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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 415 records · Page 23
All sourcesNASA NTRSDOE OSTI
NASA NTRS2011

MSLICE Sequencing

MSLICE Sequencing is a graphical tool for writing sequences and integrating them into RML files, as well as for producing SCMF files for uplink. When operated in a testbed environment, it also supports uplinking these SCMF files to the testbed via Chill. This software features a free-form textural sequence editor featuring syntax coloring, automatic content assistance (including command and argument completion proposals), complete with types, value ranges, unites, and descriptions from the command dictionary that appear as they are typed. The sequence editor also has a "field mode" that allows tabbing between arguments and displays type/range/units/description for each argument as it is edited. Color-coded error and warning annotations on problematic tokens are included, as well as indications of problems that are not visible in the current scroll range. "Quick Fix" suggestions are made for resolving problems, and all the features afforded by modern source editors are also included such as copy/cut/paste, undo/redo, and a sophisticated find-and-replace system optionally using regular expressions. The software offers a full XML editor for RML files, which features syntax coloring, content assistance and problem annotations as above. There is a form-based, "detail view" that allows structured editing of command arguments and sequence parameters when preferred. The "project view" shows the user s "workspace" as a tree of "resources" (projects, folders, and files) that can subsequently be opened in editors by double-clicking. Files can be added, deleted, dragged-dropped/copied-pasted between folders or projects, and these operations are undoable and redoable. A "problems view" contains a tabular list of all problems in the current workspace. Double-clicking on any row in the table opens an editor for the appropriate sequence, scrolling to the specific line with the problem, and highlighting the problematic characters. From there, one can invoke "quick fix" as described above to resolve the issue. Once resolved, saving the file causes the problem to be removed from the problem view.

Crockett, Thomas M.↗
NASA NTRS2013

Test Waveform Applications for JPL STRS Operating Environment

This software demonstrates use of the JPL Space Telecommunications Radio System (STRS) Operating Environment (OE), tests APIs (application programming interfaces) presented by JPL STRS OE, and allows for basic testing of the underlying hardware platform. This software uses the JPL STRS Operating Environment ["JPL Space Tele com - munications Rad io System Operating Environment,"(NPO-4776) NASA Tech Briefs, commercial edition, Vol. 37, No. 1 (January 2013), p. 47] to interact with the JPL-SDR Software Defined Radio developed for the CoNNeCT (COmmunications, Navigation, and Networking rEconfigurable Testbed) Project as part of the SCaN Testbed installed on the International Space Station (ISS). These are the first applications that are compliant with the new NASA STRS Architecture Standard. Several example waveform applications are provided to demonstrate use of the JPL STRS OE for the JPL-SDR platform used for the CoNNeCT Project. The waveforms provide a simple digitizer and playback capability for the SBand RF slice, and a simple digitizer for the GPS slice [CoNNeCT Global Positioning System RF Module, (NPO-47764) NASA Tech Briefs, commercial edition, Vol. 36, No. 3 (March 2012), p. 36]. These waveforms may be used for hardware test, as well as for on-orbit or laboratory checkout. Additional example waveforms implement SpaceWire and timer modules, which can be used for time transfer and demonstration of communication between the two Xilinx FPGAs in the JPLSDR. The waveforms are also compatible with ground-based use of the JPL STRS OE on radio breadboards and Linux.

Lux, James P.↗
NASA NTRS2013

STS Case Study Development Support

The Shuttle Case Study Collection (SCSC) has been developed using lessons learned documented by NASA engineers, analysts, and contractors. The SCSC provides educators with a new tool to teach real-world engineering processes with the goal of providing unique educational materials that enhance critical thinking, decision-making and problem-solving skills. During this third phase of the project, responsibilities included: the revision of the Hyper Text Markup Language (HTML) source code to ensure all pages follow World Wide Web Consortium (W3C) standards, and the addition and edition of website content, including text, documents, and images. Basic HTML knowledge was required, as was basic knowledge of photo editing software, and training to learn how to use NASA's Content Management System for website design. The outcome of this project was its release to the public.

