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At least 163 records · Page 9

Measuring Thickness-Dependent Relative Light Yield and Detection Efficiency of Scintillator Screens

Digital camera-based neutron imaging systems consisting of a neutron scintillator screen optically coupled to a digital camera are the most common digital neutron imaging system used in the neutron imaging community and are available at any state-of-the-art imaging facility world-wide. Neutron scintillator screens are the integral component of these imaging system that directly interacts with the neutron beam and dictates the neutron capture efficiency and image quality limitations of the imaging system. This work describes a novel approach for testing neutron scintillators that provides a simple and efficient way to measure relative light yield and detection efficiency over a range of scintillator thicknesses using a single scintillator screen and only a few radiographs. Additionally, two methods for correlating the screen thickness to the measured data were implemented and compared. An example 6LiF:ZnS scintillator screen with nominal thicknesses ranging from 0–300 μm was used to demonstrate this approach. The multi-thickness screen and image and data processing methods are not exclusive to neutron scintillator screens but could be applied to X-ray imaging as well. This approach has the potential to benefit the entire radiographic imaging community by offering an efficient path forward for manufacturers to develop higher-performance scintillators and for imaging facilities and service providers to determine the optimal screen parameters for their particular beam and imaging system.

42 ENGINEERING↗

Polarization consistent dielectric screening in polarizable continuum model calculations of solvation energies

A polarization consistent framework, where dielectric screening is affected consistently in polarizable continuum model (PCM) calculations, is employed for the study of solvation energies. The computational framework combines a screened range-separated-hybrid functional (SRSH) with PCM calculations, SRSH-PCM, where dielectric screening is imposed in both PCM self-consistent reaction field (SCRF) iterations and the electronic structure Hamiltonian. We begin by demonstrating the impact of modifying the Hamiltonian to include such dielectric screening in SCRF iterations by considering the solutions of electrostatically embedded Hartree–Fock (HF) exact exchange equations. Long-range screened HF-PCM calculations are shown to capture properly the linear dependence of gap energy of frontier orbitals on the inverse of the dielectric constant, whereas unscreened HF-PCM orbital energies are fallaciously semi-constant with respect to the dielectric constant and, therefore, inconsistent with the ionization energy gaps. Similar trends affect density functional theory (DFT) calculations that aim to achieve predictive quality. Importantly, the dielectric screened calculations are shown to significantly affect DFT- and HF PCM-based solvation energies, where screened solvation energies are smaller compared to the unscreened values. Importantly, SRSH-PCM, therefore, appears to reduce the tendency of DFT-PCM to overestimate solvation energies, where we find the effect to increase with the dielectric constant and the polarity of the molecular solute, trends that enhance the quality of DFT-PCM calculations of solvation energy. Understanding the relationship of dielectric screening in the Hamiltonian and DFT-PCM calculations can ultimately benefit on-going efforts for the design of predictive and parameter free descriptions of solvation energies.

Chemistry↗

Nonlinear screening of dust grains and structurization of dusty plasma

A review of theoretical ideas on the physics of structurization instability of a homogeneous dusty plasma, i.e., the formation of zones with elevated and depressed density of dust grains and their arrangement into different structures observed in laboratory plasma under microgravity conditions, is presented. Theoretical models of compact dust structures that can form in the nonlinear stage of structurization instability, as well as models of a system of voids (both surrounding a compact structure and formed in the center of the structure), are discussed. Two types of structures with very different dimensions are possible, namely, those smaller or larger than the characteristic mean free path of ions in the plasma flow. Both of them are characterized by relatively regular distributions of dust grains; however, the first ones usually require external confinement, while the structures of the second type can be self-sustained (which is of particular interest). In this review, they are called dust clusters and self-organized dust structures, respectively. Both types of the structures are characterized by new physical processes that take place only in the presence of the dust component. The role of nonlinearities in the screening of highly charged dust grains that are often observed in modern laboratory experiments turns out to be great, but these nonlinearities have not received adequate study as of yet. Although structurization takes place upon both linear and nonlinear screening, it can be substantially different under laboratory and astrophysical conditions. Studies on the nonlinear screening of large charges in plasma began several decades ago; however, up to now, this effect was usually disregarded when interpreting the processes occurring in laboratory dusty plasma. One of the aims of the present review was to demonstrate the possibility of describing the nonlinear screening of individual grains and take it into account with the help of the basic equations for the equilibrium between plasma components when analyzing equilibrium structures. The effect of plasma screening nonlinearity on both the diffusion processes and the forces of dust drag by plasma fluxes is analyzed. It is shown how self-organized dust structures form in these processes. In the limit of very small dust grain charges, the forces acting on the dusty plasma components and the set of basic equations for stationary dust structures (with allowance for nonlinear screening) take a standard form. New qualitative effects, such as the suppression of diffusion due to ion scattering from dust grains and the formation of structures of different configurations, are described. A detailed comparison with previous results is performed. It is shown that the solution of basic nonlinear equations for dust structures yields new qualitative effects. A number of new effects to be studied in future dusty plasma experiments with the formation of structures in spherical chambers are predicted (it is assumed that diffusion will play a significant role under microgravity conditions). Recent ground-based experiments, as well as experiments carried out onboard the International Space Station, directly confirm the nonlinear character of screening and the significant role played by this nonlinearity in the structurization of dusty plasma. Experiments on the formation of structures consisting of smaller dust grains within structures formed of larger grains are discussed. It is shown that those experiments can be interpreted only using the concept of nonlinear screening.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Effect of transient liquid flow on retention characteristics of screen acquisition systems

