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Ensuring sufficient gRNA transcript levels is critical for obtaining optimal CRISPR-Cas9 gene editing efficiency. The standard gRNA scaffold contains a sequence of four thymine nucleotides (4T), which is known to inhibit transcription from Pol III promoters such as the U6 promoter. Our study showed that using standard plasmid transfection protocols, the presence of these 4Ts did not significantly affect editing efficiency, as most of the gRNAs tested (55 gRNAs) achieved near-perfect editing outcomes. We observed that gRNAs with lower activity were T-rich and had reduced gRNA transcript levels. However, this issue can be effectively resolved by increasing transcript levels, which can be readily achieved by shortening the 4T sequences. In this study, we demonstrated this by modifying the sequences to 3TC. Although the 3TC scaffold modification did not improve editing efficiency for already efficient gRNAs when high vector quantities were available, it proved highly beneficial under conditions of limited vector availability, where the 3TC scaffold yielded higher editing efficiency. Additionally, we demonstrated that the 3TC scaffold is compatible with SpCas9 high-fidelity variants and ABEmax base editing, enhancing their editing efficiency. Another commonly used natural Cas9 variant, SaCas9, also benefited from the 3TC scaffold sequence modification, which increased gRNA transcription and subsequently improved editing activity. This modification was applied to the EDIT-101 therapeutic strategy, where it demonstrated marked improvements in performance. This study highlights the importance of shortening the 4T sequences in the gRNA scaffold to optimize gRNA transcript expression for enhanced CRISPR-Cas9 gene editing efficiency. This optimization is particularly important for therapeutic applications, where the quantity of vector is often limited, ensuring more effective and optimal outcomes.

A neural network, BES-ELMnet, predicting a quasi-periodic disruptive eruption of the plasma energy and particles known as edge localized mode (ELM) onset is developed with observed pedestal turbulence from the beam emission spectroscopy system in DIII-D. BES-ELMnet has convolutional and fully-connected layers, taking two-dimensional plasma fluctuations with a temporal window of size 128 µs and generating a scalar output which can be interpreted as a probability of the upcoming ELM onset. As approximately labeled inter-ELM broadband ( 15kHz⩽f⩽150kHz ) fluctuations are given to the network, BES-ELMnet learns by itself ELM-related precursors arising before the onsets through supervised learning scheme. BES-ELMnet achieves the gradually increasing ELM onset probabilities between two consecutive ELMs during the inter-ELM phases and can forecast the first ELM onsets which occur after the high confinement mode transition. We further investigate the network generality in terms of the selected frequency band to ensure the use of BES-ELMnet for various operation regimes without changing the trained architecture. Therefore, our novel prediction method will enhance a proactive high confinement mode control of fusion-grade plasmas.

The optimization of helically symmetric experiment (HSX) for reduced microinstability has been achieved by examining a large set of configurations within a neighborhood of the standard operating configuration. This entailed generating a database of more than 10 6 magnetic-field configurations for HSX by varying the currents in external coils. Using a set of volume-averaged metrics and gyrokinetic simulations, this database has helped to identify a set of configurations that can be used to regulate trapped-electron-mode stability in HSX. This set of configurations is also found to correlate flux-surface elongation and triangularity with an increase in magnetic-well depth, an increase in rotational transform, and low neoclassical heat-flux relative to the standard quasi-helically-symmetric configuration. These results demonstrate sensitivity of plasma behavior in response to changes in a 3D magnetic field to both neoclassical and gyrokinetic models, and the experimental potential in HSX to explore turbulence optimization. This perturbative optimization approach is not unique to HSX, and can readily be deployed on existing fusion devices to identify novel magnetic-fields to be used in turbulence-optimization experiments.

This paper presents the first results from the analysis of Balmer-alpha spectra at Wendelstein 7-X which contain the broad charge exchange emission from fast-ions. The measured spectra are compared to synthetic spectra predicted by the FIDASIM code, which has been supplied with the 3D magnetic fields from VMEC, 5D fast-ion distribution functions from ASCOT, and a realistic Neutral Beam Injection geometry including beam particle blocking elements. Detailed modeling of the beam emission shows excellent agreement between measured beam emission spectra and predictions. In contrast, modeling of beam halo radiation and Fast-Ion H-Alpha signals (FIDA) is more challenging due to strong passive contributions. While about 50% of the halo radiation can be attributed to passive signals from edge neutrals, the FIDA emission—in particular for an edge-localized line of sights—is dominated by passive emission. This is in part explained by high neutral densities in the plasma edge and in part by edge-born fast-ion populations as demonstrated by detailed modeling of the edge fast-ion distribution.

The Wisconsin high-temperature superconductor axisymmetric mirror experiment (WHAM) will be a high-field platform for prototyping technologies, validating interchange stabilization techniques and benchmarking numerical code performance, enabling the next step up to reactor parameters. A detailed overview of the experimental apparatus and its various subsystems is presented. WHAM will use electron cyclotron heating to ionize and build a dense target plasma for neutral beam injection of fast ions, stabilized by edge-biased sheared flow. At 25 keV injection energies, charge exchange dominates over impact ionization and limits the effectiveness of neutral beam injection fuelling. This paper outlines an iterative technique for self-consistently predicting the neutral beam driven anisotropic ion distribution and its role in the finite beta equilibrium. Beginning with recent work by Egedal et al. (Nucl. Fusion, vol. 62, no. 12, 2022, p. 126053) on the WHAM geometry, we detail how the FIDASIM code is used to model the charge exchange sources and sinks in the distribution function, and both are combined with an anisotropic magnetohydrodynamic equilibrium solver method to self-consistently reach an equilibrium. We compare this with recent results using the CQL3D code adapted for the mirror geometry, which includes the high-harmonic fast wave heating of fast ions.

