization procedures as well as efficient means ...for separating unbound proteins from the assembled constructs. We herein report the exploitation of free‐flow electrophoresis (FFE) for the purification of DNA origami structures decorated with biotechnologically relevant recombinant enzymes: the S‐selective NADP+/NADPH‐dependent oxidoreductase Gre2 from S. Cerevisiae and the reductase domain of the monooxygenase P450 BM3 from B. megaterium. The enzymes were fused with orthogonal tags to facilitate site‐selective immobilization. FFE purification yielded enzyme–origami constructs whose specific activity was quantitatively analyzed. All origami‐tethered enzymes were significantly more active than the free enzymes, thereby suggesting a protective influence of the large, highly charged DNA nanostructure on the stability of the proteins.
A functional art form: The efficient separation of enzyme‐decorated DNA origami structures from unbound proteins by free‐flow electrophoresis (FFE; see picture) enabled a differentiated assessment of the activity of origami‐tethered and non‐DNA‐bound enzymes. It was found that enzyme arranged on origami structures were more active than the free enzymes.
The natural micro‐ and nanoscale organization of biomacromolecules is a remarkable principle within living cells, allowing for the control of cellular functions by compartmentalization, dimensional ...diffusion and substrate channeling. In order to explore these biological mechanisms and harness their potential for applications such as sensing and catalysis, molecular scaffolding has emerged as a promising approach. In the case of synthetic enzyme cascades, developments in DNA nanotechnology have produced particularly powerful scaffolds whose addressability can be programmed with nanometer precision. In this minireview, we summarize recent developments in the field of biomimetic multicatalytic cascade reactions organized on DNA nanostructures. We emphasize the impact of the underlying design principles like DNA origami, efficient strategies for enzyme immobilization, as well as the importance of experimental design parameters and theoretical modeling. We show how DNA nanostructures have enabled a better understanding of diffusion and compartmentalization effects at the nanometer length scale, and discuss the challenges and future potential for commercial applications.
Nucleic acid nanostructures are capable of precisely arranging proteins at nanoscale distances and have thus proven to be powerful scaffolds for studying mechanistic effects in enzyme cascades. This minireview summarizes the current status of proposed underlying mechanistic effects, derives aspects for further systematic analysis, and discusses future perspectives for biocatalytic applications.
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.
At the crossroads of biotechnology, nanoscience, and materials research lies a new interdisciplinary research area, in which the use of evolutionary optimized biomolecules for the development of advanced materials and analytical procedures is investigated. As an example, nanoparticles can be interconnected with DNA to generate nanostructured hybrid materials (see scanning force microscopy image).
Rational Design of DNA Nanoarchitectures Feldkamp, Udo; Niemeyer, Christof M.
Angewandte Chemie (International ed.),
March 13, 2006, Letnik:
45, Številka:
12
Journal Article
Recenzirano
DNA has many physical and chemical properties that make it a powerful material for molecular constructions at the nanometer length scale. In particular, its ability to form duplexes and other ...secondary structures through predictable nucleotide‐sequence‐directed hybridization allows for the design of programmable structural motifs which can self‐assemble to form large supramolecular arrays, scaffolds, and even mechanical and logical nanodevices. Despite the large variety of structural motifs used as building blocks in the programmed assembly of supramolecular DNA nanoarchitectures, the various modules share underlying principles in terms of the design of their hierarchical configuration and the implemented nucleotide sequences. This Review is intended to provide an overview of this fascinating and rapidly growing field of research from the structural design point of view.
In the past 20 years, DNA has been established as a powerful material for molecular constructions at the nanometer scale. The ability to design building blocks from DNA containing well‐defined secondary structure motifs (a–f, in the scheme) allows the assembly of large supramolecular arrays, scaffolds, and even mechanical and logical nanodevices.
Proteins possess intrinsic functionalities, which have been optimized in billions of years of natural evolution. The conjugation of proteins with artificial nucleic acids allows one to further ...functionalize proteins with a synthetically accessible, physicochemically robust tag, which is addressable in a highly specific manner by Watson-Crick hybridization. The resulting DNA-protein conjugates can be advantageously used in a variety of applications, ranging from biomedical diagnostics to DNA-based nanofabrication. This critical review provides an overview on chemical approaches to the synthesis of DNA-protein conjugates and their applications in biomolecular nanosciences (96 references).
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