A design analysis, is developed based on experimental data, to predict the effects of transient flow and pressure surges (caused either by valve or pump operation, or by boiling of liquids in warm lines) on the retention performance of screen acquisition systems. A survey of screen liquid acquisition system applications was performed to determine appropriate system environment and classification. A screen model was developed which assumed that the screen device was a uniformly distributed composite orthotropic structure, and which accounted for liquid inflow/outflow, gas ingestion quality, screen stress, and liquid spill. A series of 177 tests using 13 specimens (5 screen meshes, 4 screen device construction/backup methods, and 2 orientations) with three test fluids (isopropyl alcohol, Freon 114, and LH2) provided data which verified important features of the screen model and resulted in a design tool which could accurately predict the transient startup performance acquisition devices.

Cady, E. C.↗

Effect of Vibration on Retention Characteristics of Screen Acquisition Systems

An analytical and experimental investigation of the effect of vibration on the retention characteristics of screen acquisition systems was performed. The functioning of surface tension devices using fine-mesh screens requires that the pressure differential acting on the screen be less than its pressure retention capability. When exceeded, screen breakdown will occur and gas-free expulsion of propellant will no longer be possible. An analytical approach to predicting the effect of vibration was developed. This approach considers the transmission of the vibration to the screens of the device and the coupling of the liquid and the screen in establishing the screen response. A method of evaluating the transient response of the gas/liquid interface within the screen was also developed.

Tegart, J. R.↗

The screen hole plasma sheath of an ion accelerator system

Results of the first probing of the screen hole sheath of an ion accelerator system are presented. The screen hole sheath, represented as a set of equipotential contours, extends over a large distance within the discharge plasma. Under no conditions examined does the sheath enter the screen hole. Edge hole defocusing of multiaperture accelerator systems is due primarily to local plasma density variations rather than adjacent screen hole sheath interactions. The sheath boundary is independent of screen-to-accelerator grid spacing when the grid set is operated at the minimum ion beam divergence condition. Significant ion focusing effects occur in the sheath adjacent to the screen grid webbing leading to increased ion source beam current efficiency with decreasing screen-to-accelerator grid separation and/or screen grid thickness.

Aston, G.↗

Measurement of Screen-Size Effects on Intensity, Scale, and Spectrum of Turbulence in a Free Subsonic Jet

The results are reported of hot-wire anemometer measurements of the fluctuating longitudinal component of the turbulent velocities in the mean flow downstream of screens in an air jet. These measurements have been analyzed by well-established techniques to give the influence of tile screen mesh size on the turbulent intensity, scale, and the power-spectral-density. The results show a linear dependence of the intensity upon the screen mesh size for locations within the central core of the air jet. The spectral-density curves show that the screens redistribute the turbulent energy from the low frequencies (<1000 cps) to the high frequencies (>1000 cps). The effects of the screens are overwhelmed in the mixing region of the jet flow by the turbulence levels existing there. The large pressure drops occurring across the screens reduce the velocity of the jet as compared to the jet without screens by approximately one-third for the velocity and range of mesh sizes investigated and reported in this report. The turbulence scale is a linear function of distance from the nozzle exit and is somewhat greater than comparable jets without screens.