Microtearing modes (ω ~ ω de , k θ ρ s ~ 0.1, k r ρ s ~ 0.3-1.0) are predicted to cause substantial electron thermal transport in the H-mode pedestal, limiting fusion performance on high-power devices. Localized 2D measurements of low-to-intermediate wavenumber density fluctuations in the edge of DIII-D H-mode plasmas suggest the presence of temporally intermittent or \"bursting\" microtearing modes during the inter-ELM cycle in the steep gradient region. These measurements are obtained using a new intermediate radial resolution multichannel Charge eXchange Imaging (CXI) diagnostic that analyzes the intensity of the CVI emission at 529 nm. The mode is peaked near ρ=0.97 with a radial full-width half maximum of 1.5-2.0 cm and a normalized density fluctuation amplitude peak of dn C /n C ~5%, peaking near 60 kHz in the lab frame. The mode additionally features a broad spectral width from 30-180 kHz and propagates in the electron diamagnetic direction in the plasma frame. CXI measurements taken during discharges with high density and current ramps provide evidence that microtearing modes become unstable at finite collisionality and remain at nearly the same location in ρ-space despite scanning of rational surface position. This indicates that the location of the peak in the electron diamagnetic frequency is a stronger driving force than individual rational surfaces.

A real-time detection of the plasma confinement regime can enable new advanced plasma control capabilities for both the access to and sustainment of enhanced confinement regimes in fusion devices. For example, a real-time indication of the confinement regime can facilitate transition to the high-performing wide-pedestal (WP) quiescent H-mode, or avoid unwanted transitions to lower confinement regimes that may induce plasma termination. To demonstrate real-time confinement regime detection, we use the 2D beam emission spectroscopy (BES) diagnostic system to capture localized density fluctuations of long wavelength turbulent modes in the edge region at a 1 MHz sampling rate. BES data from 330 discharges in either L-mode, H-mode, quiescent H (QH)-mode, or WP QH-mode were collected from the DIII-D tokamak and curated to develop a high-quality database to train a deep-learning classification model for real-time confinement detection. We utilize the 6×8 spatial configuration with a time window of 1024 µ s and recast the input to obtain spectral-like features via fast Fourier transform preprocessing. We employ a shallow 3D convolutional neural network for the multivariate time-series classification task and utilize a softmax in the final dense layer to retrieve a probability distribution over the different confinement regimes. Our model classifies the global confinement state on 44 unseen test discharges with an average F 1 score of 0.94, using only ∼1 ms snippets of BES data at a time. This activity demonstrates the feasibility for real-time data analysis of fluctuation diagnostics in future devices such as ITER, where the need for reliable and advanced plasma control is urgent.

Launched on 2 June 2003 and arriving at Mars on 25 December 2003 after a 7-month interplanetary cruise, Mars Express was the European Space Agency’s first mission to arrive at another planet. After more than 20 years in orbit, the spacecraft and science payload remain in good health and the mission has become the second oldest operational planetary orbiter after Mars Odyssey. This contribution summarizes the Mars Express mission operations, science planning and data archiving systems, processes, and teams that are necessary to run the mission, plan the scientific observations, and execute all necessary commands. It also describes the data download, the ground processing and distribution to the scientific community for the study and analysis of Mars sub-surface, surface, atmosphere, magnetosphere, and moons. This manuscript also describes the main challenges throughout the history of the mission, including several potentially mission-ending anomalies. We summarize the evolution of the ground segment to provide new capabilities not envisaged before launch, whilst simultaneously maintaining or even increasing the quality and quantity of scientific data generated.

Forces exerted by stationary cells have been investigated on the level of single focal adhesions by combining elastic substrates, fluorescence labeling of focal adhesions, and the assumption of localized force when solving the inverse problem of linear elasticity theory. Data simulation confirms that the inverse problem is ill-posed in the presence of noise and shows that in general a regularization scheme is needed to arrive at a reliable force estimate. Spatial and force resolution are restricted by the smoothing action of the elastic kernel, depend on the details of the force and displacement patterns, and are estimated by data simulation. Corrections arising from the spatial distribution of force and from finite substrate size are treated in the framework of a force multipolar expansion. Our method is computationally cheap and could be used to study mechanical activity of cells in real time.

The ExoMars Trace Gas Orbiter (TGO) Science Ground Segment (SGS), comprised of payload Instrument Team, ESA and Russian operational centres, is responsible for planning the science operations of the TGO mission and for the generation and archiving of the scientific data products to levels meeting the scientific aims and criteria specified by the ESA Project Scientist as advised by the Science Working Team (SWT). The ExoMars SGS builds extensively upon tools and experience acquired through earlier ESA planetary missions like Mars and Venus Express, and Rosetta, but also is breaking ground in various respects toward the science operations of future missions like BepiColombo or JUICE. A productive interaction with the Russian partners in the mission facilitates broad and effective collaboration. This paper describes the global organisation and operation of the SGS, with reference to its principal systems, interfaces and operational processes.