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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 145 records · Page 8
All sourcesNASA NTRSDOE OSTI
DOE OSTI2025

EVI-LOCATE User Manual

One of the longest stages in the deployment of electric vehicle supply equipment (EVSE) is the initial planning of the infrastructure itself. Engineers and fleet experts from the National Renewable Energy Laboratory (NREL) have supported dozens of charging infrastructure site plans over the past couple decades, including the generation of site schematics, determinations of electric capacity, and estimates for likely costs. As the market for electric vehicles (EVs) has matured, this approach should no longer require a time and personnel intensive process. In order to shorten the time taken to develop site plans and cost estimates, NREL developed a tool that fleet managers, facility managers, electricians, EVSE installers, and members of the public can use to develop initial schematics and ballpark pricing for charging station installations. The Electric Vehicle Infrastructure - Locally Optimized Charger Assessment Tool and Estimator (EVI-LOCATE) provides a structured and consistent way for users to enter information about their planned EVSE project in a relatively simple web-based format. EVI-LOCATE then calculates electrical equipment capacity, wiring runs, and project costs. It produces a site diagram optimized around surface characteristics with differential trenching costs for softscape such as grass compared to hardscape such as asphalt that can be adjusted by users in the tool. It also stores the resulting site plans and costs in a dashboard for access at a later date, including plan revisions if necessary. This document guides users through the EVI-LOCATE screens and associated questions. It contains tip text boxes throughout on how best to interface with the tool and find additional information or context. The appendices contain the assumptions and calculations underpinning the tool. Much of the information for EVI-LOCATE was gathered through industry engagements with EVSE installers, invoices from completed EVSE installations, Gordian's RS Means construction data, and the General Services Administration blanket purchase agreement for EVSE. For a visual tutorial of the tool, users can watch EVI-LOCATE Step-by-Step Video. The tool itself is available at https://evi-locate.nrel.gov.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗
DOE OSTI2021

ESnet Secure Copy (EScp) v0.6

EScp is a high speed transfer tool with a similar command line syntax to scp. Unlike SCP it is designed to transfer files at high speed, thus far we have been able to show 100gbit/s transfers, although I expect that the throughput should scale in proportion to the network interface, i.e. I expect 400gbit/s performance on our 400gbit/s test bed. EScp achieves good performance through an innovative design (multithreaded, zero copy transfers), along with pluggable filters and I/O engines. As an example, you can switch from POSIX i/O to UIO by checking a different engine. It also natively supports encryption, and cheksums for file verification and transport security. AAA is through standard SSH (same as SCP). By taking advantage of filters, EScp supports transferring unstructured data and/or I/O to non-posix data sources. Examples include streaming data (i.e. from equipment), transferring data to the cloud, and/or supporting non-posix file systems (like HPSS).

Shiflett, Charles↗
DOE OSTI2021

Securing Distributed Energy Resource Integration

The penetration of distributed energy resources (DER) is growing at much higher rates than predicted 20 years ago. Far from being used only in residential settings, DER are now installed on distribution and transmission circuits. In this position, they do not have the same properties as traditional generators and are more flexible in many cases. The growing penetration and range of uses for DER motivate the need to reliably and safely integrate them into the grid. Operators must be able to rely on them not only for normal operation, but also during abnormal conditions like black starts or adverse cyber scenarios. To that end, we study the communications, device interfaces, and potential consequences of DER operation under abnormal and adversarial conditions. The weaknesses of communications networks are studied based on the industrial protocols used, and the benefits of security features are examined. The device interfaces are found to be vulnerable to attack based on the requirements in the IEEE-1547 standard for DER interconnection and interoperability, which is expected to be adopted in the next ten years. In addition to exploring the requirements of the standard, we show that these vulnerabilities and others do exist and can be used maliciously in a modern storage system DER. Consequences of these vulnerabilities range from exacerbated grid instability, to simultaneous loss of large portions of DER penetration, to physical damage to inverters or DER themselves and other sensitive equipment. We tie these outcomes to specific attacker actions in an effort to give operators a better threat intelligence view that allows them to prioritize mitigations. Finally, we discuss mitigations that could prevent many of the adversarial scenarios described. Some solutions can be added to existing infrastructure, while others may require longer term planning for grid modernization with consideration for security.

