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DOE OSTI2020

Safeguards by Design Projects (FY2020 Final Report)

This University Engagement project challenged engineering students at universities, that do not have Bachelor degrees in nuclear engineering but do have research reactors and some nuclear engineering coursework, to incorporate Safeguards by Design concepts into their Senior Capstone Design Project. This University Engagement project was part of the U. S. Department of Energy’s (DOE) National Nuclear Security Administration (NNSA), Office of Defense Nuclear Nonproliferation, Office of International Nuclear Safeguards, Next Generation Safeguards Initiative, Human Capital Development: University Engagement Program. This program exposed university students with Mechanical Engineering majors and Nuclear Engineering minors to the concepts of international nuclear safeguards. FY20, three teams at the University of Rhode Island and one team at the University of Texas - Austin participated in researching, designing, building, and testing projects to support international nuclear safeguards measurements or verification. The projects involved engaging in activities at the university’s research reactors. All the projects engaged students with prototyping a design and/or tool for application at the Universities’ reactor. At the end of the course, most of the students expressed the experience was a positive and they learned more about international nuclear safeguards and applying requirements than they had previously encountered. This school year the projects were further complicated by the COVID-19 pandemic. Both universities cancelled in class room classes, and limited direct student/professor interactions. Furthermore, Los Alamos National Laboratory (LANL) greatly restricted travel, therefore making it impossible to visit the students at the end of the semester for the review of their design projects. The final design and review meeting for the projects happened via the internet. Additionally, while the teams planned to build prototypes this did not happen since there was a social distancing ban on students meeting in person.

21 SPECIFIC NUCLEAR REACTORS AND ASSOCIATED PLANTS↗

DOE OSTI2022

Safeguards by Design Projects (Final Report-FY-22)

This University Engagement project challenged engineering students at universities, that do not have Bachelor degree programs in nuclear engineering but do have research reactors and some nuclear engineering coursework, to incorporate Safeguards by Design concepts into their Senior Capstone Design Project. This University Engagement project was part of the U. S. Department of Energy’s (DOE) National Nuclear Security Administration (NNSA), Office of Defense Nuclear Nonproliferation, Office of International Nuclear Safeguards, Next Generation Safeguards Initiative (NGSI), Human Capital Development (HCD): University Engagement Program. This program exposed university students with Mechanical Engineering majors and Nuclear Engineering minors to the concepts of international nuclear safeguards. In FY22, three teams at the University of Rhode Island and two teams at the University of Texas - Austin participated in researching, designing, building, and testing projects to support international nuclear safeguards measurements or verification. The projects involved engaging in activities at the university’s research reactors. All the projects engaged students with prototyping a design and/or tool for application at the Universities’ reactor. At the end of the course, most of the students expressed the experience was positive and they learned more about international nuclear safeguards and applying requirements than they had previously encountered. This school year the projects were further complicated by the COVID-19 pandemic. Both universities had limited in classes on campus, still relying on Zoom classes, and limited direct student/professor interactions. Furthermore, Los Alamos National Laboratory (LANL) greatly restricted travel, therefore making it impossible to visit the students at the end of the semester for the review of their design projects. The final design and review meeting for the projects happened via meetings over the internet. Additionally, the teams did build and test some prototypes but could only do so in a limited capacity.

