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At least 1,405 records · Page 78

Unsteady Adjoint of Pressure Loss for a Fundamental Transonic Turbine Vane

High-fidelity simulations, e.g., large eddy simulation (LES), are often needed for accurately predicting pressure losses due to wake mixing and boundary layer development in turbomachinery applications. An unsteady adjoint of high-fidelity simulations is useful for design optimization in such aerodynamic applications. In this paper, we present unsteady adjoint solutions using a large eddy simulation model for an inlet guide vane from von Karman Institute (VKI) using aerothermal objectives. The unsteady adjoint method is effective in capturing the gradient for a short time interval aerothermal objective, whereas the method provides diverging gradients for long time-averaged thermal objectives. As the boundary layer on the suction side near the trailing edge of the vane is turbulent, it poses a challenge for the adjoint solver. The chaotic dynamics cause the adjoint solution to diverge exponentially from the trailing edge region when solved backward in time. This results in the corruption of the sensitivities obtained from the adjoint solutions. An energy analysis of the unsteady compressible Navier–Stokes adjoint equations indicates that adding artificial viscosity to the adjoint equations can dissipate the adjoint energy while potentially maintaining the accuracy of the adjoint sensitivities. Analyzing the growth term of the adjoint energy provides a metric for identifying the regions in the flow where the adjoint term is diverging. Results for the vane obtained from simulations performed on the Titan supercomputer are demonstrated.

Engineering↗

Experimental Characterization and Numerical Modeling of the Interaction Between Carbon Fiber Composite Prepregs During a Preforming Process

We report carbon fiber reinforced composites have received growing attention because of their superior performance and high potential for lightweight systems. An economic method to manufacture the parts made of these composites is a sequence of forming followed by a compression molding. The first step in this sequence is called preforming that forms the prepreg, which is the fabric impregnated with the uncured resin, to the product geometry, while the molding process cures the resin. Slip between different prepreg layers is observed in the preforming step, and it is believed to have a non-negligible impact on the resulting geometry. This paper reports a method to characterize the interaction between different prepreg layers, which should be valuable for future predictive modeling and design optimization. An experimental device was built to evaluate the interactions with respect to various industrial production conditions. The experimental results were analyzed for an in-depth understanding about how temperature, relative sliding speed, and fiber orientation affect the tangential interaction between two prepreg layers. Moreover, a hydro-lubricant model was introduced to study the relative motion mechanism of this fabric-resin-fabric system, and the results agreed well with the experiment data. Lastly, the interaction factors obtained from this research will be implemented in a preforming process finite element simulation model.

36 MATERIALS SCIENCE↗

Real-Time Ecodriving Control in Electrified Connected and Autonomous Vehicles Using Approximate Dynamic Programing

Connected and automated vehicles (CAVs), particularly those with a hybrid electric powertrain, have the potential to significantly improve vehicle energy savings in real-world driving conditions. In particular, the ecodriving problem seeks to design optimal speed and power usage profiles based on available information from connectivity and advanced mapping features to minimize the fuel consumption over an itinerary. This paper presents a hierarchical multilayer model predictive control (MPC) approach for improving the fuel economy of a 48 V mild-hybrid powertrain in a connected vehicle environment. Approximate dynamic programing (DP) is used to solve the receding horizon optimal control problem, whose terminal cost is approximated with the base policy obtained from the long-term optimization. The controller was tested virtually (with deterministic and Monte Carlo simulation) across multiple real-world routes, demonstrating energy savings of more than 20%. The controller was then deployed on a test vehicle equipped with a rapid prototyping embedded controller. In-vehicle testing confirm the energy savings obtained in simulation and demonstrate the real-time ability of the controller.

