Point-of-care nucleic acid tests: assays and devices.

The COVID-19 pandemic (caused by the SARS_CoV_2 virus) has emphasized the need for quick, easy-to-operate, reliable, and affordable diagnostic tests and devices at the Point-of-Care (POC) for homes/fields/clinics. Such tests and devices will contribute significantly to the fight against the COVID-19 pandemic and any future infectious disease epidemic. Often, academic research studies and those from industry lack knowledge of each other's developments.

RNA Nanostructures: From Structure to Function.

Nucleic-acid nanostructures, which have been designed and constructed with atomic precision, have been used as scaffolds for different molecules and proteins, as nanomachines, as computational components, and more. In particular, RNA has garnered tremendous interest as a building block for the self-assembly of sophisticated and functional nanostructures by virtue of its ease of synthesis by or transcription, its superior mechanical and thermodynamic properties, and its functional roles in nature. In this Topical Review, we describe recent developments in the use of RNA for the design and construction of nanostructures.

APOBEC3G enhances lymphoma cell radioresistance by promoting cytidine deaminase-dependent DNA repair.

APOBEC3 proteins catalyze deamination of cytidines in single-stranded DNA (ssDNA), providing innate protection against retroviral replication by inducing deleterious dC > dU hypermutation of replication intermediates. APOBEC3G expression is induced in mitogen-activated lymphocytes; however, no physiologic role related to lymphoid cell proliferation has yet to be determined. Moreover, whether APOBEC3G cytidine deaminase activity transcends to processing cellular genomic DNA is unknown.

DNA nanotechnology.

The base sequence encoded in nucleic acids yields significant structural and functional properties into the biopolymer. The resulting nucleic acid nanostructures provide the basis for the rapidly developing area of DNA nanotechnology. Advances in this field will be exemplified by discussing the following topics: (i) Hemin/G-quadruplex DNA nanostructures exhibit unique electrocatalytic, chemiluminescence and photophysical properties.

Self-assembly of DNA nanotubes with controllable diameters.

The synthesis of DNA nanotubes is an important area in nanobiotechnology. Different methods to assemble DNA nanotubes have been reported, and control over the width of the nanotubes has been achieved by programmed subunits of DNA tiles. Here we report the self-assembly of DNA nanotubes with controllable diameters.

Generation of photocurrents by bis-aniline-cross-linked Pt nanoparticle/photosystem I composites on electrodes.

Pt nanocrystals are implanted into photosystem I (PSI) by a photochemical reaction. The PSI with the associated Pt nanoclusters was modified with thioaniline and electropolymerized with thioaniline-functionalized Pt nanoparticles (NPs) to yield a bis-aniline-cross-linked PSI/Pt NPs composite. The alignment of the PSI with respect to the Pt NPs leads to effective charge separation and to generation of a photocurrent, ϕ (λ = 420 nm) = 2.

Covalently linked DNA nanotubes.

The present study introduces an approach to prepare covalently linked DNA nanotubes. A circular DNA that includes at its opposite poles thiol and amine functionalities acts as the building block for the construction of the DNA nanotubes. The circular DNA is cross-linked with a bis-amide-modified nucleic acid to yield DNA nanowires, and these are subsequently cross-linked by a bis-thiolated nucleic acid to yield the DNA nanotubes.

Self-assembly of aptamer-circular DNA nanostructures for controlled biocatalysis.

Two kinds of circular DNA components are generated by the hybridization of short nucleic acids with the 3' and 5' ends of single-stranded DNA chains. The circular DNA components include, each, complementary domains for the anticocaine aptamer subunits, and sequence-specific domains for the auxiliary hybridization of programmed nucleic acid-functionalized proteins. The circular DNA components are self-assembled, in the presence of cocaine, into DNA nanowires (micrometer-long nanowires exhibiting heights of ca.

Control of bioelectrocatalytic transformations on DNA scaffolds.

The spatial organization of biomolecules on a DNA scaffold linked to an electrode leads to programmed biocatalytic transformations. This is exemplified by the electrical contacting of glucose oxidase (GOx) linked to the DNA scaffold with the electrode. A nucleic acid functionalized with a ferrocene relay unit was hybridized with the DNA scaffold at a position adjacent to the electrode, and GOx functionalized with nucleic acid units complementary to the specific domain of the DNA template was hybridized with the DNA scaffold in a position remote from the electrode.

Enzyme cascades activated on topologically programmed DNA scaffolds.

The ability of DNA to self-assemble into one-, two- and three-dimensional nanostructures, combined with the precision that is now possible when positioning nanoparticles or proteins on DNA scaffolds, provide a promising approach for the self-organization of composite nanostructures. Predicting and controlling the functions that emerge in self-organized biomolecular nanostructures is a major challenge in systems biology, and although a number of innovative examples have been reported, the emergent properties of systems in which enzymes are coupled together have not been fully explored. Here, we report the self-assembly of a DNA scaffold made of DNA strips that include 'hinges' to which biomolecules can be tethered.

Self-assembly of enzymes on DNA scaffolds: en route to biocatalytic cascades and the synthesis of metallic nanowires.

DNA strands consisting of programmed sequence-specific domains were synthesized by the rolling circle amplification (RCA) process. The spatial positioning of glucose oxidase (GOx) and of horseradish peroxidase (HRP) on the RCA-synthesized DNA template via hybridization enabled the activation of the bienzyme cascade. The GOx-catalyzed oxidation of glucose yielded gluconic acid and H(2)O(2), and the resulting H(2)O(2) oxidized 2,2'-azino-bis[3-ethylbenzthiazoline-6-sulfonic-acid] (ABTS(2-)) in the presence of HRP.

Supramolecular aptamer-thrombin linear and branched nanostructures.

Alpha and beta conjugated bis-aptamers against thrombin act as bidentate "glue" for the self-assembly of thrombin nanowires; mixing the bidentate aptamer with a tripodal tridentate alpha aptamer construct yields branched thrombin nanowire structures.

Probing kinase activities by electrochemistry, contact-angle measurements, and molecular-force interactions.

Three different methods to investigate the activity of a protein kinase (casein kinase, CK2) are described. The phosphorylation of the sequence-specific peptide (1) by CK2 was monitored by electrochemical impedance spectroscopy (EIS). Phosphorylation of the peptide monolayer assembled on a Au electrode yields a negatively charged surface that electrostatically repels the negatively charged redox label [Fe(CN)6]3-/4-, thus increasing the interfacial electron-transfer resistance.

A polycatenated DNA scaffold for the one-step assembly of hierarchical nanostructures.

A unique DNA scaffold was prepared for the one-step self-assembly of hierarchical nanostructures onto which multiple proteins or nanoparticles are positioned on a single template with precise relative spatial orientation. The architecture is a topologically complex ladder-shaped polycatenane in which the "rungs" of the ladder are used to bring together the individual rings of the mechanically interlocked structure, and the "rails" are available for hierarchical assembly, whose effectiveness has been demonstrated with proteins, complementary DNA, and gold nanoparticles. The ability of this template to form from linear monomers and simultaneously bind two proteins was demonstrated by chemical force microscopy, transmission electron microscopy, and confocal fluorescence microscopy.