Results for “yeast S. cerevisiae” — Page 2

NASA NTRS2018

BioSentinel: Optimizing Growth Conditions for Improved Yeast Cell Viability After Long-Term Desiccation

NASA's BioSentinel mission is one of thirteen secondary payloads to be deployed on the Space Launch System Exploration Mission-1 (SLS EM-1). The BioSentinel nanosatellite will be sent into a heliocentric orbit beyond Low Earth Orbit (LEO), to study the effects of deep space radiation on the budding yeast, Saccharomyces cerevisiae. Ionizing radiation encountered in deep space can create damaging lesions in DNA, including double strand breaks (DSBs). Budding yeast is suitable as a biological model to study these effects, as it is eukaryotic, and can be desiccated for prolonged periods while retaining viability, thus serving as a robust analog for human cells. On the ground, yeast cells are grown in liquid medium, then loaded into the wells of microfluidic cards and air dried prior to integration into the payload. Once the spacecraft reaches its target heliocentric orbit, a mixture of growth medium and metabolic indicator dye will be pumped into the microwells at specific time points to rehydrate the cells and allow them to grow. A 3-color LED detection system will measure changes in growth and metabolism resulting from ionizing radiation exposure. BioSentinel contains a wild type control strain and a rad51 mutant that is defective for DNA damage repair. In this study, we will determine the optimal amount of time to grow diploid yeast cells in liquid culture before they are desiccated for space flight. After an extended time in stationary phase, they become more tolerant to desiccation due to stress caused by nitrogen starvation. However, excessive exposure can lead to loss of viability and to a heterogeneous cell population due to sporulation. Since viability loss during desiccation poses a risk to mission success, a stress preconditioning process during initial growth may increase long-term cell viability. To determine the growth period that improves desiccation tolerance but allows for retention of uniform radiation sensitivity, we will grow both strains in liquid medium for a varying number of days (4 to 7), desiccate the cells, and then observe changes to cell viability and ionizing radiation sensitivity over time. Supported by the Space Life Sciences Training Program at NASA Ames Research Center.

Cui, Kaixin↗

NASA NTRSDate not supplied

RadBREAD: Radiation Biology Research at an Elevated Altitude through Dosimetry – A student-designed payload

NASA uses extreme environment platforms (ground testing facilities, high-altitude balloons and aircraft, and CubeSats) to provide greater understanding of the conditions and limitations of extra-terrestrial environments. As part of a two-week flight planned for summer 2021, RadBREAD (Radiation Biology Research at an Elevated Altitude through Dosimetry) will fly as a secondary payload consisting of a M-42C (German Aerospace Center, DLR) ionizing radiation dosimeter, UV micro-logger, and multiple desiccated yeast samples. The platform is a novel high-altitude solar-powered aircraft: the Swift Engineering High-Altitude samples. The platform is a novel high-altitude solar-powered aircraft: the Swift Engineering High-Altitude Long-Endurance Unmanned Aircraft System (HALE UAS), which offers significantly longer flight durations than other high-altitude platforms. The yeast Saccharomyces cerevisiae will provide meaningful biological correlation for the sensor readings, due to its resistance to extremely low temperature and pressure when desiccated, ease of genetic manipulation, and homology to human genes. The RadBREAD team comprises the 2020 cohort of NASA’s Space Life Sciences Training Program (SLSTP) research associates as well as NASA scientists, engineers and radiation experts from NASA and the DLR. Yeast survival, metabolic, and transcriptomic changes will be correlated with environmental data collected during long-term exposure to the upper atmosphere. Additionally, the team will evaluate the upper atmospheric environment (radiation, pressure, and temperature) provided by the HALE UAS platform as a Mars surface analog for biological payloads. We hypothesize that exposure to upper atmospheric conditions during the HALE UAS flight will alter the survival, metabolism, and transcriptome of desiccated wild-type S. cerevisiae upon rehydration compared to sensitive and tolerant yeast strains exposed to the same conditions, and between the flight samples compared to asynchronous ground controls.

radiation exposure↗

NASA NTRS2017

Building Better Biosensors for Exploration into Deep-Space, Using Humanized Yeast