Automation & Control Systems↗

Computation of Direct Sensitivities of Spatial Multibody Systems With Joint Friction

Abstract Friction exists in most mechanical systems and may have a major influence on the dynamic performance of the system. The incorporation of friction in dynamic systems has been a subject of active research for several years owing to its high nonlinearity and its dependence on several parameters. Consequently, optimization of dynamic systems with friction becomes a challenging task. Gradient-based optimization of dynamical systems is a prominent technique for optimal design and requires the computation of model sensitivities with respect to the design parameters. The novel contribution of this paper is the derivation of the analytical methodology for the computation of direct sensitivities for smooth multibody systems with joint friction using the Lagrangian index-1 formulation. System dynamics have been computed using two different friction models; the Brown and McPhee, and the Gonthier et al. model. The methodology proposed to obtain model sensitivities has also been validated using the complex finite difference method. A case study has been conducted on a spatial multibody system to observe the effect of friction on the dynamics and model sensitivities, compare sensitivities with respect to different parameters and demonstrate the numerical and validation aspects. Since design parameters can have very different magnitudes and units, the sensitivities have been scaled with the parameters for comparison. Finally, a discussion has been presented on the interpretation of the case study results. Due to the incorporation of joint friction, ‘jumps’ or discontinuities are observed in the model sensitivities akin to those observed for hybrid dynamical systems.

Engineering↗

Bayesian, Multifidelity Operator Learning for Complex Engineering Systems–A Position Paper

Abstract Deep learning has significantly improved the state-of-the-art in computer vision and natural language processing, and holds great potential to design effective tools for predicting and simulating complex engineering systems. In particular, scientific machine learning seeks to apply the power of deep learning to scientific and engineering tasks, with operator learning (OL) emerging as a particularly effective tool. OL can approximate nonlinear operators arising in complex engineering systems, making it useful for simulating, designing, and controlling those systems. In this position paper, we provide a comprehensive overview of OL, including its potential applications to complex engineering domains. We cover three variations of OL approaches: deterministic OL for modeling nonautonomous systems, OL with uncertainty quantification (UQ) capabilities, and multifidelity OL. For each variation, we discuss drawbacks and potential applications to engineering, in addition to providing a detailed explanation. We also highlight how multifidelity OL approaches with UQ capabilities can be used to design, optimize, and control engineering systems. Finally, we outline some potential challenges for OL within the engineering domain.

Computer Science↗

Effect of Wettability Gradient on the Scale Formation in Falling Film Flow

Mitigation of scale formation and performance degradation remains a vital challenge for falling film evaporators in various industries. In this work, an experimental study of falling film flow on a horizontal tube is conducted to investigate the effects of wettability gradients on thermal, hydraulic, and fouling behavior. It is revealed that certain hydrophobic coating patterns, such as strip, ring, and grid patterns, lead to unwetted heat transfer area, which results in decreased heat transfer compared to fully wetted plain tube. By adjusting the geometry and position of the wettability gradient, the hybrid coating demonstrates improved heat transfer performance. Based on the characteristics of horizontal tube falling film flow, impinging jet, thin film flow, and liquid retention at the tube bottom, a hybrid coating pattern is developed to improve surface wetting and mitigate the scaling coverage. It is revealed that scale deposition is regulated by wettability gradient. Crystals tend to be dense and compact in hydrophilic areas, while they appear scattered or even absent in hydrophobic regions, depending on the dimension of the hydrophobic area. While at the hydrophilic/hydrophobic boundary, a noticeable scale thickness step is observed, which raises the potential for self-cleaning. Furthermore, the balance of minimization of scaling layer coverage and maximization of wetting area requires an optimal design in coating dimensions, for which a systemic study of both flow dynamics and fouling characteristics on the falling film is necessary in the future.

