Effect of the Phosphoryl Guanidine Modification in Chimeric DNA-RNA crRNAs on the Activity of the CRISPR-Cas9 System In Vitro.

Currently, the CRISPR-Cas9 system serves as a prevalent tool for genome editing and gene expression regulation. Its therapeutic application is limited by off-target effects that can affect genomic integrity through nonspecific, undesirable changes in the genome. Various strategies have been explored to mitigate the off-target effects. Many approaches focus on modifying components of the system, namely, Cas9 and guide RNAs, to enhance specificity. However, a common challenge is that methods aiming to increase specificity often result in a significant reduction in the editing efficiency. Here, we introduce a novel approach to modifying crRNA to balance CRISPR-Cas9 specificity and efficiency. Our approach involves incorporating nucleoside modifications, such as replacing ribo- to deoxyribonucleosides and backbone modifications, using phosphoryl guanidine groups, specifically 1,3-dimethylimidazolidin-2-ylidene phosphoramidate. In this case, within the first 10 nucleotides from the 5' crRNA end, phosphodiester bonds are substituted with phosphoryl guanidine groups. We demonstrate that crRNAs containing a combination of deoxyribonucleosides and single or multiple phosphoryl guanidine groups facilitate the modulation of CRISPR-Cas9 system activity while improving its specificity in vitro.

Influence of Combinations of Lipophilic and Phosphate Backbone Modifications on Cellular Uptake of Modified Oligonucleotides.

Numerous types of oligonucleotide modifications have been developed since automated synthesis of DNA/RNA became a common instrument in the creation of synthetic oligonucleotides. Despite the growing number of types of oligonucleotide modifications under development, only a few of them and, moreover, their combinations have been studied widely enough in terms of their influence on the properties of corresponding NA constructions. In the present study, a number of oligonucleotides with combinations of 3'-end lipophilic (a single cholesteryl or a pair of dodecyl residues) and phosphate backbone modifications were synthesized. The influence of the combination of used lipophilic groups with phosphate modifications of various natures and different positions on the efficiency of cell penetration was evaluated. The obtained results indicate that even a couple of phosphate modifications are able to affect a set of oligonucleotide properties in a complex manner and can remarkably change cellular uptake. These data clearly show that the strategy of using different patterns of modification combinations has great potential for the rational design of oligonucleotide structures with desired predefined properties.

Fork- and Comb-like Lipophilic Structures: Different Chemical Approaches to the Synthesis of Oligonucleotides with Multiple Dodecyl Residues.

Lipophilic oligonucleotide conjugates represent a powerful tool for nucleic acid cellular delivery, and many methods for their synthesis have been developed over the past few decades. In the present study, a number of chemical approaches for the synthesis of different fork- and comb-like dodecyl-containing oligonucleotide structures were performed, including use of non-nucleotide units and different types of phosphate modifications such as alkyl phosphoramidate, phosphoryl guanidine, and triazinyl phosphoramidate. The influence of the number of introduced lipophilic residues, their mutual arrangement, and the type of formed modification backbone on cell penetration was evaluated. The results obtained indicate great potential in the developed chemical approaches, not only for the synthesis of complex oligonucleotide structures but also for the fine-tuning of their properties.

Reconfigurable VO2 metasurfaces with hybrid electro-optical control: manipulating THz radiation with 0.3 W/cm2 light.

Dynamicallyprogrammable metasurfaces capable of manipulating terahertz (THz) wavefronts in various manners depending on external controls are highly desired for next-generation wireless communication systems and new tools for THz diagnostics. Such metasurfaces may utilize the insulator-to-metal transition in V O 2, which can be induced both electrically and optically. Optical control is especially convenient for individual addressing to each meta-atom, but it is hampered by the high optical switching threshold of V O 2. We experimentally realize V O 2-based THz metasurfaces with hybrid electro-optical control when the metasurface is brought close to the transition point by an almost-threshold current, and then is easily switched by unfocused continuous-wave light. We were able to control the metasurface THz transmission by 0.4W/c m 2 near-IR light, while purely optical switching required tightly focused light with an intensity of >3×105 W/c m 2. After correcting for the fact that a tightly focused spot dissipates heat easier, we estimate that the optical switching threshold reduction due to the electric current alone is ∼2 orders of magnitude. Finally, coating the metasurface with Au nanoparticles further reduced the threshold by 30% due to plasmonic effects.