25 ENERGY STORAGE↗
DOE OSTI2021

Architecture Of A Multi-channel Data Streaming Device With An Fpga As A Coprocessor

The design of a data acquisition system often involves the integration of a Field Programmable Gate Array (FPGA) with analog front-end components to achieve precise timing and control. Reuse of these hardware systems can be difficult since they need to be tightly coupled to the communications interface and timing requirements of the specific ADC used. A hybrid design exploring the use of FPGA as a coprocessor to a traditional CPU in a dataflow architecture is presented. Reduction in the volume of data and gradual transitioning of data processing away from a hard real-time environment are both discussed. Chief design concerns, including data throughput and precise synchronization with external stimuli, are addressed. The discussion is illustrated by the implementation of a multi-channel digital integrator, a device based entirely on commercial off-the-shelf (COTS) equipment.

Nogiec, Jerzy M.↗
DOE OSTI2025

Quantifying Twist Angles in Cuprate Heterostructures with Anisotropic Raman Signatures

Artificially engineered twisted van der Waals (vdW) heterostructures have unlocked new pathways for exploring emergent quantum phenomena and strongly correlated electronic states. Many of these phenomena are highly sensitive to the twist angle, which can be deliberately tuned to tailor the interlayer interactions. This makes the twist angle a critical tunable parameter, emphasizing the need for precise control and accurate characterization during device fabrication. In particular, twisted cuprate heterostructures based on Bi 2 Sr 2 CaCu 2 O 8 + x (BSCCO) have demonstrated angle-dependent superconducting properties, positioning the twist angle as a key tunable parameter. However, the twisted interface is highly unstable under ambient conditions and vulnerable to damage from conventional characterization tools such as electron microscopy or scanning probe techniques. In this work, a fully non-invasive, polarization-resolved Raman spectroscopy approach is introduced for determining twist angles in artificially stacked BSCCO heterostructures. By analyzing twist-dependent anisotropic vibrational Raman modes, particularly utilizing the out-of-plane A 1g vibrational mode of Bi/Sr at ≈116 cm −1 , clear optical fingerprints of the rotational misalignment between cuprate layers are identified. The high-resolution confocal Raman setup, equipped with polarization control and RayShield filtering down to 10 cm −1 , allows for reliable and reproducible measurements without compromising the material's structural integrity.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗
DOE OSTI2024

Operando XPS for Plasma Process Monitoring: A Case Study on the Hydrogenation of Copper Oxide Confined under h-BN

Here, we demonstrate that ambient pressure x-ray photoelectron spectroscopy (APXPS) can be used for in situ studies of dynamic changes in surface chemistry in a plasma environment. This opens a new and vast application space for XPS and greatly complements modern spectroscopy techniques to probe plasma-solid/liquid interactions relevant to process monitoring in the semiconductor industry, bio-medical plasma applications and plasma remediation technologies. Hexagonal boron nitride (h-BN) grown on Cu was used in this study as a well-defined model system for plasma process monitoring and because of its unique chemical, optical and electrical properties that make it a prospective material for advanced electronics. To better understand the stability and surface chemistry of h-BN during plasma assisted processing, we track in real time the plasma-induced chemical state changes of B, N and the underlying Cu substrate using APXPS equipped with an AC discharge plasma source operating at 13 Pa. Residual gas analysis (RGA) mass-spectra were concurrently collected during plasma-XPS to track reaction products formed during plasma exposure. A clear reduction of Cu x O is seen, while an h-BN layer remains intact, suggesting hydrogen radical (H • ) species can attack the exposed and h-BN covered Cu oxide patches and partially reduce the underlying substrate without significantly damaging the overlaying h-BN, which is of practical importance for development of h-BN encapsulated devices and interfaces. In addition to demonstration of plasma-XPS capabilities we discuss the observed challenges (e.g., parasitic plasma-chamber walls reactions and charging effects) and propose potential solutions.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2026

Evi-Pro Lite API

This application programing interface provides output from NLR's EVI-Pro model and is used to power the EVI-Pro Lite tool at https://afdc.energy.gov/evi-pro-lite. These endpoints provide daily (24-hour) fleet-level charging load profiles for a variety of customizable scenarios.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗
DOE OSTI2025

Hands-On, Heads-Up: Blending Cyber T&E with Data Science-Driven Training in Jupyter Notebooks

In an era of increasingly sophisticated threats to critical infrastructure, cybersecurity professionals must be more than just aware; they must be immersed, agile, and equipped to operate in environments where failure is not an option. Nowhere is this truer than in the nuclear sector, where cyber-physical systems, regulatory scrutiny, and insider threat potential demand a new generation of hands-on, technically fluent defenders. This paper presents a unified training approach that integrates Cybersecurity Test and Evaluation (T&E) with data science techniques using Jupyter Notebooks as the interactive lab environment. The program centers on a modular, scenario-driven curriculum designed to build not just knowledge but practical capability in the assessment and defense of radiation detection systems, firmware interfaces, and operational security postures.