Rosa de Jesus, Dan A.↗
NASA NTRS2013

Deepak Condenser Model (DeCoM)

Development of the DeCoM comes from the requirement of analyzing the performance of a condenser. A component of a loop heat pipe (LHP), the condenser, is interfaced with the radiator in order to reject heat. DeCoM simulates the condenser, with certain input parameters. Systems Improved Numerical Differencing Analyzer (SINDA), a thermal analysis software, calculates the adjoining component temperatures, based on the DeCoM parameters and interface temperatures to the radiator. Application of DeCoM is (at the time of this reporting) restricted to small-scale analysis, without the need for in-depth LHP component integrations. To efficiently develop a model to simulate the LHP condenser, DeCoM was developed to meet this purpose with least complexity. DeCoM is a single-condenser, single-pass simulator for analyzing its behavior. The analysis is done based on the interactions between condenser fluid, the wall, and the interface between the wall and the radiator. DeCoM is based on conservation of energy, two-phase equations, and flow equations. For two-phase, the Lockhart- Martinelli correlation has been used in order to calculate the convection value between fluid and wall. Software such as SINDA (for thermal analysis analysis) and Thermal Desktop (for modeling) are required. DeCoM also includes the ability to implement a condenser into a thermal model with the capability of understanding the code process and being edited to user-specific needs. DeCoM requires no license, and is an open-source code. Advantages to DeCoM include time dependency, reliability, and the ability for the user to view the code process and edit to their needs.

Patel, Deepak↗
NASA NTRS2013

Making the Implicit Explicit: Towards an Assurance Case for DO-178C

For about two decades, compliance with Software Considerations in Airborne Systems and Equipment Certification (DO-178B) has been the primary means for receiving regulatory approval for using software on commercial airplanes. A new edition of the standard, DO-178C, was published in December 2011, and regulatory bodies have started the process towards recognizing this edition. The stated purpose of DO-178C remains unchanged from its predecessor: providing guidance “for the production of software for airborne systems and equipment that performs its intended function with a level of confidence in safety that complies with airworthiness requirements.” Within the text of the guidance, little or no rationale is given for how a particular objective or collection of objectives contributes to achieving this purpose. Thus the assurance case for the document is implicit. This paper discusses a current effort to make the implicit explicit. In particular, the paper describes the current status of the research seeking to identify the specific arguments contained in, or implied by, the DO-178C guidance that implicitly justify the assumption that the document meets its stated purpose.

Holloway, C. Michael↗
NASA NTRS2014

The Fleet Application for Scheduling and Tracking (FAST) Management Website

The FAST application was designed to replace the paper and pen method of checking out and checking in GSA Vehicles at KSC. By innovating from a paper and pen based checkout system to a fully digital one, not only the resources wasted by printing the checkout forms have been reduced, but it also reduces significantly the time that users and fleet managers need to interact with the system as well as improving the record accuracy for each vehicle. The vehicle information is pulled from a centralized database server in the SPSDL. In an attempt to add a new feature to the FAST application, the author of this report (alongside the FAST developers) has been designing and developing the FAST Management Website. The GSA fleet managers had to rely on the FAST developers in order to add new vehicles, edit vehicles and previous transactions, or for generating vehicles reports. By providing an easy-to-use FAST Management Website portal, the GSA fleet managers are now able to easily move vehicles, edit records, and print reports.