1.BioSentinel is 1 of 13 secondary payloads hitching a ride beyond Low Earth Orbit on Exploration Mission 1 (EM-1), set to launch from NASAs Space Launch System in 2019. EM-1 is our first opportunity to investigate the effects of the deep space environment on a eukaryotic biological system, the budding yeast S. cerevisiae. Though separated by a billion years of evolution we share hundreds of genes important for basic cell function, including responses to DNA damage. Thus, yeast is an ideal biosensor for detecting typesextent of damage induced by deep-space radiation.We will fly desiccated cells, then rehydrate to wake them up when the automated payload is ready to initiate the experiment. Rehydration solution contains SC (Synthetic Complete) media and alamarBlue, an indicator for changes in growth and metabolism. Telemetry of LED readings will then allow us to detect how cells respond throughout the mission. The desiccation-rehydration process can be extremely damaging to cells, and can severely diminish our ability to accurately measure and model cellular responses to deep-space radiation. The aim of this study is to develop a better biosensor: yeast strains that are more resistant to desiccation stress. We will over-express known cellular protectants, including hydrophilin Sip18, the protein disaggregase Hsp104, and thioredoxin Trx2, a responder to oxidative stress, then measure cell viability after desiccation to determine which factors improve stress tolerance. Over-expression of SIP18 in wine yeast starter cultures was previously reported to increase viability following desiccation stress by up to 70. Thus, we expect similar improvements in our space-yeast strains. By designing better yeast biosensors we can better prepare for and mitigate the potential dangers of deep-space radiation for future missions.This work is funded by NASAs AES program.

biosensor↗

NASA NTRS2016

Genomic and Phenotypic Characterization of Yeast Biosensor for Deep-space Radiation

The BioSentinel mission was selected to launch as a secondary payload onboard NASA Exploration Mission 1 (EM-1) in 2018. In BioSentinel, the budding yeast Saccharomyces cerevisiae will be used as a biosensor to measure the long-term impact of deep-space radiation to living organisms. In the 4U-payload, desiccated yeast cells from different strains will be stored inside microfluidic cards equipped with 3-color LED optical detection system to monitor cell growth and metabolic activity. At different times throughout the 12-month mission, these cards will be filled with liquid yeast growth media to rehydrate and grow the desiccated cells. The growth and metabolic rates of wild-type and radiation-sensitive strains in deep-space radiation environment will be compared to the rates measured in the ground- and microgravity-control units. These rates will also be correlated with measurements obtained from onboard physical dosimeters. In our preliminary long-term desiccation study, we found that air-drying yeast cells in 10% trehalose is the best method of cell preservation in order to survive the entire 18-month mission duration (6-month pre-launch plus 12-month full-mission periods). However, our study also revealed that desiccated yeast cells have decreasing viability over time when stored in payload-like environment. This suggests that the yeast biosensor will have different population of cells at different time points during the long-term mission. In this study, we are characterizing genomic and phenotypic changes in our yeast biosensor due to long-term storage and desiccation. For each yeast strain that will be part of the biosensor, several clones were reisolated after long-term storage by desiccation. These clones were compared to their respective original isolate in terms of genomic composition, desiccation tolerance and radiation sensitivity. Interestingly, clones from a radiation-sensitive mutant have better desiccation tolerance compared to their original isolate without losing radiation sensitivity. We employed Next-Generation Sequencing technology to better understand this phenotypic variation. Current effort is focusing on the analysis of high-throughput sequencing data to look for genomic changes in these reisolated clones compared to their original isolate.

yeast↗

NASA NTRSDate not supplied

BioSentinel: NASA’s First Deep Space Biological Mission

Since Apollo 17 in 1972, NASA has sent no humans or other biological organisms outside of Earth’s protective magnetosphere. NASA’s current Artemis program plans to put astronauts back on the Moon and eventually land human missions on Mars. One of the major challenges to long-duration crewed travel and habitation in deep space is an in-depth understanding of the biological effects of space radiation, often convoluted by the impact of reduced gravity. Such missions will require significant countermeasures, likely both technological and biomedical, to protect organisms from chronic radiation exposure. Small satellite missions like CubeSats can inform these countermeasures by investigating model organisms in relevant space environments. The BioSentinel mission is comprised of four segments developed at NASA Ames Research Center: a 6U CubeSat (1U = 10-cm cube), an ISS payload launched in December 2021 and two ground units, one for the mission’s CubeSat and one for the ISS payload. The last three segments have been operational since January 2022 and serve as experimental controls. BioSentinel’s 6U CubeSat is planned to launch as a secondary payload on the Artemis-1 rocket. It will be deployed on a lunar fly-by trajectory and into a heliocentric orbit. BioSentinel will be the first interplanetary satellite to study the biological response to space radiation outside Low Earth Orbit (LEO) in almost 50 years. BioSentinel is a complete, autonomous spacecraft capable of conducting experiments in deep space. Its 4U BioSensor payload is a fully automated and adaptable platform that can perform biological measurements with a range of microorganisms in multiple space environments, including the ISS, free flyers, and other platforms like the Lunar Gateway and lander vehicles. Once it reaches its orbit, BioSentinel’s CubeSat will measure the DNA damage response to ambient radiation in a model organism, the budding yeast Saccharomyces cerevisiae, which will be compared to information provided by an onboard radiation sensor and to data obtained in LEO (on ISS) and on Earth. Once in interplanetary space, fluidic cards containing desiccated yeast will be activated by growth medium addition at different time points throughout the mission. Growth and metabolic activity will be tracked continuously via optical measurements. This paper describes BioSentinel’s objectives, science, data management, and preliminary results from the ISS and ISS ground control segments.