98 - NUCLEAR DISARMAMENT, SAFEGUARDS, AND PHYSICAL↗
DOE OSTI2026

ORCHID: Orchestrated Retrieval-Augmented Classification of High-Risk Property with Intelligent Decision-Making

High-Risk Property (HRP) classification is critical at U.S. Department of Energy (DOE) sites, where inventories include sensitive and often dual-use equipment. Compliance must track evolving rules designated by various export control policies to make transparent and auditable decisions. Traditional expert-only workflows are time-consuming, backlog-prone, and struggle to keep pace with shifting regulatory boundaries. We propose ORCHID, a modular agentic framework for HRP classification that pairs retrieval-augmented generation (RAG) with human oversight to produce policy based outputs that can be audited. Small cooperating agents—retrieval, description refiner, classifier, validator, and feedback logger—coordinate via agent-to-agent messaging and invoke tools through the Model Context Protocol (MCP) for model-agnostic on-premise operation. The interface follows an "Item to Evidence to Decision" loop with step-by-step reasoning, on-policy citations, and append-only audit bundles (run-cards, prompts, evidence). In preliminary tests on real HRP cases, ORCHID improves accuracy and traceability over a non-agentic baseline while deferring uncertain items to Subject Matter Experts (SMEs). The demonstration shows single item submission, grounded citations, SME feedback capture, and exportable audit artifacts—illustrating a practical path to trustworthy LLM assistance in sensitive DOE compliance workflows.

Das, Sanjay [ORNL] (ORCID:0009000542591915)↗
DOE OSTI2021

Impact of Thermal Stress on Attachment and Stability of High Temperature Strain Sensors

Static and dynamic strain sensing is required in high-temperature (HT), harsh-environments (HE) for industrial, aerospace, and energy sector applications to ensure equipment and process safety, reduce the cost of operation and maintenance, and increase process efficiency. Challenges that arise in HT HE sensing applications include device mounting, packaging, integrity, and stability to HT HE conditions. In previously reported work, static and dynamic strain surface acoustic wave resonator (SAWR) sensors were fabricated on langasite (LGS) and mounted on Inconel 625 strain beams for wireless testing up to 400°C. In this work, it has been identified that after subjecting the mounted SAWR strain sensor to thermal cycling between 100°C and 425°C, the measured sensitivity to dynamic strain decreased by 73% due to cracking at the adhesive/LGS interface, further deteriorating after additional thermal cycles. Strain modeling of the mounted LGS sensor chip up to 400°C revealed the existence of concentrated strain at the borders of the LGS chip. Microcracks caused by dicing make the chip boarders the most susceptible location for cracks to initiate when the sensor is subjected to thermal stress. In an attempt to mitigate the high strain at the LGS chip borders due to heating, adhesive shaping is proposed in this work. Simulations indicate a strain reduction of 50% at the border is achieved using both circular and triangular adhesive shapes, while also reducing the maximum strain over the entire adhesive/LGS interface by around 30%. Furthermore, the technique is thus promising for improving the integrity, reliability, and stability of static and dynamic strain sensors, particularly while operating under HT HE with several hundreddegree Celsius temperature excursions.

47 OTHER INSTRUMENTATION↗
DOE OSTI2021

Bridging Land-Surface Fluxes and Aerosol Concentrations to Triggering Convective Rainfall