Zero-Bias Photodetection in 2D Materials via Geometric Design of Contacts.

Structural or crystal asymmetry is a necessary condition for the emergence of zero-bias photocurrent in light detectors. Structural asymmetry has been typically achieved via p-n doping, which is a technologically complex process. Here, we propose an alternative approach to achieve zero-bias photocurrent in two-dimensional (2D) material flakes exploiting the geometrical nonequivalence of source and drain contacts. As a prototypical example, we equip a square-shaped flake of PdSe2 with mutually orthogonal metal leads. Upon uniform illumination with linearly polarized light, the device demonstrates nonzero photocurrent which flips its sign upon 90° polarization rotation. The origin of zero-bias photocurrent lies in a polarization-dependent lightning-rod effect. It enhances the electromagnetic field at one contact from the orthogonal pair and selectively activates the internal photoeffect at the respective metal-PdSe2 Schottky junction. The proposed technology of contact engineering is independent of a particular light-detection mechanism and can be extended to arbitrary 2D materials.

The influence of copper ions on the transport and relaxation properties of hydrated eumelanin.

Eumelanin, the human skin pigment, is a poly-indolequinone material possessing a unique combination of physical and chemical properties. For numerous applications, the conductivity of eumelanin is of paramount importance. However, its hydration dependent conductivity is not well studied using transport-relaxation methods. Furthermore, there is no such work taking into account the simultaneous control of humidity as well as metal ion concentration. Here we present the first such study of the transport and relaxation characteristics of synthetic eumelanin doped with various Cu ion concentrations while controlling the humidity with a frequency range of 10-3 Hz-1 MHz. We found that Cu ions do not cause the appearance of additional relaxation processes, but partially slow down those present in neat eumelanin. In addition, considering previously published work, the key relaxation process observed in doped and undoped materials is associated with the moisture-induced synthesis of uncharged semiquinones and a corresponding increase in the overall aromaticity of the material.

Ultralow-noise Terahertz Detection by p-n Junctions in Gapped Bilayer Graphene.

Graphene shows strong promise for the detection of terahertz (THz) radiation due to its high carrier mobility, compatibility with on-chip waveguides and transistors, and small heat capacitance. At the same time, weak reaction of graphene's physical properties on the detected radiation can be traced down to the absence of a band gap. Here, we study the effect of electrically induced band gap on THz detection in graphene bilayer with split-gate p-n junction. We show that gap induction leads to a simultaneous increase in current and voltage responsivities. At operating temperatures of ∼25 K, the responsivity at a 20 meV band gap is from 3 to 20 times larger than that in the gapless state. The maximum voltage responsivity of our devices at 0.13 THz illumination exceeds 50 kV/W, while the noise equivalent power falls down to 36 fW/Hz1/2.

Terahertz Photoconductivity in Bilayer Graphene Transistors: Evidence for Tunneling at Gate-Induced Junctions.

Photoconductivity of novel materials is the key property of interest for design of photodetectors, optical modulators, and switches. Despite the photoconductivity of most novel 2d materials having been studied both theoretically and experimentally, the same is not true for 2d p-n junctions that are necessary blocks of most electronic devices. Here, we study the sub-terahertz photocoductivity of gapped bilayer graphene with electrically induced p-n junctions. We find a strong positive contribution from junctions to resistance, temperature resistance coefficient, and photoresistivity at cryogenic temperatures T ∼ 20 K. The contribution to these quantities from junctions exceeds strongly the bulk values at uniform channel doping even at small band gaps of ∼10 meV. We further show that positive junction photoresistance is a hallmark of interband tunneling, and not of intraband thermionic conduction. Our results point to the possibility of creating various interband tunneling devices based on bilayer graphene, including steep-switching transistors and selective sensors.

Topological phase singularities in atomically thin high-refractive-index materials.