BioSentinel↗

NASA NTRSDate not supplied

BioSentinel: NASA’s First Deep Space Biological Mission

Since Apollo 17 in 1972, NASA has sent no humans or other biological organisms outside of Earth’s protective magnetosphere. NASA’s current Artemis program plans to put astronauts back on the Moon and eventually land human missions on Mars. One of the major challenges to long-duration crewed travel and habitation in deep space is an in-depth understanding of the biological effects of space radiation, often convoluted by the impact of reduced gravity. Such missions will require significant countermeasures, likely both technological and biomedical, to protect organisms from chronic radiation exposure. Small satellite missions like CubeSats can inform these countermeasures by investigating model organisms in relevant space environments. The BioSentinel mission is comprised of four segments developed at NASA Ames Research Center: a 6U CubeSat (1U = 10-cm cube), an ISS payload launched in December 2021 and two ground units, one for the mission’s CubeSat and one for the ISS payload. The last three segments have been operational since January 2022 and serve as experimental controls. BioSentinel’s 6U CubeSat is planned to launch as a secondary payload on the Artemis-1 rocket. It will be deployed on a lunar fly-by trajectory and into a heliocentric orbit. BioSentinel will be the first interplanetary satellite to study the biological response to space radiation outside Low Earth Orbit (LEO) in almost 50 years. BioSentinel is a complete, autonomous spacecraft capable of conducting experiments in deep space. Its 4U BioSensor payload is a fully automated and adaptable platform that can perform biological measurements with a range of microorganisms in multiple space environments, including the ISS, free flyers, and other platforms like the Lunar Gateway and lander vehicles. Once it reaches its orbit, BioSentinel’s CubeSat will measure the DNA damage response to ambient radiation in a model organism, the budding yeast Saccharomyces cerevisiae, which will be compared to information provided by an onboard radiation sensor and to data obtained in LEO (on ISS) and on Earth. Once in interplanetary space, fluidic cards containing desiccated yeast will be activated by growth medium addition at different time points throughout the mission. Growth and metabolic activity will be tracked continuously via optical measurements. This paper describes BioSentinel’s objectives, science, data management, and preliminary results from the ISS segment.

BioSentinel↗

NASA NTRS2022

Evolution of Biological Satellites: From Low Earth Orbit to NASA’s BioSentinel Deep Space Mission

NASA has set its sights on human exploration in deep space with the Artemis missions, with an ambitious plan to put astronauts back on the Moon and to eventually land human missions on Mars. Such missions will require significant countermeasures, likely both technological and biomedical, to protect biology from chronic radiation exposure. Small satellites like CubeSats can inform these countermeasures by querying relevant space environments with model organisms over relevant durations. NASA has launched five biological CubeSats into low Earth orbit (LEO) from GeneSat in 2006 to EcAMSat in 2017. Each one of these missions increased our understanding of the effects of spaceflight, while refining technologies and imparting valuable lessons to the next generation of CubeSats. The Artemis I rocket will carry ten CubeSats, each of them with its own objective. One in particular, BioSentinel, will conduct the first study of the biological response to interplanetary space radiation beyond LEO since Apollo 17. Once it reaches its heliocentric orbit – after a short lunar fly-by – BioSentinel will measure the DNA damage response to ambient radiation in a model organism, the budding yeast Saccharomyces cerevisiae, which will be compared to information provided by an onboard radiation sensor and to data obtained in LEO (on ISS) and on Earth. Once in interplanetary space, fluidic cards containing desiccated yeast cells will be activated by growth medium addition at different time points throughout the mission. Growth and metabolic activity will be tracked continuously via optical density. BioSentinel is a complete, autonomous spacecraft capable of conducting experiments in deep space. Its 4U BioSensor payload is a fully automated and adaptable platform that can perform biological measurements with a range of microorganisms in multiple space environments, including the ISS, free flyers, and other platforms like the Lunar Gateway and lander vehicles.