42 ENGINEERING↗

Experimental Characterization Of The Interaction Between Carbon Fiber Composite Prepregs During The Preforming Process

Carbon fiber composites have received growing attention because of their high performance. One economic method to manufacturing the composite parts is the sequence of forming followed by the compression molding process. In this sequence, the preforming procedure forms the prepreg, which is the composite with the uncured resin, to the product geometry while the molding process cures the resin. Slip between different prepreg layers is observed in the preforming step and this paper reports a method to characterize the properties of the interaction between different prepreg layers, which is critical to predictive modeling and design optimization. An experimental setup was established to evaluate the interactions at various industrial production conditions. The experimental results were analyzed for an in-depth understanding about how the temperature, the relative sliding speed, and the fiber orientation affect the tangential interaction between two prepreg layers. The interaction factors measured from these experiments will be implemented in the computational preforming program.

Composite, Preforming, Material characterization, ↗

Design Considerations of a Slit Diaphragm Flexure Used in a Precision Mirror Gimbal

Two precision mirror gimbals were designed using slit diaphragm flexures to provide two-axis precision mirror alignment in space-limited applications. Both gimbals are currently in use in diagnostics at the National Ignition Facility: one design in the Gamma Reaction History (GRH) diagnostic and the other in the Neutron Imaging System (NIS) diagnostic. The GRH gimbal has an adjustment sensitivity of 0.1 mrad about both axes and a total adjustment capability of ±6°; the NIS gimbal has an adjustment sensitivity of 0.8 μrad about both axes and a total adjustment range of ±3°. Both slit diaphragm flexures were electro-discharge machined out of high-strength titanium and utilize stainless steel stiffeners. The stiffener-flexure design results in adjustment axes with excellent orthogonality and centering with respect to the mirror in a single stage; a typical two-axis gimbal flexure requires two stages. Finite element analyses are presented for both flexure designs, and a design optimization of the GRH flexure is discussed.

42 ENGINEERING↗

A review of breast tomosynthesis. Part I. The image acquisition process

Mammography is a very well-established imaging modality for the early detection and diagnosis of breast cancer. However, since the introduction of digital imaging to the realm of radiology, more advanced, and especially tomographic imaging methods have been made possible. One of these methods, breast tomosynthesis, has finally been introduced to the clinic for routine everyday use, with potential to in the future replace mammography for screening for breast cancer. In this two part paper, the extensive research performed during the development of breast tomosynthesis is reviewed, with a focus on the research addressing the medical physics aspects of this imaging modality. This first paper will review the research performed on the issues relevant to the image acquisition process, including system design, optimization of geometry and technique, x-ray scatter, and radiation dose. The companion to this paper will review all other aspects of breast tomosynthesis imaging, including the reconstruction process.

60 APPLIED LIFE SCIENCES↗

Evaluation of the microangiographic fluoroscope (MAF) using generalized system performance metrics

Purpose: The performance of a newly developed, high resolution, microangiographic fluoroscope (MAF) (35 {mu}m pixel pitch and 300 {mu}m thick CsI phosphor) was evaluated using a generalized linear system analysis and compared with that of a standard amorphous Si thin film transistor flat panel detector (FPD) (194 {mu}m pixel pitch and 600 {mu}m thick CsI phosphor). The linear system metrics such as modulation transfer function (MTF), noise power spectrum, and detection quantum efficiency (DQE) are commonly used to gauge the intrinsic detector performance in the detector plane. However, these linear system metrics do not provide information about the image receptor performance in a real system since they do not include the effects of other parameters such as focal spot distribution, scatter radiation, and geometric unsharpness, which may compromise detector performance characteristics. Use of generalized linear system metrics [generalized modulation transfer function (GMTF), generalized normalized noise power spectrum (GNNPS), and generalized detection quantum efficiency (GDQE)] that include these effects gives a more meaningful, complete, and appropriate evaluation of detector performance as part of the imaging system. Methods: A uniform head equivalent phantom was used to simulate realistic clinical parameters and x-ray spectra. The detector MTFs were measured using the slanted edge method and the focal spot MTFs were measured using a pinhole assembly. The scatter MTF was simulated and the scatter fraction was measured for a head-equivalent phantom. The generalized system metrics were calculated for different combinations of three choices of focal spots and three different magnifications with two different air-gaps. The performance of the MAF was also illustrated using stent images obtained with different focal spots under similar conditions. Results: Results for the generalized metrics provide a quantitative description of the performance of the imaging system for both detectors. This generalized analysis demonstrated that both detectors have similar imaging capabilities at lower spatial frequencies, but that the MAF has superior performance over the FPD at higher frequencies even when considering focal spot blurring and scatter. Conclusions: This generalized performance analysis demonstrates the significance of focal spot size, magnification, and scatter on the system performance metrics (GMTF, GNNPS, and GDQE). Although the ideal detector performance characteristics of the MAF are not fully realized due to these other system factors, it still retains an advantage in DQE at high spatial frequencies over the FPD. Similar studies based on the generalized linear system metrics can serve as an efficient tool to evaluate total system capabilities under different realistic conditions to enable optimal design for specific imaging tasks.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