Howard, Charles D.↗

An Automated Algorithm to Screen Massive Training Samples for a Global Impervious Surface Classification

An algorithm is developed to automatically screen the outliers from massive training samples for Global Land Survey - Imperviousness Mapping Project (GLS-IMP). GLS-IMP is to produce a global 30 m spatial resolution impervious cover data set for years 2000 and 2010 based on the Landsat Global Land Survey (GLS) data set. This unprecedented high resolution impervious cover data set is not only significant to the urbanization studies but also desired by the global carbon, hydrology, and energy balance researches. A supervised classification method, regression tree, is applied in this project. A set of accurate training samples is the key to the supervised classifications. Here we developed the global scale training samples from 1 m or so resolution fine resolution satellite data (Quickbird and Worldview2), and then aggregate the fine resolution impervious cover map to 30 m resolution. In order to improve the classification accuracy, the training samples should be screened before used to train the regression tree. It is impossible to manually screen 30 m resolution training samples collected globally. For example, in Europe only, there are 174 training sites. The size of the sites ranges from 4.5 km by 4.5 km to 8.1 km by 3.6 km. The amount training samples are over six millions. Therefore, we develop this automated statistic based algorithm to screen the training samples in two levels: site and scene level. At the site level, all the training samples are divided to 10 groups according to the percentage of the impervious surface within a sample pixel. The samples following in each 10% forms one group. For each group, both univariate and multivariate outliers are detected and removed. Then the screen process escalates to the scene level. A similar screen process but with a looser threshold is applied on the scene level considering the possible variance due to the site difference. We do not perform the screen process across the scenes because the scenes might vary due to the phenology, solar-view geometry, and atmospheric condition etc. factors but not actual landcover difference. Finally, we will compare the classification results from screened and unscreened training samples to assess the improvement achieved by cleaning up the training samples. Keywords:

Tan, Bin↗

Impact of the Screen Configurations on the Estimated Terra MODIS SD Degradation

The Moderate Resolution Imaging Spectroradiometer (MODIS) instruments on-board the Aqua and Terra spacecraft have collected valuable Earth data for the last 19 and 21 years, respectively. MODIS is equipped with various on-orbit calibrators, including a solar diffuser (SD) and solar diffuser stability monitor (SDSM), that are used to monitor changes in the instrument’s gain over time. Nominally, SD calibrations alternate between two configurations: screen open and screen closed. Terra MODIS, however, experienced an anomaly in 2003, which has left the SD door and screen in a permanent open-closed configuration. This resulted in accelerated degradation of the SD on Terra MODIS due to the direct solar radiation exposure every orbit. It also led to an unexpected divergence between the calibration coefficients, m1, or the inverse of the gain, generated using SD data and those generated using lunar observations at the time of the 2003 anomaly. This paper examines the effect of the screen configuration on the Terra SD degradation by analyzing Terra SDSM data and comparing to Aqua. We generate “pseudo-open” Terra SDSM data by modeling the ratio of Aqua screen open and screen closed SDSM data and calculate the Terra SD degradation using this new screen open data. We then examine the effects on our m1 values calculated using this new pseudo-open SD degradation for Terra. Implementing this new degradation results in a smaller discrepancy between the lunar and SD m1 values after the door anomaly, indicating that there may be a systematic error in the SD degradation calculated with screen closed data.

MODIS↗

acCRISPR: an activity-correction method for improving the accuracy of CRISPR screens

Abstract High throughput CRISPR screens are revolutionizing the way scientists unravel the genetic underpinnings of engineered and evolved phenotypes. One of the critical challenges in accurately assessing screening outcomes is accounting for the variability in sgRNA cutting efficiency. Poorly active guides targeting genes essential to screening conditions obscure the growth defects that are expected from disrupting them. Here, we develop acCRISPR, an end-to-end pipeline that identifies essential genes in pooled CRISPR screens using sgRNA read counts obtained from next-generation sequencing. acCRISPR uses experimentally determined cutting efficiencies for each guide in the library to provide an activity correction to the screening outcomes via calculation of an optimization metric, thus determining the fitness effect of disrupted genes. CRISPR-Cas9 and -Cas12a screens were carried out in the non-conventional oleaginous yeast Yarrowia lipolytica and acCRISPR was used to determine a high-confidence set of essential genes for growth under glucose, a common carbon source used for the industrial production of oleochemicals. acCRISPR was also used in screens quantifying relative cellular fitness under high salt conditions to identify genes that were related to salt tolerance. Collectively, this work presents an experimental-computational framework for CRISPR-based functional genomics studies that may be expanded to other non-conventional organisms of interest.