Sergio R Santa Maria↗

NASA NTRS1992

Microgravitational effects on chromosome behavior (7-IML-1)

The effects of the two major space-related conditions, microgravity and radiation, on the maintenance and transmission of genetic information have been partially documented in many organisms. Specifically, microgravity acts at the chromosomal level, primarily on the structure and segregation of chromosomes, in producing major abberations such as deletions, breaks, nondisjunction, and chromosome loss, and to a lesser degree, cosmic radiation appears to affect the genic level, producing point mutations and DNA damage. To distinguish between the effects from microgravity and from radiation, it is necessary to monitor both mitotic and meiotic genetic damage in the same organism. The yeast Saccharomyces cerevisiae is used to monitor at high resolution the frequency of chromosome loss, nondisjunction, intergenic recombination, and gene mutation in mitotic and meiotic cells, to a degree impossible in other organisms. Because the yeast chromosomes are small, sensitive measurements can be made that can be extrapolated to higher organisms and man. The objectives of the research are: (1) to quantitate the effects of microgravity and its synergism with cosmic radiation on chromosomal integrity and transmission during mitosis and meiosis; (2) to discriminate between chromosomal processes sensitive to microgravity and/or radiation during mitosis and meiosis; and (3) to relate these findings to anomalous mitotic mating type switching and ascosporogenesis following meiosis.

Bruschi, Carlo↗

NASA NTRS2014

Biosentinel: Developing a Space Radiation Biosensor

Ionizing radiation presents a major challenge to human exploration and long-term residence in space. The deep-space radiation spectrum includes highly energetic particles that generate double strand breaks (DSBs), deleterious DNA lesions that are usually repaired without errors via homologous recombination (HR), a conserved pathway in all eukaryotes. While progress identifying and characterizing biological radiation effects using Earth-based facilities has been significant, no terrestrial source duplicates the unique space radiation environment.We are developing a biosensor-based nanosatellite to fly aboard NASAs Space Launch System Exploration Mission 1, expected to launch in 2017 and reach a 1AU (astronomic unit) heliocentric orbit. Our biosensor (called BioSentinel) uses the yeast S. cerevisiae to measure DSBs in response to ambient space radiation. The BioSentinel strain contains engineered genetic defects that prevent growth until and unless a radiation-induced DSB near a reporter gene activates the yeasts HR repair mechanisms. Thus, culture growth and metabolic activity directly indicate a successful DSB-and-repair event. In parallel, HR-defective and wild type strains will provide survival data. Desiccated cells will be carried within independent culture microwells, built into 96-well microfluidic cards. Each microwell set will be activated by media addition at different time points over 18 months, and cell growth will be tracked continuously via optical density. One reserve set will be activated only in the occurrence of a solar particle event. Biological measurements will be compared to data provided by onboard physical dosimeters and to Earth-based experiments.BioSentinel will conduct the first study of biological response to space radiation outside Low Earth Orbit in over 40 years. BioSentinel will thus address strategic knowledge gaps related to the biological effects of space radiation and will provide an adaptable platform to perform human-relevant measurements in multiple space environments. We hope that it can therefore be used on the ISS, on and around other planetary bodies as well as other exploration platforms as a self-contained system that will allow us to compare and calibrate different radiation environments.BioSentinels results will be critical for improving interpretation of the effects of space radiation exposure, and for reducing the risk associated with long-term human exploration.

space radiation↗

NASA NTRSDate not supplied

Investigating Biological Responses to Deep Space Radiation for Missions Beyond Low Earth Orbit (LEO) using Yeast

