Bacteria-derived topologies of Cu2O nanozymes exert a variable antibacterial effect.

The ability of bacteria to facilitate fabrication of nanomaterials has been adapted towards bacterial sensing applications. In this work, we fabricate spherical, cubic and truncated octahedron topologies of Cu2O nanoparticles via E. coli-facilitated redox reaction in an electrochemical setup. The Cu2O nanoparticles exhibit cytochrome c oxidase-like activity with the spherical topology displaying higher catalytic rate compared to the other geometries. The topology-dependent catalytic behavior of Cu2O nanoparticles has not been reported previously. The Cu2O nanozymes also display E. coli killing activity in a topology-correlated manner. The E. coli mediated redox reaction in an electrochemical setup is being reported for the first time for synthesis of different topologies of Cu2O which also exert a variable antibacterial effect.

pH-Responsive and Reversible A-Motif-Based DNA Hydrogel: Synthesis and Biosensing Application.

Functional DNA hydrogels with various motifs and functional groups require perfect sequence design to avoid cross-bonding interference with themselves or other structural sequences. This work reports an A-motif functional DNA hydrogel that does not require any sequence design. A-motif DNA is a noncanonical parallel DNA duplex structure containing homopolymeric deoxyadenosines (poly-dA) strands that undergo conformation changes from single strands at neutral pH to a parallel duplex DNA helix at acidic pH. Despite this and other advantages over other DNA motifs like no cross-bonding interference with other structural sequences, the A-motif has not been explored much. We successfully synthesized a DNA hydrogel by using an A-motif as a reversible handle to polymerize a DNA three-way junction. The A-motif hydrogel was initially characterized by electrophoretic mobility shift assay, and dynamic light scattering, which showed the formation of higher-order structures. Further, we used imaging techniques like atomic force microscopy and scanning electron microscope to validating its hydrogel like highly branched morphology. pH-induced conformation transformation from monomers to gel is quick and reversible, and was analysed for multiple acid-base cycles. The sol-to-gel transitions and gelation properties were further examined in rheological studies. The use of the A-motif hydrogel in the visual detection of pathogenic target nucleic acid sequence was demonstrated for the first time in a capillary assay. Moreover, pH-induced hydrogel formation was observed in situ as a layer over the mammalian cells. The proposed A-motif DNA scaffold has enormous potential in designing stimuli-responsive nanostructures that can be used for many biological applications.

Geometry of a DNA Nanostructure Influences Its Endocytosis: Cellular Study on 2D, 3D, and in Vivo Systems.

Fabrication of nanoscale DNA devices to generate 3D nano-objects with precise control of shape, size, and presentation of ligands has shown tremendous potential for therapeutic applications. The interactions between the cell membrane and different topologies of 3D DNA nanostructures are crucial for designing efficient tools for interfacing DNA devices with biological systems. The practical applications of these DNA nanocages are still limited in cellular and biological systems owing to the limited understanding of their interaction with the cell membrane and endocytic pathway. The correlation between the geometry of DNA nanostructures and their internalization efficiency remains elusive. We investigated the influence of the shape and size of 3D DNA nanostructures on their cellular internalization efficiency. We found that one particular geometry, i.e., the tetrahedral shape, is more favored over other designed geometries for their cellular uptake in 2D and 3D cell models. This is also replicable for cellular processes like cell invasion assays in a 3D spheroid model, and passing the epithelial barriers in in vivo zebrafish model systems. Our work provides detailed information for the rational design of DNA nanodevices for their upcoming biological and biomedical applications.

Designer DNA Hydrogels Stimulate 3D Cell Invasion by Enhanced Receptor Expression and Membrane Endocytosis.

DNA has emerged as one of the smartest biopolymers to bridge the gap between chemical science and biology to design scaffolds like hydrogels by physical entanglement or chemical bonding with remarkable properties. We present here a completely new application of DNA-based hydrogels in terms of their capacity to stimulate membrane endocytosis, leading to enhanced cell spreading and invasion for cells in ex vivo 3D spheroids models. Multiscale simulation studies along with DLS data showed that the hydrogel formation was enhanced at lower temperature and it converts to liquid with increase in temperature. DNA hydrogels induced cell spreading as observed by the increase in cellular area by almost two-fold followed by an increase in the receptor expression, the endocytosis, and the 3D invasion potential of migrating cells. Our first results lay the foundation for upcoming diverse applications of hydrogels to probe and program various cellular and physiological processes that can have lasting applications in stem cell programming and regenerative therapeutics.

Stimuli Responsive, Programmable DNA Nanodevices for Biomedical Applications.

Of the multiple areas of applications of DNA nanotechnology, stimuli-responsive nanodevices have emerged as an elite branch of research owing to the advantages of molecular programmability of DNA structures and stimuli-responsiveness of motifs and DNA itself. These classes of devices present multiples areas to explore for basic and applied science using dynamic DNA nanotechnology. Herein, we take the stake in the recent progress of this fast-growing sub-area of DNA nanotechnology. We discuss different stimuli, motifs, scaffolds, and mechanisms of stimuli-responsive behaviours of DNA nanodevices with appropriate examples. Similarly, we present a multitude of biological applications that have been explored using DNA nanodevices, such as biosensing, in vivo pH-mapping, drug delivery, and therapy. We conclude by discussing the challenges and opportunities as well as future prospects of this emerging research area within DNA nanotechnology.