42 ENGINEERING↗

DOE OSTI2024

Engineering in Cyber Resilience with Cyber-Informed Engineering

Engineers have super powers to provide cybersecurity resilience with deterministic engineering solutions and to protect systems from the most catastrophic consequences that a cyber saboteur could cause. Come to this session to learn how to use engineering risk management skills to harden your engineered systems from cyberattacks. Objective 1 Identify what system functions could be digitally induced to cause undesired high-impact consequences. Objective 2 Analyze how loss or instability of digital controls in a subsystem could lead to high-impact consequences. Objective 3 Analyze how loss or instability in the digital connectivity between systems could lead to high-impact consequences. Objective 4 Identify engineering controls which could build resilience by eliminating digital loss or instability pathways or reduce the impact of digital loss or instability. This presentation will introduce Cyber-Informed Engineering, described below, and walk participants through specific engineering use cases to show how engineers can consider the potential for cyber sabotage in their existing system designs and enact deterministic engineering-based controls which eliminate pathways for attack or mitigate specific consequences. A wide variety of application use cases will be considered so that audience members can align the material with familiar engineering applications. CIE is an engineering approach that integrates cyber resilience into the conception, design, build, and operation of any physical system that has digital connectivity, sensors, monitoring, or control. CIE offers the opportunity to use engineering to eliminate or mitigate avenues for cyber attack—starting from the earliest stage of design and continuing throughout the system’s lifecycle. Today, engineers and industrial control system (ICS) technicians build engineered systems with specific goals for safety, reliability, and functionality. While systems engineering includes considerable safety and failure mode analysis, cybersecurity risks are often not specifically addressed—particularly the risks of intentional cyber compromise, exploitation, and misuse. Cyber-Informed Engineering pairs well with traditional cyber defenses and offers an extra designed-in protection to eliminate the most catastrophic consequences which can be realized by an adversary should traditional cyber defenses fail.

42 ENGINEERING↗

DOE OSTI2026

Sand-Based Thermal Storage for Building Heating Applications: A District Energy Case Study

Buildings account for 40% of global energy consumption and contribute to 30% of global carbon emissions. As energy from renewable sources increases in availability and building designers push for increased electrification, thermal energy storage (TES) systems will play a crucial role in extending the usable time horizon of renewable energy. While water, molten salt, and phase change materials are typically used for building TES heating applications, silica-sand has emerged as an alternative medium for concentrated solar power applications due to its low cost, wide availability, and comparable system efficiency. This paper proposes a new silica-sand particle-based TES system for building heating applications. In this work, a novel steam plant for district heating applications is first designed to utilize silica-sand TES, which can be used for different district energy systems. To demonstrate the silica-sand TES plant performance, the design is modelled in Modelica based on a case study on the University of Colorado Boulder’s campus. The simulation results show that the sand TES plant is more costly to operate than a gas-boiler based plant due to the low cost of natural gas, while the site EUI and carbon intensity can be improved. This novel system shows initial promise as a low-carbon alternative to conventional natural gas steam boilers but will require further modelling and follow-up research to improve its energy efficiency. An eventual rise in natural gas prices, and reduction of electricity prices, could improve the economic viability of this system.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

DOE OSTI2021

Designer membraneless organelles sequester native factors for control of cell behavior

Subcellular compartmentalization of macromolecules increases flux and prevents inhibitory interactions to control biochemical reactions. Inspired by this functionality, we sought to build designer compartments that function as hubs to regulate the flow of information through cellular control systems. Here we report a synthetic membraneless organelle platform to control endogenous cellular activities through sequestration and insulation of native proteins. We engineer and express a disordered protein scaffold to assemble micron size condensates and recruit endogenous clients via genomic tagging with high-affinity dimerization motifs. By relocalizing up to ninety percent of a targeted enzymes to synthetic condensates, we efficiently control cellular behaviors, including proliferation, division, and cytoskeletal organization. Further, we demonstrate multiple strategies for controlled cargo release from condensates to switch cells between functional states. These synthetic organelles offer a powerful and generalizable approach to modularly control cell decision-making in a variety of model systems with broad applications for cellular engineering.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2020

Assembly of Building Blocks by Double-End-Anchored Polymers in the Dilute Regime Mediated by Hydrophobic Interactions at Controlled Distances