Object-Oriented Programming↗
NASA NTRS2015

2015 Annual Report for the Flight Opportunities Program

Welcome to this third edition of the Flight Opportunities program annual report. In this edition, we continue our story of pathfinding NASA's role in the partnership with the U.S. commercial space and space technology R&D communities to advance national space interests and develop technologies critical to NASA's future missions. 2015 was the year in which a planned change to our payload solicitation strategy saw its first tangible result. As you might remember from our 2014 annual report, in 2015 we set out to facilitate a more direct interaction between flight providers and technology developers by providing fixed funding awards to researchers to directly purchase the flight service(s) that best meet their needs. The selection and award of the first six REDDI-F1 flight grants to non-U.S. government researchers was an important milestone in this regard. From now on, using the REDDI-F1 solicitation appendix, the program will enable non-U.S. government researchers to directly purchase flight services on the emerging suborbital market. The same (or similar) commercial flight services will be available to NASA and other U.S. government agencies (OGA) through commercial contracts that NASA has established through our program. For the latter, our program is available to provide campaign management services, similar to the role we play(ed) for technology payloads remaining in our pool from earlier selections. The full impact of this broader strategic change will likely become more visible in the years ahead as our legacy pool gets depleted and we have implemented a new NASA- and OGA-specific call for proposals. One observation that can already be made after two rounds of REDDI-F1 solicitations is that through this change, the list of commercial flight service providers of interest to non-U.S. government researchers has grown from five in 2014 to nine in 2015. On the industry development front, our Announcement of Collaborative Opportunities (ACO) solicitation was promoted to an STMD-wide solicitation and released in 2015 in combination with the Tipping Point solicitation. A total of 22 awards was announced in November 2015, 12 of which are ACO awards, and six of these are funded by Flight Opportunities. Through these ACO awards, the program funds NASA technical expertise and NASA test facilities to aid industry partners in maturing key space technologies, in our case focusing on small launch vehicle technology development. Flight test activity in 2015 saw a steady 13 campaigns with 31 payload-flights (29 unique payloads). Thirteen new payloads were selected into the program in FY2015, and 14 payloads completed flight testing, bringing the total number of completed technology demonstration payloads to 69. Overall, we are pleased with the evolution and growth of the Flight Opportunities program and look forward to continued success in our partnership with the technology R&D community and the commercial space sector.

Flight Testing↗
NASA NTRS2020

Uncertainties in CERES Top-of-Atmosphere Fluxes Caused by Changes in Accompanying Imager

The Clouds and the Earth’s Radiant Energy System (CERES) project provides observations of Earth’s radiation budget using measurements from CERES instruments on board the Terra, Aqua, Suomi National Polar-orbiting Partnership (S-NPP), and NOAA-20 satellites. The CERES top-of-atmosphere (TOA) fluxes are produced by converting radiance measurements using empirical angular distribution models, which are functions of cloud properties that are retrieved from imagers flying with the CERES instruments. As the objective is to create a long-term climate data record, not only calibration consistency of the six CERES instruments needs to be maintained for the entire time period, it is also important to maintain the consistency of other input data sets used to produce this climate data record. In this paper, we address aspects that could potentially affect the CERES TOA flux data quality. Discontinuities in imager calibration can affect cloud retrieval which can lead to erroneous flux trends. When imposing an artificial 0.6 per decade decreasing trend to cloud optical depth, which is similar to the trend difference between CERES Edition 2 and Edition 4 cloud retrievals, the decadal SW flux trend changed from −0.3 5 ± 0.18 Wm −2 to 0.61 ± 0.18 Wm −2 . This indicates that a 13% change in cloud optical depth results in about 1% change in the SW flux. Furthermore, different CERES instruments provide valid fluxes at different viewing zenith angle ranges, and including fluxes derived at the most oblique angels unique to S-NPP (>66 ∘ ) can lead to differences of 0.8 Wm −2 and 0.3 Wm −2 in global monthly mean instantaneous SW flux and LW flux. To ensure continuity, the viewing zenith angle ranges common to all CERES instruments (<66 ∘ ) are used to produce the long-term Earth’s radiation budget climate data record. The consistency of cloud properties retrieved from different imagers also needs to be maintained to ensure the TOA flux consistency.