Atomically thin transition metal dichalcogenides (TMDCs) present a promising platform for numerous photonic applications due to excitonic spectral features, possibility to tune their constants by external gating, doping, or light, and mechanical stability. Utilization of such materials for sensing or optical modulation purposes would require a clever optical design, as by itself the 2D materials can offer only a small optical phase delay - consequence of the atomic thickness. To address this issue, we combine films of 2D semiconductors which exhibit excitonic lines with the Fabry-Perot resonators of the standard commercial SiO2/Si substrate, in order to realize topological phase singularities in reflection. Around these singularities, reflection spectra demonstrate rapid phase changes while the structure behaves as a perfect absorber. Furthermore, we demonstrate that such topological phase singularities are ubiquitous for the entire class of atomically thin TMDCs and other high-refractive-index materials, making it a powerful tool for phase engineering in flat optics. As a practical demonstration, we employ PdSe2 topological phase singularities for a refractive index sensor and demonstrate its superior phase sensitivity compared to typical surface plasmon resonance sensors.

Multifunctional Elastic Nanocomposites with Extremely Low Concentrations of Single-Walled Carbon Nanotubes.

Stretchable and flexible electronics has attracted broad attention over the last years. Nanocomposites based on elastomers and carbon nanotubes are a promising material for soft electronic applications. Despite the fact that single-walled carbon nanotube (SWCNT) based nanocomposites often demonstrate superior properties, the vast majority of the studies were devoted to those based on multiwalled carbon nanotubes (MWCNTs) mainly because of their higher availability and easier processing procedures. Moreover, high weight concentrations of MWCNTs are often required for high performance of the nanocomposites in electronic applications. Inspired by the recent drop in the SWCNT price, we have focused on fabrication of elastic nanocomposites with very low concentrations of SWCNTs to reduce the cost of nanocomposites further. In this work, we use a fast method of coagulation (antisolvent) precipitation to fabricate elastic composites based on thermoplastic polyurethane (TPU) and SWCNTs with a homogeneous distribution of SWCNTs in bulk TPU. Applicability of the approach is confirmed by extra low percolation threshold of 0.006 wt % and, as a consequence, by the state-of-the-art performance of fabricated elastic nanocomposites at very low SWCNT concentrations for strain sensing (gauge factor of 82 at 0.05 wt %) and EMI shielding (efficiency of 30 dB mm-1 at 0.01 wt %).

Water-Activated Semiquinone Formation and Carboxylic Acid Dissociation in Melanin Revealed by Infrared Spectroscopy.

Eumelanin is a widespread biomacromolecule pigment in the biosphere and has been widely investigated for numerous bioelectronics and energetic applications. Many of these applications depend on eumelanin's ability to conduct proton current at various levels of hydration. The origin of this behavior is connected to a comproportionation reaction between oxidized and reduced monomer moieties and water. A hydration-dependent FTIR spectroscopic study on eumelanin is presented herein, which allows for the first time tracking the comproportionation reaction via the gradual increase of the overall aromaticity of melanin monomers in the course of hydration. We identified spectral features associated with the presence of specific "one and a half" C𝌁O bonds, typical for o-semiquinones. Signatures of semiquinone monomers with internal hydrogen bonds and that carboxylic groups, in contrast to semiquinones, begin to dissociate at the very beginning of melanin hydration were indicated. As such, we suggest a modification to the common hydration-dependent conductivity mechanism and propose that the conductivity at low hydration is dominated by carboxylic acid protons, whereas higher hydration levels manifest semiquinone protons.

Highly Efficient Piezoelectrets through Ultra-Soft Elastomeric Spacers.

Piezoelectrets are artificial ferroelectrics that are produced from non-polar air-filled porous polymers by symmetry breaking through high-voltage-induced Paschen breakdown in air. A new strategy for three-layer polymer sandwiches is introduced by separating the electrical from the mechanical response. A 3D-printed grid of periodically spaced thermoplastic polyurethane (TPU) spacers and air channels was sandwiched between two thin fluoroethylene propylene (FEP) films. After corona charging, the air-filled sections acted as electroactive elements, while the ultra-soft TPU sections determined the mechanical stiffness. Due to the ultra-soft TPU sections, very high quasi-static (22,000 pC N-1) and dynamic (7500 pC N-1) d33 coefficients were achieved. The isothermal stability of the d33 coefficients showed a strong dependence on poling temperature. Furthermore, the thermally stimulated discharge currents revealed well-known instability of positive charge carriers in FEP, thereby offering the possibility of stabilization by high-temperature poling. The dependences of the dynamic d33 coefficient on seismic mass and acceleration showed high coefficients, even at accelerations approaching that of gravity. An advanced analytical model rationalizes the magnitude of the obtained quasi-static d33 coefficients of the suggested structure indicating a potential for further optimization.