Hierarchical assembly of building blocks via competing, orthogonal interactions is a hallmark of many of nature’s composite materials that do not require highly specific ligand-receptor interactions. To mimic this assembly mechanism requires the development of building blocks capable of tunable interactions. In the present work, we explored the interplay between repulsive (steric and electrostatic) and attractive hydrophobic forces. The designed building blocks allow hydrophobic forces to effectively act at controlled, large distances, to create and tune the assembly of membrane-based building blocks under dilute conditions and affect their interactions with cellular membranes via physical cross-bridges. Specifically, we employed double-end-anchored poly(ethylene glycol)s (DEA-PEGs)—hydrophilic PEG tethers with hydrophobic tails on both ends. Using differential-interference-contrast optical microscopy, synchrotron small angle X-ray scattering (SAXS), and cryogenic electron microscopy, we investigated the ability of DEA-PEGs to mediate assembly in the dilute regime on multiple length scales and on practical time scales. The PEG length, anchor hydrophobicity, and molar fraction of DEA-PEG molecules within a membrane strongly affect the assembly properties. Additional tuning of the intermembrane interactions can be achieved by adding repulsive interactions via PEG-lipids (steric) or cationic lipids to the DEA-PEG-mediated attractions. Further, while the optical and electron microscope imaging methods provided qualitative evidence of the ability of DEA-PEGs to assemble liposomes, the SAXS measurements and quantitative line-shape analysis in dilute preparations demonstrated that the ensemble average of loosely organized liposomal assemblies maintains DEA-PEG concentration-dependent tethering on defined nanometer length scales. For cationic liposome–DNA nanoparticles (CL–DNA NPs), aggregation induced by DEA-PEGs decreased internalization of NPs by cells, but tuning the DEA-PEG-induced attractions by adding repulsive steric interactions via PEG-lipids limited aggregation and increased NP uptake. However, confocal microscopy imaging together with colocalization studies with Rab11 and LysoTracker as markers of intracellular pathways showed that modifying CL–DNA NPs with DEA-PEGs alters their interactions with the plasma and endosomal membranes.

- x-ray scattering↗

DOE OSTI2023

Scale-up Testing of Advanced Polaris Membrane in CO2 Capture Technology

This final technical report describes work conducted by Membrane Technology and Research, Inc. (MTR) for the U.S. Department of Energy (DOE), National Energy Technology Lab (NETL) on the scale-up and testing of advanced Polaris™ membrane CO2 capture technology at the Technology Centre Mongstad (TCM) under award number DE-FE0031591. The work was performed from August 1, 2018 through January 31, 2023. The overall goal of this project was to design, build and operate an advanced Polaris membrane CO2 capture system at TCM. MTR was assisted in this project by Trimeric Corporation (Trimeric), an engineering design services company, the Carbon Capture Simulation for Industry Impact (CCSI2), a partnership among national laboratories, industry, and academic institutions, and the Technology Centre Mongstad (TCM), who provided the host site for the slipstream field test. This report details the work conducted to scale-up MTR’s second-generation (Gen-2) Polaris membrane and advanced planar membrane modules to a final form factor optimized for commercial use; validate their performance in an engineering-scale field test at TCM; and to show the potential of the MTR process to meet DOE CO2 capture targets from large source point emitters. Work for this project included membrane optimization and scale-up, advanced planar module design and fabrication, design and fabrication of an engineering-scale field test membrane skid, operation of the field test skid processing Residue Fluid Catalytic Cracker (RFCC) industrial flue gas at TCM, and a detailed techno-economic analysis (TEA) of the MTR membrane post-combustion process for CO2 capture. This project validated recent membrane technology advancements at the engineering-scale, moves the MTR advanced post-combustion capture technology to TRL-6, and mitigates risk in future Large Pilot or Demonstration scale-up activities.

