ABSTRACT
The number of applications of self-assembled deoxyribonucleic acid (DNA) origami nanoparticles (DNA NPs) has increased drastically, following the development of a variety of single-stranded template DNA (ssDNA) that can serve as the scaffold strand. In addition to viral genomes, such as M13 bacteriophage and lambda DNAs, enzymatically produced ssDNA from various template sources is rapidly gaining traction and being applied as the scaffold for DNA NP preparation. However, separating fully formed DNA NPs that have custom scaffolds from crude assembly mixes is often a multistep process of first separating the ssDNA scaffold from its enzymatic amplification process and then isolating the assembled DNA NPs from excess precursor strands. Only then is the DNA NP sample ready for downstream characterization and application. In this work, we highlight a single-step purification of custom sequence- or M13-derived scaffold-based DNA NPs using photocleavable biotin tethers. The process only requires an inexpensive ultraviolet (UV) lamp, and DNA NPs with up to 90% yield and high purity are obtained. We show the versatility of the process in separating two multihelix bundle structures and a wireframe polyhedral architecture.
Subject(s)
Biotin , DNA, Single-Stranded , Nanoparticles , Biotin/chemistry , Nanoparticles/chemistry , DNA, Single-Stranded/chemistry , DNA, Single-Stranded/isolation & purification , Bacteriophage M13/chemistry , Bacteriophage M13/genetics , DNA/chemistry , DNA/isolation & purification , Ultraviolet RaysABSTRACT
The formation of beta-amyloid (Aß) plaques is a classical hallmark of Alzheimer's disease (AD) that is associated with the promotion of neuroinflammation and subsequent neurotoxicity. Given the limited therapeutic options for targeting and clearing Aß plaques in AD, there is an urgent need to develop effective approaches to reduce plaque accumulation. The objective of this study was to validate mild magnetic nanoparticle (MNP) hyperthermia technology as a strategy to clear Aß deposits and determine the impact on microglia functionality. Our results demonstrated that the heating of MNPs localized to Aß aggregates upon exposure to high frequency alternating magnetic field (AMF) was sufficient to disrupt Aß plaques, resulting in its fragmentation. Importantly, this could facilitate the phagocytic clearance of Aß as well as attenuate pro-inflammatory responses by human microglial cells. Our results support the feasibility of mild MNP/AMF hyperthermia as a new strategy for reducing beta-amyloid burdens in Alzheimer's disease.
Subject(s)
Amyloid beta-Peptides/isolation & purification , Hyperthermia, Induced/methods , Magnetics , Microglia/metabolism , Nanoparticles , Plaque, Amyloid/metabolism , Amyloid beta-Peptides/metabolism , Cell Line, Transformed , Humans , Microscopy, Electron, Scanning , PhagocytosisABSTRACT
Currently, DNA nanotechnology offers the most programmable, scalable, and accurate route for the self-assembly of matter with nanometer precision into 1, 2, or 3D structures. One example is DNA origami that is well suited to serve as a molecularly defined "breadboard", and thus, to organize various nanomaterials such as nanoparticles into hybrid systems. Since the controlled assembly of quantum dots (QDs) is of high interest in the field of photonics and other optoelectronic applications, a more detailed view on the functionalization of QDs with oligonucleotides shall be achieved. In this work, four different methods are presented to characterize the functionalization of thiol-capped cadmium telluride QDs with oligonucleotides and for the precise quantification of the number of oligonucleotides bound to the QD surface. This study enables applications requiring the self-assembly of semiconductor-oligonucleotide hybrid materials and proves the conjugation success in a simple and straightforward manner.
Subject(s)
Nanotechnology/methods , Oligonucleotides/chemistry , Quantum Dots/chemistry , Cadmium Compounds/chemistry , DNA/chemistry , Edetic Acid/chemistry , Fluorescent Dyes/chemistry , Luminescence , Spectrum Analysis , Tellurium/chemistryABSTRACT
The synthesis, purification, and structure characterization of a seven-ring interlocked DNA catenane is described. The design of the seven-ring catenane allows the dynamic reconfiguration of any of the four rings (R1, R3, R4, and R6) on the catenane scaffold, or the simultaneous switching of any combination of two, three, or all four rings to yield 16 different isomeric states of the catenane. The dynamic reconfiguration across the states is achieved by implementing the strand-displacement process in the presence of appropriate fuel/antifuel strands and is probed by fluorescence spectroscopy. Each of the 16 isomers of the catenane can be transformed into any of the other isomers, thus allowing for 240 dynamic transitions within the system.