SU-E-T-53: Benchmarking a Monte Carlo Model for Patient Plane Leakage Calculations of Low Energy 6MV Unique Linacs

Purpose: To validate the physics parameters of a Monte Carlo model for patient plane leakage calculations on the 6MV Unique linac by comparing the simulations against IEC patient plane leakage measurements. The benchmarked model can further be used for shielding design optimization, to predict leakage in the proximity of intended treatment fields, reduce the system weight and cost, and improve components reliability. Methods: The treatment head geometry of the Unique linac was simulated in Geant4 (v9.4.p02 with “Opt3” standard electromagnetic physics list) based on CAD drawings of all collimation and shielding components projected from the target to the area within 2m from isocenter. A 4×4m2 scorer was inserted 1m from the target in the patient plane and multiple phase space files were recorded by performing a 40-node computing cluster simulation on the EC2 cloud. The photon energy fluence was calculated relative to the value at isocenter for a 10×10cm2 field using 10×10mm2 bins. Tungsten blocks were parked accordingly to represent MLC120. The secondary particle contamination to patient plane was eliminated by “killing” those particles prior to the primary collimator entrance using a “kill-plane”, which represented the upper head shielding components not being modeled. Both IEC patient-plane leakage and X/Y-jaws transmission were simulated. Results: The contribution of photons to energy fluence was 0.064% on average, in excellent agreement with the experimental data available at 0.5, 1.0, and 1.5m from isocenter, characterized by an average leakage of 0.045% and a maximum leakage of 0.085%. X- and Y-jaws transmissions of 0.43% and 0.44% were found in good agreement with measurements of 0.48% and 0.43%, respectively. Conclusion: A Geant4 model based on energy fluence calculations for the 6MV Unique linac was created and validated using IEC patient plane leakage measurements. The “kill-plane” has effectively eliminated electron contamination to patient plane in these simulations.