20 FOSSIL-FUELED POWER PLANTS↗

DOE OSTI2020

Highly Integrated WBG Power Module for Next Generation Plug-In Vehicles

Current inverter technologies used in electric vehicles (EVs) are bulky, heavy, and expensive. The performance of the inverter is also limited by the traditional Si-based semiconductor and packaging technology. These constraints are responsible for extra weight and volume in the vehicle that reduces the driving range along with usable space within the vehicle. Moreover, operating temperature restrictions, and boundaries on the inverter location within the vehicle drastically reduces the inverter performance. Thus an improved inverter design is required for next generation electric vehicles. The objective of this project was to research, develop and demonstrate a highly integrated WBG power module targeting the next generation of plugin hybrid electric vehicles (PHEVs). This power module needed to be functionally and mechanically optimized for General Motors (GM) traction inverter architecture. General Motors was the prime contractor, led the project, and oversaw the design, build, and test of the integrated power module. Virginia Polytechnic Institute and State University (VT) performed modeling and simulation tasks as well as power module design support. Monolith Semiconductor, Inc provided SiC dies and consulted on devices for the inverter. Oak Ridge National Laboratory (ORNL) performed power module technology evaluations, design evaluations of interconnect options, and performed material analysis and cross-sectioning of die activities.

33 ADVANCED PROPULSION SYSTEMS↗

DOE OSTI2024

Developing Smart Building Technology Modules to Enhance Workforce Preparedness: A Case for AI-Driven Academic and Professional Education

Smart building technologies are resources that improve building energy efficiency and resilience, reduce carbon emissions, and provide load flexibility to the grid. However, in both academic curricula and building professionals’ continuing education, there is a lack of systematic instruction on methods to integrate multiple energy systems including distributed energy resources (DER), smart building technologies, AI (Artificial Intelligence) tools and key concepts, components, and controls, including “Internet of Things” (IoT) devices. In today’s dynamic workforce, this major gap in smart building technology education prevents stakeholders from being able to attract talent with an understanding and preparation to adopt smart building technologies in building design and operations. A federally funded project included a partnership between Slipstream and Texas A&M University (TAMU) to develop a semester-long smart building curriculum for engineering college students with the ability to adapt the contents for workforce development of professionals in building services. The final product consists of 16 training videos adapted for building professionals and the public. The educational content and training materials cover the benefits of building energy systems, the latest sensor technologies and IoT devices, all with a focus on smart building technologies. The key drivers are on topics related to smart building controls (i.e., energy management information systems), smart building control platforms, cybersecurity, grid-interactive-efficient buildings (GEBs), smart building control methods, and occupant-centric control. Although not explicitly included the technologies nod to the need for AI driven technologies to prepare engineers and industry professionals to be future ready. This paper describes the project approach, provides outlines of the training materials, and identifies lessons learned in creating the content for this course. The authors suggest ways to scale the instruction of smart building concepts to empower the workforce to accelerate the adoption of smart building technologies and AI-based teaching and learning in higher education and building sector.

99 GENERAL AND MISCELLANEOUS↗

DOE OSTI2025

Developing a media formulation to sustain ex vivo chloroplast function

Chloroplasts are critical organelles in plants and algae responsible for accumulating biomass through photosynthetic carbon fixation and cellular maintenance through metabolism in the cell. Chloroplasts are increasingly appreciated for their role in biomanufacturing, as they can produce many useful molecules, and a deeper understanding of chloroplast regulation and function would provide more insight for the biotechnological applications of these organelles. However, traditional genetic approaches to manipulate chloroplasts are slow, and generation of transgenic organisms to study their function can take weeks to months, significantly delaying the pace of research. To develop chloroplasts themselves as a quicker and more defined platform, we isolated chloroplasts from the green algae, Chlamydomonas reinhardtii, and examined their photosynthetic function after extraction. Combined with a metabolic modeling approach using flux-balance analysis, we identified key metabolic reactions essential to chloroplast function and leveraged this information into reagents that can be used in a “chloroplast media” capable of maintaining chloroplast photosynthetic function over time ex vivo compared to buffer alone. We envision this could serve as a model platform to enable more rapid design-build-test-learn cycles to study and improve chloroplast function in combination with genetic modifications and potentially as a starting point for the bottom-up design of a synthetic organelle-containing cell.