59 BASIC BIOLOGICAL SCIENCES↗

Assessment of Screen-Covered Grooved Sodium Heat Pipes for Microreactor Applications

New nuclear reactor designs that incorporate heat pipes are being investigated for possible near-term deployment in terrestrial applications. Here, this study explores the use of screen-covered axially grooved sodium heat pipes and their applicability for providing heat removal for microreactors. A sodium working fluid is appropriate for microreactors operating in the 5 to 20 MW(thermal) range at approximately 650°C. HTPIPE, a legacy software code, was validated for the case of screen-covered grooves and used to perform steady-state analyses to determine the performance limits of a proposed heat pipe design. The performance limits of a sodium heat pipe with a screen-covered square grooved wick structure is compared to that of an equivalent heat pipe with an annular wick. In a horizontal orientation at an operating temperature of 650°C,the performance limits for the heat pipe with an annular wick configuration are 15% higher than for the screen-covered grooved wick. At operating temperatures below 777°C, the annular wick outperforms the screen-covered grooved wick, and at temperatures above 777°C, the screen-covered grooved wick outperforms the annular wick. However, the marginal performance gain at higher temperatures may not justify the use of heat pipes with a screen-covered grooved wick structure due to increased manufacturing costs.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗

Impurity screening behavior of the high-field side scrape-off layer in near-double-null configurations: prospect for mitigating plasma–material interactions on RF actuators and first-wall components

The impurity screening response of the high-field side (HFS) scrape-off layer (SOL) to localized nitrogen injection is investigated on Alcator C-Mod for magnetic equilibria spanning lower-single-null, double-null and upper-single-null configurations under otherwise identical plasma conditions. L-mode, EDA H-mode and I-mode discharges are investigated. HFS impurity screening is found to depend on magnetic flux balance and the direction of B × $\nabla$B relative to the most active divertor. Impurity 'plume' emission patterns indicate that both parallel and perpendicular (E × B) flows in the SOL contribute to the 'flushing' of impurities towards the active divertor, thereby affecting the overall impurity screening behavior. Despite the fact that the HFS SOL is extremely narrow in near-double-null configurations, this SOL is able to screen locally injected nitrogen at least as effectively as the low-field side (LFS) SOL—up to a factor of 10 more effective, depending on specific plasma conditions and whether the magnetic geometry produces parallel flows that work with or against E × B flows. For situations in which the E × B drift of the impurity ions opposes parallel flow toward the primary divertor, HFS impurity screening is found to be least effective. When E × B drifts assist parallel flow toward the primary divertor, HFS impurity screening is found to be very effective. These data support the idea of placing RF actuators and close-fitting wall components on the HFS of the tokamak. Finally, with this configuration, near-double-null magnetic topologies may be used for active control of plasma parameters at the antenna/plasma interface for optimal RF coupling, mitigate the generation of local impurities from plasma–material interactions and, taking advantage of favorable plasma flows and good screening properties of the HFS SOL, further minimize the impact of wall-born impurity sources on the plasma core.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Investigation of the Effect of Gate Oxide Screening with Adjustment Pulse on Commercial SiC Power MOSFETs

This paper presents a method to recover the negative threshold voltage shift during high field gate oxide screening of 1.2 kV 4H-SiC MOSFETs with an additional adjustment gate voltage pulse. To reduce field failure rates of the MOSFETs in operation, manufacturers perform a screening treatment to remove devices with extrinsic defects in the oxide. Current gate oxide screening procedures are limited to oxide fields at or below ~9 MV/cm for short durations (<1 s), which is not enough to remove all the devices with extrinsic defects. The results show that by implementing a lower field gate pulse, the threshold voltage shift can be partially recovered, and therefore the maximum screening field and time can be increased. However, both the initial screening pulse and the adjustment pulse require careful calibration to prevent significant degradation of the device threshold voltage, on-resistance, interface state density, or intrinsic lifetime. With a well calibrated set of pulses, higher screening fields can be utilized without significantly damaging the devices. This leads to an improvement in the overall screening efficiency of the process, reducing the number of devices with extrinsic oxide defects entering the field, and improving the reliability of the SiC MOSFETs in operation.

