Scaled and Efficient Derivation of Loss of Function Alleles in Risk Genes for Neurodevelopmental and Psychiatric Disorders in Human iPSC.

Translating genetic findings for neurodevelopmental and psychiatric disorders (NPD) into actionable disease biology would benefit from large-scale and unbiased functional studies of NPD genes. Leveraging the cytosine base editing (CBE) system, here we developed a pipeline for clonal loss-of-function (LoF) allele mutagenesis in human induced pluripotent stem cells (hiPSCs) by introducing premature stop-codons (iSTOP) that lead to mRNA nonsense-mediated-decay (NMD) or protein truncation. We tested the pipeline for 23 NPD genes on 3 hiPSC lines and achieved highly reproducible, efficient iSTOP editing in 22 NPD genes. Using RNAseq, we confirmed their pluripotency, absence of chromosomal abnormalities, and NMD. Interestingly, for three schizophrenia risk genes (SETD1A, TRIO, CUL1), despite the high efficiency of base editing, we only obtained heterozygous LoF alleles, suggesting their essential roles for cell growth. We replicated the reported neural phenotypes of SHANK3-haploinsufficiency and found CUL1-LoF reduced neurite branches and synaptic puncta density. This iSTOP pipeline enables a scaled and efficient LoF mutagenesis of NPD genes, yielding an invaluable shareable resource.

Biomimetic Strategies for Peripheral Nerve Injury Repair: An Exploration of Microarchitecture and Cellularization.

Injuries to the nervous system present formidable challenges to scientists, clinicians, and patients. While regeneration within the central nervous system is minimal, peripheral nerves can regenerate, albeit with limitations. The regenerative mechanisms of the peripheral nervous system thus provide fertile ground for clinical and scientific advancement, and opportunities to learn fundamental lessons regarding nerve behavior in the context of regeneration, particularly the relationship of axons to their support cells and the extracellular matrix environment. However, few current interventions adequately address peripheral nerve injuries. This article aims to elucidate areas in which progress might be made toward developing better interventions, particularly using synthetic nerve grafts. The article first provides a thorough review of peripheral nerve anatomy, physiology, and the regenerative mechanisms that occur in response to injury. This is followed by a discussion of currently available interventions for peripheral nerve injuries. Promising biomaterial fabrication techniques which aim to recapitulate nerve architecture, along with approaches to enhancing these biomaterial scaffolds with growth factors and cellular components, are then described. The final section elucidates specific considerations when developing nerve grafts, including utilizing induced pluripotent stem cells, Schwann cells, nerve growth factors, and multilayered structures that mimic the architectures of the natural nerve.

Deterministic Role of Carbon Nanotube-Substrate Coupling for Ultrahigh Actuation in Bilayer Electrothermal Actuators.

Here, the actuation response of an architectured electrothermal actuator comprising a single layer of carbon nanotube (CNT) film and a relatively thicker film of silk, cellulose, or polydimethylsiloxane is studied. An electric current is passed through the CNT film, which generates heat responsible for electrothermal actuation, in all samples, affixed as per doubly clamped beam configuration. All samples, including pure CNT film, show remarkable actuation such that actuation monotonically increases with the applied voltage. Cyclic pulsed electrical loading shows a lag in the electric current stimulus and the actuation. Remarkably, an ultrahigh actuation of ∼2.8%, which was 72 times more than that shown by pure CNT film, is measured in the CNT-cellulose film, that is, the architectured actuator with the natural polymer having the functional property of hygroexpansion and the structural hierarchy of the CNT film, however, at a significantly larger length scale. Overall, the synergetic contribution of the individual layers in these bilayered actuators enabled achieving ultrahigh electrothermal actuation compared to the homogeneous, synthetic polymer-based devices. A detailed discussion, which also includes examination of the role of the hierarchical substructure and the functional properties of the substrate and numerical analysis using the finite element method, is presented to highlight the actuation mechanism in the fabricated actuators.

Metamaterial-enhanced infrared attenuated total reflection spectroscopy.