Wenying Su↗
NASA NTRS2020

State of the Art Small Spacecraft Technology

When the first edition of NASA’s Small Spacecraft Technology State-of-the-art report was published in 2013, 247 CubeSats and 105 other non-CubeSat small spacecraft under 50 kilograms (kg) had been launched, and these represented less than 2% of launched mass into orbit. By 2019, small spacecraft with mass less than 180 kg made up almost 7% of all mass launched into orbit. Additionally, 63% of spacecraft under 600 kg had mass less than 180 kg and of those 47% were CubeSats (1). Since 2013, flight heritage for small spacecraft, primarily CubeSats, has nearly doubled and with dedicated smallsat launch capabilities readily available and expanding, opportunities to demonstrate new technologies and systems are expected to increase. The 2020 edition of this report captures and distills the wealth of new information available on small spacecraft systems from NASA and other publicly available sources. Overall, this report is a survey of small spacecraft technologies sourced from open literature; it does not endeavor to be an original source, and only considers literature in the public domain to identify and classify devices. Commonly used sources for data include manufacturer datasheets, press releases, conference papers, journal papers, public filings with government agencies, news articles, presentations, and Small Spacecraft Systems Virtual Institute Federated Search. Data not appropriate for public dissemination, such as proprietary, export controlled, or otherwise restricted data, are not considered. As a result, this report includes many dedicated hours of desk research performed by subject matter experts reviewing resources noted above.

Bruce Yost↗
NASA NTRS2013

Biomedical Results of the Space Shuttle Program

The Space Shuttle Program (SSP), which was triumphantly completed by the flight of STS-135 on July 21, 2011, holds a special place in the history of Human Space Exploration. For the first time, a vehicle was built that could perform multiple flights in space and return to Earth. Many important questions regarding the ability of humans to adapt and function in space have been answered during the 30 years since the SSP was launched and many lessons have been learned. The multidisciplinary studies conducted during this program allowed NASA to define the normal responses and potential pathologic changes of the major physiological systems to short-duration space flights. The program significantly contributed to the development of in-flight medical care capabilities and multiple countermeasures. A comprehensive system of preflight, in-flight, and post-flight monitoring of the astronauts was built. It is important to emphasize that the SSP was essential for building the International Space Station (ISS) that significantly expanded the boundaries of space research, allowing for planning of long-term near-terrestrial and extraterrestrial missions. As the program was coming to completion, a decision was made by the NASA Lyndon B. Johnson Space Center Human Research Program senior management to summarize the medico-biological results of the SSP in one comprehensive edition. This was quite an ambitious goal. It required selection of a group of experts in different areas to become the chapters lead authors. These experts were charged with the task of reviewing what was learned in the consequent fields to make the chapters a comprehensive source of information for planning of research, operational activities, and medical care for future missions. We hope that we were able to accomplish this goal and the book will serve as a reliable reference edition for the international space biomedicine community, for new generation of space researchers and Flight Surgeons, as well as for all those interested in space exploration. As space exploration continues, the major question of whether humans are capable of adapting to long-term presence and adequately functioning in space habitats will remain open for a long time. we expect that the comprehensive review of the medico-biological results of the SSP along with the data collected during the missions on the space stations (the Russian space station _Mir_ and ISS) provides a good starting point in seeking the answer to this question.

Diana Risin↗
NASA NTRSDate not supplied

Development of a Consistent GEOsat Cloud Property Dataset for the CERES Climate Data Record