42 ENGINEERING↗

A Screening Method Using Pulsed-Power Combined with Infrared Imaging to Detect Pattern Defects in Bulk Metal Foil or Thin Film Resistors

Bulk metal foil and thin film resistors occasionally contain localized defects within the etched resistor pattern. Common defects of this type include in-line constrictions referred to as notches, unremoved resistor material bridges between adjacent pattern lines, and embedded non-conductive particles in the resistor material. Such defects are prone to fracture due to thermomechanical fatigue during powered operation, especially power cycling, resulting in positive resistance change and open circuit failure modes. Common screening methods of optical microscopy, short time overload power tests (e.g., 6.25x rated power for 5 seconds) and burn-in (e.g., 1.5x rated power for 100 hours) are useful, but they are not always effective at removing devices with such defects. An improved method has been developed to screen for localized resistor pattern defects. The method involves the application of brief, high power electrical pulses at a low duty cycle while inspecting the resistor with a high resolution, high speed infrared camera. The following test conditions were found to be suitable for this purpose: 6.25x rated power, 1 to 5 pulses, 50 ms pulse width and 10% duty cycle. During the power pulsing, localized constrictions in the resistor pattern are identifiable as hot spots via the infrared camera. Reject criteria can be established based on observations of hot spots. To assess its effectiveness, a total of two hundred eighty (280) surface mount foil resistors (40 each from 7 different lots) were screened using this infrared technique. The screening identified twenty-nine (29) resistors with significant hot spots in the resistor pattern. All 280 resistors were then subjected to an industry standard 10,000 hour life test at 1x rated power at 70°C with power cycled for 90 minutes on and 30 minutes off. During life testing, three resistors exhibited positive resistance change failure mode. All three failures were due to thermomechanical fatigue failure of localized bridge defects at the locations identified via infrared screening. The results of this evaluation illustrate the benefits of the pulsed-power infrared detection screening technique to identify reliability suspect foil or thin film resistors that may escape common screening methods.

screening↗

A Screening Method Using Pulsed-Power Combined with Infrared Imaging to Detect Pattern Defects in Bulk Metal Foil or Thin Film Resistors

Bulk metal foil and thin film resistors occasionally contain localized defects within the etched resistor pattern. Common defects of this type include in-line constrictions referred to as notches, unremoved resistor material bridges between adjacent pattern lines, and embedded non-conductive particles in the resistor material. Such defects are prone to fracture due to thermomechanical fatigue during powered operation, especially power cycling, resulting in positive resistance change and open circuit failure modes. Common screening methods of optical microscopy, short time overload power tests (e.g., 6.25x rated power for 5 seconds) and burn-in (e.g., 1.5x rated power for 100 hours) are useful, but they are not always effective at removing devices with such defects. An improved method has been developed to screen for localized resistor pattern defects. The method involves the application of brief, high power electrical pulses at a low duty cycle while inspecting the resistor with a high resolution, high speed infrared camera. The following test conditions were found to be suitable for this purpose: 6.25x rated power, 1 to 5 pulses, 50 ms pulse width and 10% duty cycle. During the power pulsing, localized constrictions in the resistor pattern are identifiable as hot spots via the infrared camera. Reject criteria can be established based on observations of hot spots. To assess its effectiveness, a total of two hundred eighty (280) surface mount foil resistors (40 each from 7 different lots) were screened using this infrared technique. The screening identified twenty-nine (29) resistors with significant hot spots in the resistor pattern. All 280 resistors were then subjected to an industry standard 10,000 hour life test at 1x rated power at 70°C with power cycled for 90 minutes on and 30 minutes off. During life testing, three resistors exhibited positive resistance change failure mode. All three failures were due to thermomechanical fatigue failure of localized bridge defects at the locations identified via infrared screening. The results of this evaluation illustrate the benefits of the pulsed-power infrared detection screening technique to identify reliability suspect foil or thin film resistors that may escape common screening methods.

screening↗

Adverse physiological and psychological effects of screen time on children and adolescents: Literature review and case study