Chlamydomonas reinhardtii↗

DOE OSTI2020

Nexus of thermal resilience and energy efficiency in buildings: A case study of a nursing home

Extreme weather events become more frequent and severe due to climate change. Although energy efficiency technologies can influence thermal resilience of buildings, they are traditionally studied separately, and their interconnections are rarely quantified. This study developed a methodology of modeling and analysis to provide insights into the nexus of thermal resilience and energy efficiency of buildings. In this work, we conducted a case study of a real nursing home in Florida, where 12 patients died during Hurricane Irma in 2017 due to HVAC system power loss, to understand and quantify how passive and active energy efficiency measures (EEMs) can improve thermal resilience to reduce heat-exposure risk of patients. Results show that passive measures of opening windows and doors for natural ventilation, as well as miscellaneous load reduction, are very effective in eliminating the extreme dangerous occasions. However, to maintain safe conditions, active measures such as on-site power generators and thermal storage are also needed. The nursing home was further studied by changing its location to two other cities: San Francisco (mild climate) and Chicago (cold winter and hot summer). Results revealed that the EEMs' impacts on thermal resilience vary significantly by climate and building characteristics. The study also estimated the costs of EEMs to help stakeholders prioritize the measures. Passive measures that may not save energy may greatly improve thermal resilience, and thus should be considered in building design or retrofit. Findings from this study indicate energy efficiency technologies should be evaluated not only by their energy savings performance but also by their influence on a building's resilience to extreme weather events.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

DOE OSTI2026

Berkeley eXtensible Environment (BXE) v3

The Berkeley eXtensible Environment (BXE) provides a cloud environment for hardware designers and computer architects to design, build, and simulate their custom architectures on an on-premises FPGA cluster. Utilizing the Chipyard, MoSAIC, and FireSim frameworks, users are provided an environment where they can assemble SoC designs from an existing library of components or import their own source code. Once their designs are ready, they can utilize the FireSim framework provided by BXE to deploy and simulate their designs on the FPGA. Users aren't limited to a single FPGA; they can deploy multiple instances across multiple FPGAs, acting like a rack of servers, or partition their large design across multiple FPGAs, ganging multiple FPGAs into a single simulated system.

Fatollahi-Fard, Farzin↗

DOE OSTI2024

Effect of Heat Treatment on Microstructure and Mechanical Property of 316L Stainless Steel Produced by Laser Powder Bed Fusion

The advanced non-light water reactor designs (Gen IV reactors), including molten salt/ very high temperature/ sodium-cooled and lead-cooled fast reactors, typically operate at higher temperatures and more extreme radiation conditions than light water reactors. An intrinsic part of the deployment and progress of Gen IV reactor designs is selecting the most suitable structural material for a specific application. Additive manufacturing (AM), a fairly new process of making physical, three-dimensional objects from a computer design file, is going to completely change the way of design, build and certify nuclear systems. It offers a range of opportunities to produce complex geometries from existing materials, offers new routes for processing of previously difficult to process materials, allows for design of new high-performance materials, and finally facilitates hybridization of dissimilar materials. This emerging technology has successfully produced cars, wind turbine blade molds and even live cells. It could also open up big opportunities for the nuclear industry to quickly deploy technologies at a fraction of the cost. So far, AM techniques have been preliminarily applied in the field of nuclear reactors, including the classical parts such as the pressure vessel of a small reactor with 508-III steel, the bottom nozzle of a fuel assembly with 304L steel, the fuel cladding with zirconium alloy and the integrated impeller of a pump and the multi-channel valve body with 316L steel [6,7]. The AM applications for operating nuclear reactors started in auxiliary plant components and have slowly migrated to metallic reactors and core components, but many of these are not safety critical components. Although many parts used for nuclear reactors have been fabricated by AM techniques, practical applications in engineering are still a long way off due to the uncertainty factors focused on the processing, material properties, analysis methods and application standards, which feeds the safety and life-cycle of the nuclear reactor. Due to rapid, repeated heating and cooling during production, a high dislocation density was present in the AM material. This microstructure feature is unstable at elevated temperature while high temperature is one of the typical operation environments for nuclear reactors. Thus, it is important to understand the thermal effect on the microstructure of AM material. The objectives of this study are to investigate the effect of heat treatment on the microstructure and mechanical properties of 316L stainless steel produced by laser powder bed fusion additive manufacturing, and to determine an appropriate heat treatment practice that will be applied to the lightweight AM lattice-structured material with the same chemistry. The heat treatment study consisted of annealing the samples at a temperature range of 800 to 1200 oC with a 50 oC increment for different times (1-24 hours), followed by vacuum or air cooling. Microstructural characterization was carried out by Scanning Electron Microscope (SEM). Grain size and crystallographic orientation were investigated by Electron Backscatter Diffraction (EBSD). Vickers hardness tests with a 0.5 kg load were employed to determine the hardness of samples after different heat treatments. After heat treatment, the random crystallographic orientation was preserved, and the volume fraction of high-angle grain boundaries (grain boundary misorientation =15 oC) remained the same. The dislocation density decreased with annealing temperature due to recovery. The fine subgrain structures in the as-printed specimen were quite stable up to 1200 oC. Minimal recrystallization was observed up to 1200 oC. Recrystallization initiated only after 8.5 hours at 1200 oC. The SEM images did not show obvious dependence of microstructure on cooling rate. The hardness of the specimens decreased with increasing annealing temperature as a result of the decrease in dislocation density. It is interesting to note that the AM material showed very similar hardness to the wrought material when annealing at similar temperature, although the microstructures are very different. Annealing at 1050 oC for 1 hour followed by air cooling was selected as the heat treatment procedure for the lattice designed lightweight AM 316L material.