During the period August 24-28, 2016, a delayed component the GO-Amazon Project, we launched nine altitude-controlled free balloons (CMET). Smaller than typical rawinsondes, CMET are equipped with altitude control, global communication via Iridium satellite, and aspirated sensors. The aims of our effort were to examine the interactions among convective boundary layer and dual river breeze circulations near the confluence of the Tapajos and Amazon Rivers in the eastern Basin. The week-long field campaign was timed to examine the reestablishment of the breeze circulations shortly after the passage of a strong instability line on August 22. Nine CMET were launched at the Curua-Una hydroelectric dam (2.8S; 54.3W), timed to encounter the Tapajos river breeze front by late afternoon. Soundings were made to establish the thickness of interface between the easterly trade and westerly Tapajos breeze circulation. Careful use of sounding strategies allowed these free balloons to track along the northerly channeled flow in the lowest 300 m above the River. Following the river encounter, balloons tracked to the west, sounding to describe the diurnal course of boundary layer in the forest west of the Tapajos River. The longest flight traveled more than 770 km over three days and twice rested overnight in the rain forest canopy. Ancillary data from surface climate and flux stations as well as the Santarem radiosonde, satellite images will be used to illustrate how the breeze circulations are seen near the surface and how they were disrupted by larger-scale events. Comparisons with HYSPLIT trajectories illustrate how sensitive real trajectories are to the refraction that the encounter with the breeze effects. A second effort was intensive analyses of data from the eddy flux tower at the Tapajós National Forest near Santarém Pará Brazil, with special emphasis on the role of clouds and canopy presence on light absorbance and photosynthetic activity.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2023

Multimodal CO2 Transportation Cost Model

This model provides a cost estimate for transporting CO2 via truck or rail in the United States using commercially available equipment and technologies. The model includes an analysis of direct and indirect CO2 emissions to determine costs per net tonne of CO2 transported. Publicly available data and methods published in the peer-reviewed literature are used to the extent possible; references are available at the bottom of the "Calculations" sheet. Upstream (i.e., liquefaction, buffer storage) and downstream (i.e., buffer storage, reconditioning) activities are included in the model of emissions and costs. All capital and operating expenditures are estimated in the "Calculations" sheet. The cost of financing the project is determined in the "FINEX" sheet. All user inputs are done via drop down menus in on the "User Interface" sheet. Summary results are also provided on the "User Interface" sheet.

Myers, CoreyA↗
DOE OSTI2025

Autonomous Fueling System for Heavy-Duty Fuel Cell Electric Trucks

The motivation for this project stemmed from the challenges associated with rapidly refueling heavy-duty hydrogen fuel cell electric trucks (FCETs). Current manual refueling processes for fast refueling involve large, heavy equipment (e.g., hoses three times heavier than standard) and pose ergonomic risks and potential for equipment damage. The goal was to develop and test an autonomous fueling system to improve ergonomics, enhance safety, increase equipment durability through design improvements, and potentially speed up the fueling process. This project aimed to add to the understanding of autonomous systems in the context of heavy-duty hydrogen refueling, evaluating the technical effectiveness of potential concepts. A successfully developed system would benefit the public by facilitating the adoption of zero-emission heavy-duty transport, reducing reliance on manual labor for a physically demanding task, and potentially improving the safety and efficiency of hydrogen refueling infrastructure. The major accomplishment during the project's active period was the completion of the system-level architecture task. This involved establishing a detailed list of system requirements covering interfaces, environmental conditions, regulatory compliance, industry standards, safety, security, performance capabilities, and optional features. Five key use cases for the autonomous system were also identified. However, due to internal restructuring at Nikola, the necessary resources could not be allocated to continue the project. Consequently, Nikola opted to discontinue the project. The award was mutually terminated by Nikola and the DOE.

08 HYDROGEN↗
DOE OSTI2024

Institute for Catalysis in Energy Processes (ICEP) (Final Report)

The Institute for Catalysis in Energy Processes (ICEP) was a multi-PI program located at the Northwestern University Center for Catalysis and Surface Science from 2006-2024. Over several renewal cycles and several organizing themes, ICEP addressed fundamental questions in catalysis science. In turn, the scientific questions addressed were directly relevant to the efficient use of the nation’s resources to produce fuels and commodity chemicals, harness alternate energy sources for chemical reactions such as light or electricity, reduce emissions and waste, and minimize the impact of our use of plastics. Selective oxidation, (oxidative) dehydrogenation, deNOx, CO 2 reduction, hydrogen release, polymer decomposition / depolymerization, and many other reactions of intense interest to the US Department of Energy were studied in the center. Major ICEP strengths were in catalyst synthesis, in measurements that elucidated catalyst properties, in reaction mechanisms and kinetics, and in predictive theory and modeling. ICEP researchers were often the inventors or developers of materials and methods that were brought to bear on the Institute’s catalytic systems. Key characterization tools advanced through this project included resonance Raman and related techniques, sum frequency generation, and X-ray standing wave spectroscopy. Center members relied extensively on computational tools such as density functional theory and microkinetic modeling to fully investigate catalyst structures and catalytic mechanisms. ICEP researchers were early developers of atomic layer deposition (ALD) for catalyst synthesis and were early pioneers of metal organic frameworks for catalysis. ICEP innovations are found in areas of intense research such as single-atom catalysts, catalytic deconstruction of plastics, and catalytic MOFs, to name a few. Research areas initiated by ICEP indirectly led to DOE EFRCs, startup companies, and other achievements. Over its 18 years of existence, ICEP involved two different PIs (Stair and Notestein) and 24 senior investigators across several academic disciplines at Northwestern University. There were tight collaborations with Argonne National Laboratory, including specialized equipment and experiments permanently located there. The center supported ~400 person-years of effort by graduate students and postdocs, either directly through DOE support or indirectly by leveraging independent support provided to the trainees, e.g. through the NSF GRFP or internal fellowships. The project resulted in 328 manuscripts and numerous conference presentations.