60 APPLIED LIFE SCIENCES↗

SU-E-T-512: Electromagnetic Simulations of the Dielectric Wall Accelerator

Purpose: To characterize and parametrically study the key components of a dielectric wall accelerator through electromagnetic modeling and particle tracking. Methods: Electromagnetic and particle tracking simulations were performed using a commercial code (CST Microwave Studio, CST Inc.) utilizing the finite integration technique. A dielectric wall accelerator consists of a series of stacked transmission lines sequentially fired in synchrony with an ion pulse. Numerous properties of the stacked transmission lines, including geometric, material, and electronic properties, were analyzed and varied in order to assess their impact on the transverse and axial electric fields. Additionally, stacks of transmission lines were simulated in order to quantify the parasitic effect observed in closely packed lines. Particle tracking simulations using the particle-in-cell method were performed on the various stacks to determine the impact of the above properties on the resultant phase space of the ions. Results: Examination of the simulation results show that novel geometries can shape the accelerating pulse in order to reduce the energy spread and increase the average energy of accelerated ions. Parasitic effects were quantified for various geometries and found to vary with distance from the end of the transmission line and along the beam axis. An optimal arrival time of an ion pulse relative to the triggering of the transmission lines for a given geometry was determined through parametric study. Benchmark simulations of single transmission lines agree well with published experimental results. Conclusion: This work characterized the behavior of the transmission lines used in a dielectric wall accelerator and used this information to improve them in novel ways. Utilizing novel geometries, we were able to improve the accelerating gradient and phase space of the accelerated particle bunch. Through simulation, we were able to discover and optimize design issues with the device at low cost. Funding: Morgridge Institute for Research, Madison WI; Conflict of Interest: Dr. Mackie is an investor and board member at CPAC, a company developing compact accelerator designs similar to those discussed in this work, but designs discussed are not directed by CPAC. Funding: Morgridge Institute for Research, Madison WI; Conflict of Interest: Dr. Mackie is an investor and board member at CPAC, a company developing compact accelerator designs similar to those discussed in this work, but designs discussed are not directed by CPAC.

43 PARTICLE ACCELERATORS↗

SU-F-18C-05: Characterization of a Silicon Strip Photon-Counting Detector in the Presence of Compton Scatter: A Simulation Study

Purpose: To investigate the effect of Compton scatter on detection efficiency and charge-sharing for a Si strip photon-counting detector as a function of pixel pitch, slice thickness and total pixel length. Methods: A CT imaging system employing a silicon photon-counting detector was implemented using the GATE Monte Carlo package. A focal spot size of 300 µm, magnification of 1.33, and pixel pitches of 0.1 and 0.5mm were initially investigated. A 60 kVp spectrum with 3 mm Al filter was used and energy spectral degradation based on a prototype detector was simulated. To study charge-sharing, a single pixel was illuminated, and the detector response in neighboring pixels was investigated. A longitudinally semiinfinite detector was simulated to optimize the quantum detection efficiency of the imaging system as a function of pixel pitch, slice thickness and depth of interaction. A 2.5 mm thick tungsten plate with a 0.01 mm by 1.5 mm slit was implemented to calculate the modulation transfer function (MTF) from projection-based images. A threshold of 15 keV was implemented in the detector simulation. The preliminary charge sharing investigation results considered only scattering effects and the detector electronics related factors were neglected. Results: Using a 15 keV threshold, 1% of the pixel charge migrated into neighboring pixels with a pixel size of 0.1×0.1 mm{sup 2}. The quantum detection efficiency was 77%, 84%, 87% and 89% for 15 mm, 22.5 mm, 30 mm, and 45 mm length silicon detector pixels, respectively. For a pixel pitch of 0.1 mm, the spatial frequency at 10% of the maximum MTF was found to be 5.2 lp/mm. This agreed with an experimental MTF measurement of 5.3 lp/mm with a similar detector configuration. Conclusion: Using optimized design parameters, Si strip photon-counting detectors can offer high detection efficiency and spatial resolution even in the presence of Compton scatter.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Feasibility study of a dual detector configuration concept for simultaneous megavoltage imaging and dose verification in radiotherapy