36 MATERIALS SCIENCE↗

DOE OSTI2023

Safeguards by Design Table Top Exercises Final Report (FY 2023)

This University Engagement project challenged engineering students at universities, that do not have bachelor’s degree programs in nuclear engineering but do have research reactors and some nuclear engineering coursework, to develop capacity in Safeguards by Design concepts through the application of Tabletop Exercises. This University Engagement project was part of the U. S. Department of Energy’s (DOE) National Nuclear Security Administration (NNSA), Office of Defense Nuclear Nonproliferation, Office of International Nuclear Safeguards, Next Generation Safeguards Initiative (NGSI), Human Capital Development (HCD): University Engagement Program. This program exposed university students with Mechanical Engineering majors and Nuclear Engineering minors to the concepts of international nuclear safeguards. In FY22, three teams at the University of Rhode Island and two teams at the University of Texas Austin participated in researching, designing, building, and testing projects to support international nuclear safeguards measurements or verification. The projects involved engaging in activities at the university’s research reactors. All the projects engaged students with prototyping a design and/or tool for application at the Universities’ reactor. However, for FY23, the direction of the HCD project had changed to implementing a Tabletop Exercise in Safeguards by Design (SBD). A Tabletop Exercise was not executed during FY23, but relationships with both Universities was maintained and how to integrate the exercise into the curriculum of both programs was determined.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗

DOE OSTI2022

A Scalable Method for Decarbonizing Modular Building Solutions: Preprint

The decarbonization movement emphasizes the shift in focus from energy efficiency to directly reducing global-warming impact. Blokable, LLC, a vertically integrated modular builder with an all-electric portfolio, worked with NREL on a roadmap to decarbonize its high-performance building product at a relative cost advantage by utilizing the learning curves of mass production. Previous decarbonization literature focused on either i) lifecycle assessments, or ii) efficiency measures. These decarbonization exercises were bespoke to individual building projects and did not consider positive feedback loops of builder experience or process repetition. Vertically integrated, prefab builders possess the unique ability to leverage learning and repetition to decarbonize their design-build-operate process. This collaboration between Blokable and NREL resulted in a decarbonization strategy utilizing the company's scaling and production efficiencies, on-site renewable energy and storage, and the projected evolution of building components over time based on trends and emerging legislation. The method developed here encompasses a growing business model, lifecycle carbon assessment, and projected changes in product and grid emissions over time due to existing trends and emerging legislation. This methodology incorporates learning-curve efficiencies gleaned from scaled manufacturing, as well as open-source tools integration for energy and carbon accounting. The output projects and compares cost and carbon savings per modular unit as production increases to 10,000 dwelling units annually over 15 years. The resulting roadmap illustrates a path to roughly 60% carbon savings and beyond-net-zero-energy performance at no incremental cost by 2030. The methodology can be mapped to other integrated or productized builders for methodical decarbonization.