DNA-Functionalized Nanoparticles for Targeted Biosensing and Biological Applications.

Nanoscale systems have increasingly been used in biomedical applications, enhancing the demand for the development of biomolecule-functionalized nanoparticles for targeted applications. Such designer nanosystems hold great prospective to refine disease diagnosis and treatment. To completely investigate their potential for bioapplications, nanoparticles must be biocompatible and targetable toward explicit receptors to guarantee particular detecting, imaging, and medication conveyance in complex organic milieus, for example, living cells, tissues, and organisms. We present recent works that explore enhanced biocompatibility and biorecognition of nanoparticles functionalized with DNA and different DNA entities such as aptamers, DNAzymes, and aptazymes. We sum up the methods utilized in the amalgamation of complex nanostructures, survey the significant types of multifunctional nanoparticles that have been developed in the course of recent years, and give a perceptual vision of the significant field of nanomedicine. The field of DNA-functionalized nanoparticles holds an incredible guarantee in rising biomedical zones, for example, multimodal imaging, theranostics, and picture-guided treatments.

DNA Nanodevices to Probe and Program Membrane Organization, Dynamics, and Applications.

Continuous, dynamic, and controlled membrane remodeling creates flow of information and materials across membranes to sustain life in all biological systems. Multiple nanoscale phenomena of membranes regulate mesoscale processes in cells, which in turn control macro-scale processes in living organisms. Understanding the molecular mechanisms that cells use for membrane homeostasis, i.e., to generate, maintain, and deform the membrane structures has therefore been the mammoth's task in biology. Using the principles of DNA nanotechnology, researchers can now precisely recapitulate the functional interactions of the biomolecules that can now probe, program, and re-program membrane remodeling and associated phenomena. The molecular mechanisms for membrane dynamics developing in vitro conditions in which the membrane modulating components are precisely organized and modulated by DNA nanoscaffolds are adding new chapters in the field of DNA nanotechnology. In this review, we discuss DNA nanodevices-based membrane remodeling and trafficking machineries and their applications in biological systems.

Functional DNA Based Hydrogels: Development, Properties and Biological Applications.

DNA-based nanostructures have emerged as a versatile component for nanoscale construction of soft materials. Multiple structural, functional properties and versatility in conjugation with other biomolecules made DNA the material of choice to use in various biomedical applications. DNA-based hydrogels significantly attracted attention in recent years owing to their properties and applications in biosensing, bioimaging, and therapeutics. Here, we summarize the recent advances in the area of DNA hydrogels where these are used either as structural material or as functional entities to make hybrid constructs with various biomedical applications. Multiple synthetic routes for constructing DNA hydrogels are summarized first, where the structural motifs and spatial arrangements are considered for the classification of DNA materials. We then present the characterization and properties of DNA hydrogels using multiple imaging and biophysical techniques. Further, different biomedical applications of DNA hydrogels are presented such as biosensing, bioimaging, and targeted drug delivery and as scaffolds to program cellular systems. Last, we discuss the vision and potential of DNA based hydrogels as an emerging class of therapeutically important devices for theragnostic and other biological applications.

Nanobiocatalyst facilitated aglycosidic quercetin as a potent inhibitor of tau protein aggregation.

Polyphenols have been suggested as potential therapeutic agents for the treatment of amyloidogenic diseases. In this work, we evaluate quercetin-rich onion extract for its ability to inhibit tau fibrillization. Considering the presence of polyphenols in multiple glycosidic and aglycosidic forms, a nanobiocatalyst-mediated approach has been used to extract quercetin from onion skins. The nanobiocatalysts facilitate greater release of quercetin compared to the use of free enzymes. Atomic force microscopy and fluorescence microscopy show that quercetin possesses a novel inhibitory character on tau-fibril aggregation. In contrast, quercetin-diglucoside does not have an inhibitory effect. Molecular Dynamics simulations reveal conformational changes in tau protein upon interaction with quercetin due to specific hydrogen bonding and hydrophobic interactions. The resulting conformational stability of tau monomer reduces propensity of the protein to aggregate. The ability of quercetin to inhibit tau fibrillization expands the paradigm for application of bioactive polyphenols.

Investigation of nanoparticle immobilized cellulase: nanoparticle identity, linker length and polyphenol hydrolysis.

Cellulase containing nanobiocatalysts have been useful as an extraction tool based on their ability to disrupt plant cell walls. In this work, we investigate the effect of nanoparticle composition and chemical linkage towards immobilized cellulase activity. Cellulase nanoconstructs have been prepared, characterized and compared for their loading efficiencies with standard assays and enzyme kinetics and correlate well with the cognate loading efficiencies. Application of the cellulase-immobilized nanoparticles on onion skins results in release of a distinctive composition of polyphenols. The aglycosidic form of quercetin is the dominant product of onion skin hydrolysis affected by cellulase nanobiocatalysts. Chitosan-coated iron oxide nanoparticles with APTES-conjugated cellulase are found to be most effective for polyphenol release and for transformation of glycosidic to aglycosidic form of quercetin. These results shed light on the activity of immobilized cellulase beyond their role in cell wall disruption and are important for the practical application of cellulase nanobiocatalysts.