Purpose: To test the feasibility of a dual detector concept for comprehensive verification of external beam radiotherapy. Specifically, the authors test the hypothesis that a portal imaging device coupled to a 2D dosimeter provides a system capable of simultaneous imaging and dose verification, and that the presence of each device does not significantly detract from the performance of the other. Methods: The dual detector configuration comprised of a standard radiotherapy electronic portal imaging device (EPID) positioned directly on top of an ionization-chamber array (ICA) with 2 cm solid water buildup material (between EPID and ICA) and 5 cm solid backscatter material. The dose response characteristics of the ICA and the imaging performance of the EPID in the dual detector configuration were compared to the performance in their respective reference clinical configurations. The reference clinical configurations were 6 cm solid water buildup material, an ICA, and 5 cm solid water backscatter material as the reference dosimetry configuration, and an EPID with no additional buildup or solid backscatter material as the reference imaging configuration. The dose response of the ICA was evaluated by measuring the detector’s response with respect to off-axis position, field size, and transit object thickness. Clinical dosimetry performance was evaluated by measuring a range of clinical intensity-modulated radiation therapy (IMRT) beams in transit and nontransit geometries. The imaging performance of the EPID was evaluated quantitatively by measuring the contrast-to-noise ratio (CNR) and spatial resolution. Images of an anthropomorphic phantom were also used for qualitative assessment. Results: The measured off-axis and field size response with the ICA in both transit and nontransit geometries for both dual detector configuration and reference dosimetry configuration agreed to within 1%. Transit dose response as a function of object thickness agreed to within 0.5%. All IMRT test patterns and clinical IMRT beams had gamma pass rates of ≥98% at 2%/2 mm criteria. In terms of imaging performance, the measured CNR and spatial resolution (f{sub 50}) were 263.23 ± 24.85 and 0.4025 ± 1.25 × 10{sup −3} for dual detector configuration and 324 ± 26.65 and 0.4141 ± 1.14 × 10{sup −3} for reference imaging configuration, respectively. The CNR and spatial resolution were quantitatively worse in the dual detector configuration due to the additional backscatter. The difference in imaging performance was not visible in qualitative assessment of phantom images. Conclusions: Combining a commercially available ICA dosimetry device with a conventional EPID did not significantly detract from the performance of either device. Further improvements in imaging performance may be achieved with an optimized design. This study demonstrates the feasibility of a dual detector concept for simultaneous imaging and dosimetry in radiation therapy.

60 APPLIED LIFE SCIENCES↗

A self-powered thin-film radiation detector using intrinsic high-energy current

Purpose: The authors introduce a radiation detection method that relies on high-energy current (HEC) formed by secondary charged particles in the detector material, which induces conduction current in an external readout circuit. Direct energy conversion of the incident radiation powers the signal formation without the need for external bias voltage or amplification. The detector the authors consider is a thin-film multilayer device, composed of alternating disparate electrically conductive and insulating layers. The optimal design of HEC detectors consists of microscopic or nanoscopic structures. Methods: Theoretical and computational developments are presented to illustrate the salient properties of the HEC detector and to demonstrate its feasibility. In this work, the authors examine single-sandwiched and periodic layers of Cu and Al, and Au and Al, ranging in thickness from 100 nm to 300 μm and separated by similarly sized dielectric gaps, exposed to 120 kVp x-ray beam (half-value thickness of 4.1 mm of Al). The energy deposition characteristics and the high-energy current were determined using radiation transport computations. Results: The authors found that in a dual-layer configuration, the signal is in the measurable range. For a defined total detector thickness in a multilayer structure, the signal sharply increases with decreasing thickness of the high-Z conductive layers. This paper focuses on the computational results while a companion paper reports the experimental findings. Conclusions: Significant advantages of the device are that it does not require external power supply and amplification to create a measurable signal; it can be made in any size and geometry, including very thin (sub-millimeter to submicron) flexible curvilinear forms, and it is inexpensive. Potential applications include medical dosimetry (both in vivo and external), radiation protection, and other settings where one or more of the above qualities are desired.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Sci-Sat AM: Radiation Dosimetry and Practical Therapy Solutions - 11: Commissioning of a system for the measurement of electron stopping powers