A growing body of literature is associating excessive and addictive use of digital media with physical, psychological, social and neurological adverse consequences. Research is focusing more on mobile devices use, and studies suggest that duration, content, after-dark-use, media type and the number of devices are key components determining screen time effects. Physical health effects: excessive screen time is associated with poor sleep and risk factors for cardiovascular diseases such as high blood pressure, obesity, low HDL cholesterol, poor stress regulation (high sympathetic arousal and cortisol dysregulation), and Insulin Resistance. Other physical health consequences include impaired vision and reduced bone density. Psychological effects: internalizing and externalizing behavior is related to poor sleep. Depressive symptoms and suicidal are associated to screen time induced poor sleep, digital device night use, and mobile phone dependency. ADHD-related behavior was linked to sleep problems, overall screen time, and violent and fast-paced content which activates dopamine and the reward pathways. Early and prolonged exposure to violent content is also linked to risk for antisocial behavior and decreased prosocial behavior. Psychoneurological effects: addictive screen time use decreases social coping and involves craving behavior which resembles substance dependence behavior. Brain structural changes related to cognitive control and emotional regulation are associated with digital media addictive behavior. A case study of a treatment of an ADHD diagnosed 9-year-old boy suggests screen time induced ADHD-related behavior could be inaccurately diagnosed as ADHD. Screen time reduction is effective in decreasing ADHD-related behavior.

60 APPLIED LIFE SCIENCES↗

A review of neutronics and thermal hydraulics–based screening methods applied to accelerated nuclear fuel qualification

This paper reviews the state-of-the-art engineering approach for using thermal hydraulic (TH) and neutronics modeling and simulation (M&S) tools to perform rapid screening studies of novel nuclear fuel concepts within the context of accelerated fuel qualification. Global research efforts have introduced nuclear fuel and material concepts that mark a significant departure from traditional reactor materials. The number of new technologies being considered for development for light water reactors and advanced reactor types has created the need for an accelerated fuel qualification procedure. A key component of this procedure is the rapid identification of the most promising fuel concepts using computational screening studies. Advanced TH and neutronic M&S tools should be leveraged to efficiently determine whether the reactor performance and safety characteristics of a given concept warrant additional studies or whether the concept requires modification or elimination. This paper reviews best practices for performing these TH and neutronics screening studies at various stages during a fuel concept's progression through the qualification procedure. The motivation behind standardizing this approach is to minimize time and resources spent on qualification activities for fuel concepts that could be quickly refined or eliminated from consideration based on their reactor physics and TH characteristics. Adoption of this screening procedure—which focuses primarily on nuclear fuels but may be applicable to other reactor materials—will also help accelerate new material qualification by generating boundary conditions crucial to fuel performance evaluations and highlighting needed areas of separate effects experimentation. This article reviews the motivation behind the introduction of novel nuclear fuel concepts, provides incentive for utilizing TH- and neutronics-based screening studies, describes the screening approach and methodology, and includes discussion on how to interpret screening results to provide recommendations for the continued development of a given concept.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Coupling High-Throughput and Targeted Screening for Identification of Nonobvious Metabolic Engineering Targets

Identification of metabolic engineering targets is a fundamental challenge in strain development programs. While high-throughput (HTP) genetic engineering methodologies capable of generating vast diversity are being developed at a rapid rate, a majority of industrially interesting molecules cannot be screened at sufficient throughput to leverage these techniques. We propose a workflow that couples HTP screening of common precursors (e.g., amino acids) that can be screened either directly or by artificial biosensors, with low-throughput targeted validation of the molecule of interest to uncover nonintuitive beneficial metabolic engineering targets and combinations hereof. Using this workflow, we identified several nonobvious novel targets for improving p-coumaric acid (p-CA) and L -DOPA production from two large 4k gRNA libraries each deregulating 1000 metabolic genes in the yeast Saccharomyces cerevisiae. We initially screened yeast cells transformed with gRNA library plasmids for individual regulatory targets improving the production of l-tyrosine-derived betaxanthins, identifying 30 targets that increased intracellular betaxanthin content 3.5–5.7 fold. Hereafter, we screened the targets individually in a high-producing p-CA strain, narrowing down the targets to six that increased the secreted titer by up to 15%. To investigate whether any of the six targets could be additively combined to improve p-CA production further, we created a gRNA multiplexing library and subjected it to our proposed coupled workflow. The combination of regulating PYC1 and NTH2 simultaneously resulted in the highest (threefold) improvement of the betaxanthin content, and an additive trend was also observed in the p-CA strain. Lastly, we tested the initial 30 targets in a L -DOPA producing strain, identifying 10 targets that increased the secreted titer by up to 89%, further validating our screening by proxy workflow. This coupled approach is useful for strain development in the absence of direct HTP screening assays for products of interest.

59 BASIC BIOLOGICAL SCIENCES↗