Cloud properties are critical for understanding the Earth’s radiation budget and cloud feedbacks. At NASA Langley Research Center, the Satellite ClOud and Radiative Property retrieval System (SatCORPS) provides real-time and historical analyses of clouds derived from Geostationary satellite (GEOsat) data for weather and climate applications. For the Clouds and the Earth’s Radiant Energy System (CERES) program, the global constellation of GEOsats has been analyzed since 2000 to help characterize and account for the diurnal cycle of clouds and their radiative impacts in the CERES climate data record. Obtaining consistent cloud properties over the GEOsat data record during the CERES era is a major objective but a significant challenge considering the diversity of imaging capabilities deployed during that time. The GEOsat data analysis approach for the current CERES Edition-4 (Ed4) data products was focused on accuracy and consistency with MODIS by employing as much spectral information as possible from each satellite. However, the inconsistent use of spectral information across GEOsats led to marked discontinuities in the spatial and temporal record of cloud properties that had to be accounted for post facto in downstream CERES processing. This paper reports progress in developing a new GEOsat analysis system for the next CERES edition (Ed5) that has potential to improve cross-platform consistency and continuity. In this approach, the spectral channel complement is limited to just 3-channels during daytime, ~0.65 µm (VIS), ~3.9 µm (NIR), and ~10.8 µm (IR), common to nearly all of the satellites in the record. At night, a 2-channel approach is taken with the NIR and IR, and ~6.7 µm bands that includes a machine learning approach for optically thick cloud properties. A tradeoff is the potential for reduced accuracy particularly using data from the more advanced satellites that have more spectral channels (e.g. SEVIRI, AHI and ABI) that are known to help improve thin cirrus detection, cloud-aerosol discrimination and estimates in other difficult conditions that challenge cloud remote sensing. The new continuity approach is applied to one month of global GEOSat data for each year of the CERES record since 2000 and compared with the Ed4 GEO and MODIS cloud property time series in order to evaluate the level of improved consistency in the GEOsat record and to assess the accuracy impacts. Cloud fraction will also be assessed with CALIPSO data. Outstanding issues and challenges will be discussed. The results are expected to guide future work needed to develop a more robust GEOsat cloud data record for CERES.

CERES CDR↗
NASA NTRSDate not supplied

Earth Radiation Budget Climate Record Composed of Multiple Satellite Observations

Effects of cloud diurnal cycle on top-of-atmosphere (TOA) and surface regional monthly mean irradiances, climatological mean, and anomalies are analyzed using CERES derived TOA irradiances and surface irradiances computed with MODIS derived cloud properties. Cloud properties derived from Terra and Aqua MODIS are sufficient to capture cloud diurnal cycle to compute regional monthly mean surface irradiances. While missing cloud diurnal cycle leads to a biased TOA and surface regional irradiances for regions with a strong cloud diurnal cycle, monthly regional TOA and surface anomalies derived from one sun-synchronous orbit agrees well with those derived from two sun-synchronous orbits. Based on these results, the algorithm to produce Edition 4.2 CERES EBAF product is developed. Regional TOA and surface climatological means derived from one sun-synchronous orbit are adjusted to match corresponding climatological means derived from Terra+Aqua observations. This climatological adjustment approach is used to merge the Terra only period to the Terra+Aqua period and to extend the Terra and Aqua record by merging NOAA20 observations. Two additional differences of Edition 4.2 EBAF algorithm to compute surface irradiances compared to the earlier version are: 1) no geostationary satellite derived cloud properties are used and 2) temperature and humidity from MERRA-2 instead of GEOIS-5.4.1 are used. Once surface monthly regional mean irradiances are compared with surface observations, the agreement is equivalent to the agreement with the earlier version. However, because surface irradiances are not affected by geostationary satellite artifacts, regional surface irradiance anomaly time series is significantly improved, especially for longwave irradiances.

Seiji Kato↗
NASA NTRSDate not supplied

Progress Towards a Common CERES Cloud Mask Algorithm for MODIS and VIIRS: Evaluation using CALIOP Data