Rotational coherence of encapsulated ortho and para water in fullerene-C60 revealed by time-domain terahertz spectroscopy.

We resolve the real-time coherent rotational motion of isolated water molecules encapsulated in fullerene-C60 cages by time-domain terahertz (THz) spectroscopy. We employ single-cycle THz pulses to excite the low-frequency rotational motion of water and measure the subsequent coherent emission of electromagnetic waves by water molecules. At temperatures below ~ 100 K, C60 lattice vibrational damping is mitigated and the quantum dynamics of confined water are resolved with a markedly long rotational coherence, extended beyond 10 ps. The observed rotational transitions agree well with low-frequency rotational dynamics of single water molecules in the gas phase. However, some additional spectral features with their major contribution at ~2.26 THz are also observed which may indicate interaction between water rotation and the C60 lattice phonons. We also resolve the real-time change of the emission pattern of water after a sudden cooling to 4 K, signifying the conversion of ortho-water to para-water over the course of 10s hours. The observed long coherent rotational dynamics of isolated water molecules confined in C60 makes this system an attractive candidate for future quantum technology.

Aero-Ga2O3 Nanomaterial Electromagnetically Transparent from Microwaves to Terahertz for Internet of Things Applications.

In this paper, fabrication of a new material is reported, the so-called Aero-Ga2O3 or Aerogallox, which represents an ultra-porous and ultra-lightweight three-dimensional architecture made from interconnected microtubes of gallium oxide with nanometer thin walls. The material is fabricated using epitaxial growth of an ultrathin layer of gallium nitride on zinc oxide microtetrapods followed by decomposition of sacrificial ZnO and oxidation of GaN which according to the results of X-ray diffraction (XRD) and X-ray photoemission spectroscopy (XPS) characterizations, is transformed gradually in ß-Ga2O3 with almost stoichiometric composition. The investigations show that the developed ultra-porous Aerogallox exhibits extremely low reflectivity and high transmissivity in an ultrabroadband electromagnetic spectrum ranging from X-band (8-12 GHz) to several terahertz which opens possibilities for quite new applications of gallium oxide, previously not anticipated.

Resonant terahertz detection using graphene plasmons.

Plasmons, collective oscillations of electron systems, can efficiently couple light and electric current, and thus can be used to create sub-wavelength photodetectors, radiation mixers, and on-chip spectrometers. Despite considerable effort, it has proven challenging to implement plasmonic devices operating at terahertz frequencies. The material capable to meet this challenge is graphene as it supports long-lived electrically tunable plasmons. Here we demonstrate plasmon-assisted resonant detection of terahertz radiation by antenna-coupled graphene transistors that act as both plasmonic Fabry-Perot cavities and rectifying elements. By varying the plasmon velocity using gate voltage, we tune our detectors between multiple resonant modes and exploit this functionality to measure plasmon wavelength and lifetime in bilayer graphene as well as to probe collective modes in its moiré minibands. Our devices offer a convenient tool for further plasmonic research that is often exceedingly difficult under non-ambient conditions (e.g. cryogenic temperatures) and promise a viable route for various photonic applications.

Analytical prediction of the piezoelectric d33 response of fluoropolymer arrays with tubular air channels.

The present study is focused on tubular multi-channel arrays composed of commercial fluoropolymer (FEP) tubes with different wall thickness. After proper charging in a high electric field, such tubular structures exhibit a large piezoelectric [Formula: see text] coefficient significantly exceeding the values of classical polymer ferroelectrics and being even comparable to conventional lead-free piezoceramics. The quasistatic piezoelectric [Formula: see text] coefficient was theoretically derived and its upper limits were evaluated considering charging and mechanical properties of the arrays. In order to optimize the [Formula: see text] coefficient the remanent polarization and the mechanical properties were taken into account, both being strongly dependent on the air channel geometry as well as on the wall thickness of the FEP tubes. The model predictions are compared with experimental d33 coefficients for two particular arrays with equal air gaps of 250 µm, but with different wall thickness of utilized FEP tubes of 50 µm and 120 µm, respectively. Analytical modeling allows for the prediction that arrays made of FEP tubes with a wall thickness of 10 µm are foreseen to exhibit a superb piezoelectric response of up to 600 pC/N if the height of stadium-like shaped air channels is reduced down to 50 µm, making them potentially interesting for application as highly sensitive sensors and energy harvesting.