36 MATERIALS SCIENCE↗
DOE OSTI2024

Improved Creep Testing Approach for Bentonite EBS

In the United States, the nuclear reactor contributes 18.7 percent of the nation’s total electricity generation and produces a significant amount of spent nuclear fuel (SNF) and high-level waste. Because SNF is being stored for longer periods than initially envisioned, the U.S. Department of Energy Office of Nuclear Energy, Office of Spent Fuel and Waste Science and Technology is assessing the technical performance of the SNF storage systems after extended durations. The concept of a sustainable geologic repository for SNF disposal relies heavily on the safety provided by multiple barriers, such as repository host rock, overlying rock formation, and human-made engineered barrier systems (EBS). One of the major components of EBS is bentonite-based buffer materials that isolate the nuclear-waste canisters from the host rock and fill the void left in the horizontally drilled boreholes. The resaturated bentonite has a swelling characteristic, which is useful for self-sealing the microcracks in the host rock and supporting the canister’s heavy weight. However, factors such as nonuniformity in the saturation, swelling pressure of bentonite, and the high temperature at the canister-bentonite interface may lead to instability of the waste canister inside the borehole. Therefore, evaluating the long-term deformation of bentonite-based material is necessary. RESPEC Company, LLC recently completed a preliminary study for the U.S. Department of Energy (DE SC0022804) that is attempting to understand the time-dependent deformation in the consolidated sand/bentonite (SB) specimens through triaxial creep experiments and improve the understanding of EBS performance under anticipated repository conditions. The project included developing a standard testing procedure for preparing the consolidated core specimen from a mixture of sand and bentonite in a 50:50 ratio by weight, fabricating a new pressure vessel in-house, modifying existing creep equipment to perform triaxial creep experiments on consolidated SB specimens, and performing six long-term triaxial creep experiments under low-deviatoric stress at two different temperature conditions, which is representative of hypothetical conditions that might be encountered in a geologic repository within a reasonable rate of success in completing creep experiments. The consolidated SB specimen was prepared using the novel specimen preparation procedure and had physical characteristics (e.g., moisture content and bulk density) reasonably comparable with the properties of bentonite-based buffer, which is proposed to be used in the EBS in the Swedish KBS-3H design and the Swiss design of SNF disposal in the horizontal drifts. The newly fabricated pressure vessel could withstand the confining pressure of up to 15 megapascals (MPa) while maintaining the test temperature of up to 90 degrees Celsius (°C) for long-term creep experiments. The preliminary results from the triaxial creep experiments suggested that the consolidated SB specimen may experience time-dependent deformation at a low-deviatoric stress state and room temperature condition. Based on the deformation recorded in the SB specimen from axial and radial linear variable differential transformers (LVDTs) and physical characteristic changes, it is apparent that the time-dependent deformation is a combination of consolidation and creep, which is influenced by the level of deviatoric stresses and temperature. The systematic creep testing of bentonite-based specimens helped develop a standard testing procedure applicable for analyzing the similar characteristics of shale or clay-like materials in different geologic conditions outside repository science, including in civil engineering and infrastructure projects, underground and open pit mining, and deep drilling.