For over two decades The Clouds and the Earth’s Radiant Energy System (CERES) project has endeavored to produce a long-term global climate data record for detecting changes in the Earth’s radiation budget and to improve understanding of how clouds contribute to those changes. CERES incorporates cloud information derived from passive narrowband satellite imaging radiometers, and over the course of the project different instruments have contributed to this effort, namely the Moderate Resolution Imaging Spectroradiometer (MODIS) and the Visible Infrared Imaging Radiometer Suite (VIIRS).Cloud properties have been derived from MODIS on the Aqua satellite for CERES since 2002,and VIIRS on the NOAA-20 satellite launched in 2017 will continue the cloud record once Aqua-MODIS reaches the end of its operational lifetime. However, MODIS and VIIRS have different spectral capabilities and characteristics which complicates our ability to seamlessly transition the record from MODIS to VIIRS. This study evaluates some of the recent algorithm modifications incorporated by the CERES Cloud Working Group towards developing a unified cloud mask algorithm for MODIS and VIIRS that utilizes a set of spectral bands common to both instruments. Cloud-Aerosol Lidar with Orthogonal Polarization (CALIOP) data products are used to evaluate MODIS and VIIRS regional cloud fraction estimates and identify areas where improvements can be made. The cloud mask algorithm relies on computed estimates of cloud-free top-of-atmosphere radiances to differentiate cloudy and cloud-free imager pixels. The unified cloud mask algorithm currently under development uses a reduced number of spectral bands common to both MODIS and VIIRS and so relies on the accuracy of these simulated cloud-free radiances more heavily than predecessor algorithms, i.e., the CERES Edition 4 cloud mask, which use as much spectral information as available on each imager with less consideration for cross-platform consistency. Our validation strategy using CALIOP data is also described. The CALIOP observations are critical for assessing cloud detection accuracies and for independently confirming cloud-free conditions which enables a more robust evaluation of the simulated cloud-free radiances for biases due to factors such as view angle and the spectral dependence of water vapor absorption. These effects are known to differ for the two satellite instruments. Comparisons of MODIS and VIIRS cloud fractions are presented in context with estimates from CALIOP and from the CERES Edition 4 cloud mask algorithm to gauge current progress in developing accurate and consistent MODIS and VIIRS cloud properties for CERES.

CERES↗
NASA NTRS2009

Introduction To Space Shuttle Rendezvous Guidance, Navigation, and Control

The Rendezvous Intro continues to be used for training and reference purposes almost 17 years after it was first published. While this document does not address Orion rendezvous, some Orion personnel are using it as a reference for rendezvous GNC and trajectory concepts. This edition contains corrections that have been identified since the 3rd edition was published in 2000. In addition, new material has been added that was developed by the author in the 1990s and early part of the first decade of the 21st century. While some of this material is perhaps too detailed for an introduction, it is of value and the author wanted to make it available to JSC personnel. The sections on Lambert targeting guidance have been significantly expanded.

John L Goodman↗
DOE OSTI2024

High incidence and geographic distribution of cleft palate in Finland are associated with the IRF6 gene

In Finland, the frequency of isolated cleft palate (CP) is higher than that of isolated cleft lip with or without cleft palate (CL/P). This trend contrasts to that in other European countries but its genetic underpinnings are unknown. We conducted a genome-wide association study in the Finnish population and identified rs570516915, a single nucleotide polymorphism highly enriched in Finns, as strongly associated with CP (P = 5.25 × 10 -34 , OR = 8.65, 95% CI 6.11-12.25), but not with CL/P (P = 7.2 × 10 -5 ), with genome-wide significance. The risk allele frequency of rs570516915 parallels the regional variation of CP prevalence in Finland, and the association was replicated in independent cohorts of CP cases from Finland (P = 8.82 × 10 -28 ) and Estonia (P = 1.25 × 10 -5 ). The risk allele of rs570516915 alters a conserved binding site for the transcription factor IRF6 within an enhancer (MCS-9.7) upstream of the IRF6 gene and diminishes the enhancer activity. Oral epithelial cells derived from CRISPR-Cas9 edited induced pluripotent stem cells demonstrate that the CP-associated allele of rs570516915 concomitantly decreases the binding of IRF6 and the expression level of IRF6, suggesting impaired IRF6 autoregulation as a molecular mechanism underlying the risk for CP.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2025

Data for "Viral Delivery of Recombinases Activates Heritable Genetic Switches in Plants"