The use of Fourier transform infrared spectroscopy with attenuated total reflection (FTIR-ATR) allows solid or liquid samples to be characterised directly without specific sample preparation. In such a system, the evanescent waves generated through total internal reflection within a crystal interact with the sample under test. In this work we explore the use of a mid-infrared metasurface to enhance the interaction between molecular vibrations and the evanescent waves. A complementary ring-resonator structure was patterned onto both silicon and SiO2/Si substrates, and the spectral properties of both devices were characterised using a FTIR-ATR system. Minima in reflectance were observed corresponding to the resonance of the metasurface on the silicon substrate, and to the hybrid resonance of phonon modes and metasurface resonances on the SiO2/Si substrate, in good agreement with simulations. Preliminary experiments were undertaken using mixtures containing trace amounts of butyl acetate diluted with oleic acid. Without the use of a metasurface, the minimum concentration of butyl acetate that could be clearly detected was 10%, whereas the use of the metasurface on the SiO2/Si substrate allowed the detection of 1% butyl acetate. This demonstrates the potential of using metasurfaces to enhance trace chemical detection in FTIR-ATR systems.

Photoresponse of double-stacked graphene to Infrared radiation.

We report the photoresponse of stacked graphene layers towards infrared radiation. Graphene is stacked in two configurations, namely, crossed and parallel layers. Raman analysis demonstrated a strong interaction among the stacked graphene layers. Graphene in the crossed configuration exhibited the presence of both negative and positive conductivities; however, other configurations of graphene exhibited positive conductivity only. The presence of negative photoconductivity is proposed to be due to oxygen or oxygen-related functional group absorbents that are trapped in between two monolayers of graphene and act as scattering centers for free carriers. An interesting trend is reported in differential conductivity when stacked layers are compared with multilayers and parallel-stacked graphene layers.

Enhanced photoresponse in monolayer hydrogenated graphene photodetector.

We report the photoresponse of a hydrogenated graphene (H-graphene)-based infrared (IR) photodetector that is 4 times higher than that of pristine graphene. An enhanced photoresponse in H-graphene is attributed to the longer photoinduced carrier lifetime and hence a higher internal quantum efficiency of the device. Moreover, a variation in the angle of incidence of IR radiation demonstrated a nonlinear photoresponse of the detector, which can be attributed to the photon drag effect. However, a linear dependence of the photoresponse is revealed with different incident powers for a given angle of IR incidence. This study presents H-graphene as a tunable photodetector for advanced photoelectronic devices with higher responsivity. In addition, in situ tunability of the graphene bandgap enables achieving a cost-effective technique for developing photodetectors without involving any external treatments.

Nonlinear optical absorption in a graphene infrared photodetector.

The photoresponse of the graphene photodetector elucidated strong dependence on several optical parameters, such as the angle of incidence and the incident power of infrared exposure at room temperature. The sinusoidal dependence of the photoresponse on incidence angle, which had not been realized before, has now been revealed. The combined effect of the photo excited charge carrier and the photon drag effect explain this nonlinear optical absorption in graphene at lower incident power. The nonlinear dependence of the charge carrier generation on the incident power revealed that this process contributed to the nonlinear photoresponse. However, a deviation is observed at a higher incident power due to the induction of thermal effects in the graphene lattice. This work demonstrates the tunability of the graphene photodetector under a systematic variation that involves both parameters.

Defect-induced enhancement and quenching control of photocurrent in few-layer graphene photodetectors.

A novel approach is presented for achieving an enhanced photoresponse in a few layer graphene (FLG) based photodetector that is realized by introducing defect sites in the FLG. Fabrication induced wrinkle formation in graphene presented a four-fold enhancement in the photocurrent when compared to unfold FLG. Interestingly, it was observed that the addition of few multiwalled carbon nanotubes to an FLG improves the photocurrent by two-fold along with a highly stable response as compared to FLG alone.

Chemical vapor detection using nonlinear electrical properties of carbon nanotube bundles.

We demonstrate the electrical transport behavior of carbon nanotubes (CNTs) upon exposure to organic analytes (namely ethanol, benzene, acetone and toluene). The resulting nonlinear current-voltage characteristics revealed a power law dependence of the differential conductivity on the applied bias voltage. Moreover, suppression of differential conductivity at zero bias is found to be dependent on different selective analytes. The power law exponent values have been monitored before, during and after exposure to the chemicals, which revealed a reversible change in the number of electron conducting channels. Therefore, the reduction in the number of conductive paths can be attributed to the interaction of the chemical analyte on the CNT surfaces, which causes a decrease in the differential conductivity of the CNT sample. These results demonstrate chemical selectivity of CNTs due to varying electronic interaction with different chemical analytes.