58 GEOSCIENCES↗
DOE OSTI2021

Real-time and Autonomous Water Quality Monitoring System Based on Remotely Operated Vehicle

Existing water quality monitoring systems near hydropower facilities are limited by the lack of mobility of the sensors’ carrier platform. Most systems use a buoy, a mounting fixture attached to a solid structure, or a human worker, which significantly limits the selection of the sampling sites and poses safety risks during data collection and equipment maintenance. To improve on this technology, we developed an autonomous water quality monitoring system that can operate in dangerous water environments near hydropower facilities for water sampling at multiple locations. The goal is to enable safe, timely, and comprehensive water-quality data collection; maximize power generation revenue with improved operational control; and reduce Federal Energy Regulatory Commission and state water quality monitoring costs for compliance. The system incorporates a remotely operated vehicle as the mobile monitoring platform, a dissolved oxygen sensor for monitoring water quality, a tether management system for automatically winding the tether, a solar mobile docking platform for suppling power to the ROV, and a web-based graphical user interface for data post-processing and visualization. In addition, preliminary field research are presented to demonstrate the system capabilities.

Salalila, Aljon L.↗
DOE OSTI2020

Temperature Effects on Interdiffusion of Al and U-Mo under Irradiation

A high-energy Xe ion irradiation experiment was conducted to investigate the temperature dependence of interdiffusion in bilayer Al-UMo samples under irradiation. The amount of interdiffusion achieved at a fixed dose with the increase of temperature showed a clear transition at 175 degrees C (with an estimated error in the range of ± 10 degrees C) from temperature-independent to temperature-dependent behavior. The activation energy derived from the curve of interdiffusion quantity vs. irradiation temperature is 0.77 ± 0.16 eV. This information has been utilized to understand the temperature effect on the interdiffusion process that occurred at the interfaces of U-Mo particles and the Al matrix in U-Mo/Al dispersion fuels, whose magnitude significantly impacts the fuel's performance. Although this temperature effect was deemed important, it cannot be examined directly using in-pile irradiation data, as fuel temperatures cannot be measured in reactor irradiation and are highly correlated with fission rate and thermal conductivity evolution. To connect the knowledge accumulated from ion irradiation with in-pile irradiation data, simulation of a full-sized U-Mo/Al dispersion fuel plate irradiated in the FUTURE test in the BR2 reactor was performed with the Dispersion Analysis Research Tool (DART), a dispersion fuel performance code. DART is equipped with an interaction or interdiffusion layer (IL) growth correlation formulated to describe the temperature dependence of ion mixing results. The agreement between calculated and measured fuel meat constituent volume fractions and swelling data demonstrated that the temperature effect on in-pile Al-UMo interdiffusion is well captured with the correlation. In this case, the fitted activation energy is 0.70 eV. Considering the uncertainties associated with the ion irradiation data, the activation energy obtained from in-pile data fitting is in accord with that from ion irradiation results.

36 MATERIALS SCIENCE↗
DOE OSTI2025

Pooled PPIseq: Screening the SARS-CoV-2 and human interface with a scalable multiplexed protein-protein interaction assay platform

Protein-Protein Interactions (PPIs) are a key interface between virus and host, and these interactions are important to both viral reprogramming of the host and to host restriction of viral infection. In particular, viral-host PPI networks can be used to further our understanding of the molecular mechanisms of tissue specificity, host range, and virulence. At higher scales, viral-host PPI screening could also be used to screen for small-molecule antivirals that interfere with essential viral-host interactions, or to explore how the PPI networks between interacting viral and host genomes co-evolve. Current high-throughput PPI assays have screened entire viral-host PPI networks. However, these studies are time consuming, often require specialized equipment, and are difficult to further scale. Here, we develop methods that make larger-scale viral-host PPI screening more accessible. This approach combines the mDHFR split-tag reporter with the iSeq2 interaction-barcoding system to permit massively-multiplexed PPI quantification by simple pooled engineering of barcoded constructs, integration of these constructs into budding yeast, and fitness measurements by pooled cell competitions and barcode-sequencing. We applied this method to screen for PPIs between SARS-CoV-2 proteins and human proteins, screening in triplicate >180,000 ORF-ORF combinations represented by >1,000,000 barcoded lineages. Our results complement previous screens by identifying 74 putative PPIs, including interactions between ORF7A with the taste receptors TAS2R41 and TAS2R7, and between NSP4 with the transmembrane KDELR2 and KDELR3. We show that this PPI screening method is highly scalable, enabling larger studies aimed at generating a broad understanding of how viral effector proteins converge on cellular targets to effect replication.

60 APPLIED LIFE SCIENCES↗