Viral vectors provide an increasingly versatile platform for transformation-free reagent delivery to plants. RNA viral vectors can be used to induce gene silencing, overexpress proteins, or introduce gene editing reagents; however, they are often constrained by carrying capacity or restricted tropism in germline cells. Site-specific recombinases that catalyze precise genetic rearrangements are powerful tools for genome engineering that vary in size and, potentially, efficacy in plants. In this work, we show that viral vectors based on tobacco rattle virus (TRV) deliver and stably express four recombinases ranging in size from ∼0.6 to ∼1.5 kb and achieve simultaneous marker removal and reporter activation through targeted excision in transgenic Nicotiana benthamiana lines. TRV vectors with Cre, FLP, CinH, and Integrase13 efficiently mediated recombination in infected somatic tissue and led to heritable modifications at high frequency. An excision-activated Ruby reporter enabled simple and high-resolution tracing of infected cell lineages without the need for molecular genotyping. Together, our experiments broaden the scope of viral recombinase delivery and offer insights into infection dynamics that may be useful in developing future viral vectors.

gene editing↗
DOE OSTI2026

Data for Development, Optimization, and Application of an Episomal Plasmid System for Rhodotorula toruloides

Rhodotorula toruloides is an emerging oleaginous yeast with strong potential as a microbial cell factory for the production of acetyl-CoA-derived bioproducts. However, engineering of this organism has been limited by the absence of a functional episomal plasmid system, a foundational genetic tool for rapid gene expression, pathway testing, and CRISPR-based genome engineering. Here, we report the first episomal plasmid system for R. toruloides . Through systematic screening of candidate autonomously replicating sequences (ARSs) from diverse sources, we identified multiple functional ARS elements and selected C63F4, a fragment derived from Contig 63 of R. toruloides CBS14, because of its stable performance. The resulting pC63F4 plasmid was maintained episomally, supported GFP reporter expression, exhibited a copy number of 2.39 ± 0.13, and showed good stability during long term cultivation. To overcome poor transformation efficiency, we developed a Cre-loxP-mediated in vivo re-circularization strategy that enabled reliable delivery of the episomal plasmid. Using this improved system, we demonstrated functional episomal expression of metabolic engineering genes and multi-gene pathways for the production of triacetic acid lactone, fatty alcohols, and limonene. Finally, we leveraged this platform to establish a redesigned CRISPR system that enables seamless genome editing in R. toruloides for the first time, while also simplifying marker recycling. Together, this work establishes a long-needed episomal plasmid platform and associated CRISPR toolkit that will accelerate metabolic engineering, synthetic biology, and fundamental studies in R. toruloides .

Gene Editing↗
NASA NTRS1999

Proteolytic dissection of Zab, the Z-DNA-binding domain of human ADAR1

Zalpha is a peptide motif that binds to Z-DNA with high affinity. This motif binds to alternating dC-dG sequences stabilized in the Z-conformation by means of bromination or supercoiling, but not to B-DNA. Zalpha is part of the N-terminal region of double-stranded RNA adenosine deaminase (ADAR1), a candidate enzyme for nuclear pre-mRNA editing in mammals. Zalpha is conserved in ADAR1 from many species; in each case, there is a second similar motif, Zbeta, separated from Zalpha by a more divergent linker. To investigate the structure-function relationship of Zalpha, its domain structure was studied by limited proteolysis. Proteolytic profiles indicated that Zalpha is part of a domain, Zab, of 229 amino acids (residues 133-361 in human ADAR1). This domain contains both Zalpha and Zbeta as well as a tandem repeat of a 49-amino acid linker module. Prolonged proteolysis revealed a minimal core domain of 77 amino acids (positions 133-209), containing only Zalpha, which is sufficient to bind left-handed Z-DNA; however, the substrate binding is strikingly different from that of Zab. The second motif, Zbeta, retains its structural integrity only in the context of Zab and does not bind Z-DNA as a separate entity. These results suggest that Zalpha and Zbeta act as a single bipartite domain. In the presence of substrate DNA, Zab becomes more resistant to proteases, suggesting that it adopts a more rigid structure when bound to its substrate, possibly with conformational changes in parts of the protein.

Non-NASA Center↗