Purpose: To develop the techniques required to experimentally determine electron stopping powers for application in primary standards and dosimetry protocols. Method and Materials: A large-volume HPGe detector system (>80% efficiency) was commissioned for the measurement of high energy (5–35 MeV) electron beams. As a proof of principle the system was used with a Y-90/Sr-90 radioactive source. Thin plates of absorbing material (< 0.1 gcm-2) were then placed between the source and detector and the emerging electron spectrum was acquired. The full experimental geometry was modelled using the EGSnrc package to validate the detector design, optimize the experimental setup and compare measured and calculated spectra. Results: The biggest challenge using a beta source was to identify a robust spectral parameter to determine for each measurement. An end-point-fitting routine was used to determine the maximum energy, Emax, of the beta spectrum for each absorber thickness t. The parameter dEmax/dt is related to the electron stopping power and the same routine was applied to both measured and simulated spectra. Although the standard uncertainty in dEmax/dt was of the order of 5 %, by taking the ratio of measured and Monte Carlo values for dEmax/dt the uncertainty of the fitting routine was eliminated and the uncertainty was reduced to less than 2 %. The agreement between measurement and simulation was within this uncertainty estimate. Conclusion: The investigation confirmed the experimental approach and demonstrated that EGSnrc could accurately determine correction factors that will be required for the final measurement setup in a linac beam.

61 RADIATION PROTECTION AND DOSIMETRY↗

Activity profiling and crystal structures of inhibitor-bound SARS-CoV-2 papain-like protease: A framework for anti–COVID-19 drug design

Viral papain-like cysteine protease (PLpro, NSP3) is essential for SARS-CoV-2 replication and represents a promising target for the development of antiviral drugs. Here, we used a combinatorial substrate library and performed comprehensive activity profiling of SARS-CoV-2 PLpro. On the scaffold of the best hits from positional scanning, we designed optimal fluorogenic substrates and irreversible inhibitors with a high degree of selectivity for SARS PLpro. We determined crystal structures of two of these inhibitors in complex with SARS-CoV-2 PLpro that reveals their inhibitory mechanisms and provides a molecular basis for the observed substrate specificity profiles. Last, we demonstrate that SARS-CoV-2 PLpro harbors deISGylating activity similar to SARSCoV-1 PLpro but its ability to hydrolyze K48-linked Ub chains is diminished, which our sequence and structure analysis provides a basis for. Together, this work has revealed the molecular rules governing PLpro substrate specificity and provides a framework for development of inhibitors with potential therapeutic value or drug repurposing.

59 BASIC BIOLOGICAL SCIENCES↗

Multiplexed CRISPR-Cas9-Based Genome Editing of Rhodosporidium toruloides

Microbial production of biofuels and bioproducts offers a sustainable and economic alternative to petroleum-based fuels and chemicals. The basidiomycete yeast Rhodosporidium toruloides is a promising platform organism for generating bioproducts due to its ability to consume a broad spectrum of carbon sources (including those derived from lignocellulosic biomass) and to naturally accumulate high levels of lipids and carotenoids, two biosynthetic pathways that can be leveraged to produce a wide range of bioproducts. While R. toruloides has great potential, it has a more limited set of tools for genetic engineering relative to more advanced yeast platform organisms such as Yarrowia lipolytica and Saccharomyces cerevisiae. Significant advancements in the past few years have bolstered R. toruloides ’ engineering capacity. Here we expand this capacity by demonstrating the first use of CRISPR-Cas9-based gene disruption in R. toruloides . Transforming a Cas9 expression cassette harboring nourseothricin resistance and selecting transformants on this antibiotic resulted in strains of R. toruloides exhibiting successful targeted disruption of the native URA3 gene. While editing efficiencies were initially low (0.002%), optimization of the cassette increased efficiencies 364-fold (to 0.6%). Applying these optimized design conditions enabled disruption of another native gene involved in carotenoid biosynthesis, CAR2 , with much greater success; editing efficiencies of CAR2 deletion reached roughly 50%. Finally, we demonstrated efficient multiplexed genome editing by disrupting both CAR2 and URA3 in a single transformation. Altogether, our results provide a framework for applying CRISPR-Cas9 to R. toruloides that will facilitate rapid and high-throughput genome engineering in this industrially relevant organism.

59 BASIC BIOLOGICAL SCIENCES↗