affordable housing↗

DOE OSTI2022

A Scalable Method for Decarbonizing Modular Building Solutions

affordable housing↗

DOE OSTI2022

TEAMER: FOSWEC Mooring Modeling and Analysis, Post Access Report and Data

Floating oscillating surge wave energy converters (FOSWECs) offer several advantages over bottom-hinged oscillating surge wave energy converters, including large wave potential at deep-water sites with fewer permitting and environmental concerns outside territorial waters. As a team, Stevens Institute of Technology, Virginia Tech and Resolute Marine Energy are designing a 100 kW FOSWEC with DOE support (2020-2021) for the PacWave test site "PacWave". The proposed FOSWEC consists of a floating platform, two pivoting flaps, and an innovative power-take-off (PTO). The distance between the two flaps is around half the typical wavelength, resulting in out-of-phase motion and a reduction in motion of the frame and mooring loads. The overall goal of the project is to design, build, deploy and analyze a 1:2 scale (100-kW annual averaged electrical power output) device with reduced levelized cost of energy (LCOE) and peak-to-average power ratio, through the co-design and control of the PTO, WEC, and floating platform. This submission includes a Post Access Report and data for the project of Mooring Modeling and Analysis for Floating Oscillating Surge Wave Energy Converter that Powers Marine Aquaculture of RFTS2 (request for technical support). The data are used to generate all figures in the Post Access Report. Project was a collaboration between Virginia Tech and the National Renewable Energy Lab, with funding from TEAMER.

16 TIDAL AND WAVE POWER↗

DOE OSTI2021

Developing Multi-Gene CRISPRa/I Programs to Accelerate DBTL Cycles in ABF Hosts Engineered for Chemical Production (CRADA 468)

Bacterial metabolism is comprised of large and complex gene networks that can produce valuable chemical products. Sophisticated organism engineering efforts are required to optimize production of high-value compounds from these networks. In principle, synthetic multi-gene transcriptional programs could be constructed to reengineer these networks for efficient industrial chemical production. In practice, however, our incomplete ability to understand and model the underlying networks, combined with our limited ability to predictably control the expression of multiple genes makes achieving this goal difficult. To overcome these challenges, we will combine new CRISPR-Cas multi-gene expression programs with computational modeling, machine learning, and multi-omics data to enhance the efficacy of design-build-test-learn (DBTL) cycles. For industrially promising microorganisms in early stages of development, creating technologies for rapidly engineering complex multi-gene programs could be transformative for accelerating data- and model-driven strain design. New CRISPR-Cas tools allow programmable gene activation (CRISPRa) or repression (CRISPRi) at multiple genes simultaneously, using the catalytically inactive Cas9 protein (dCas9) with guide RNAs that recognize DNA targets through predictable Watson-Crick base pairing. To enable accelerated DBTL cycles, we will combine these technologies with advanced Agile BioFoundry (ABF) capabilities for multi-omics data collection and machine learning. We will demonstrate the immediate applicability of these tools by rapidly improving the production of an industrial aromatic in multiple ABF organisms. We recently identified and optimized new transcriptional activators that can be linked to programmable CRISPR-Cas DNA binding domains to activate gene expression in E. coli. We can now use these CRISPRa tools as generalizable trans-acting regulators for combinatorial multi-gene expression tuning that can be easily transferred to new pathways and networks without additional genome engineering. We anticipate these tools will also transfer to new hosts. We have recently found that CRISPRa systems developed in E. coli can be readily ported to Pseudomonas putida, suggesting that multi-gene CRISPRa/i programs for diverse ABF organisms may be within reach.

59 BASIC BIOLOGICAL SCIENCES↗