To enable long-term spaceflight missions and establish habitation on the Moon and Mars, we require a comprehensive understanding of the effects of chronic deep space radiation exposure on humans. BioSentinel is NASA’s first biological CubeSat to venture beyond Low Earth Orbit (LEO). It utilizes Saccharomyces cerevisiae (budding yeast) as a model organism to study biological responses to deep space radiation. Yeast share significant genetic homology with humans, including basic cellular metabolism and DNA repair mechanisms. In addition, unlike human cell cultures, yeast can survive the duration and constraints of a deep space mission. BioSentinel measures biological responses using an optical system and alamarBlue oxidation-reduction (redox) dye. Two strains of yeast are studied - a wild-type and a rad51 mutant strain that is deficient in DNA repair. Changes in metabolism and growth are monitored throughout the nominal 6-month mission. Preliminary tests indicate a significant change in the alamarBlue response to low-dose ionizing radiation (IR). Additionally, rad51 cells have shown an IR dose-dependent decrease in glucose uptake and accumulation of oxidized NADH (NAD+). These biomolecules are involved in reactions responsible for basic cell processes, including growth and development, signaling, and respiration. The current study expanded upon previous data by exposing yeast to deep space-relevant radiation. Glucose and NADH/NAD+ assays were conducted on yeast subjected to varying dosages of high-energy Fe-56 and simulated galactic cosmic rays (GCRs). The resulting data was analyzed using Excel and GraphPad Prism. A particular focus was to identify biomolecules resulting from aerobic respiration, which requires the presence of oxygen, or anaerobic processes. As long-term spaceflight missions draw near, it is increasingly important to characterize biological processes affected by the conditions of deep space. Studying biomolecular damage caused by deep space radiation may enable the development of engineering controls or biomedical therapeutics that mitigate health complications for future astronauts.

Kyra Keenan↗

NASA NTRS2017

Biosentinel: Improving Desiccation Tolerance of Yeast Biosensors for Deep-Space Missions

BioSentinel is one of 13 secondary payloads to be deployed on Exploration Mission 1 (EM-1) in 2019. We will use the budding yeast Saccharomyces cerevisiae as a biosensor to determine how deep-space radiation affects living organisms and to potentially quantify radiation levels through radiation damage analysis. Radiation can damage DNA through double strand breaks (DSBs), which can normally be repaired by homologous recombination. Two yeast strains will be air-dried and stored in microfluidic cards within the payload: a wild-type control strain and a radiation sensitive rad51 mutant that is deficient in DSB repairs. Throughout the mission, the microfluidic cards will be rehydrated with growth medium and an indicator dye. Growth rates of each strain will be measured through LED detection of the reduction of the indicator dye, which correlates with DNA repair and the amount of radiation damage accumulated. Results from BioSentinel will be compared to analog experiments on the ISS and on Earth. It is well known that desiccation can damage yeast cells and decrease viability over time. We performed a screen for desiccation-tolerant rad51 strains. We selected 20 re-isolates of rad51 and ran a weekly screen for desiccation-tolerant mutants for five weeks. Our data shows that viability decreases over time, confirming previous research findings. Isolates L2, L5 and L14 indicate desiccation tolerance and are candidates for whole-genome sequencing. More time is needed to determine whether a specific strain is truly desiccation tolerant. Furthermore, we conducted an intracellular trehalose assay to test how intracellular trehalose concentrations affect or protect the mutant strains against desiccation stress. S. cerevisiae cell and reagent concentrations from a previously established intracellular trehalose protocol did not yield significant absorbance measurements, so we tested varying cell and reagent concentrations and determined proper concentrations for successful protocol use.

BioSentinel↗

NASA NTRS2017

Biosentinel: Improving Desiccation Tolerance of Yeast Biosensors for Deep-Space Missions

BioSentinel is one of 13 secondary payloads to be deployed on Exploration Mission 1 (EM-1) in 2019. We will use the budding yeast Saccharomyces cerevisiae as a biosensor to determine how deep-space radiation affects living organisms and to potentially quantify radiation levels through radiation damage analysis. Radiation can damage DNA through double strand breaks (DSBs), which can normally be repaired by homologous recombination. Two yeast strains will be air-dried and stored in microfluidic cards within the payload: a wild-type control strain and a radiation sensitive rad51 mutant that is deficient in DSB repairs. Throughout the mission, the microfluidic cards will be rehydrated with growth medium and an indicator dye. Growth rates of each strain will be measured through LED detection of the reduction of the indicator dye, which correlates with DNA repair and the amount of radiation damage accumulated. Results from BioSentinel will be compared to analog experiments on the ISS and on Earth. It is well known that desiccation can damage yeast cells and decrease viability over time. We performed a screen for desiccation-tolerant rad51 strains. We selected 20 re-isolates of rad51 and ran a weekly screen for desiccation-tolerant mutants for five weeks. Our data shows that viability decreases over time, confirming previous research findings. Isolates L2, L5 and L14 indicate desiccation tolerance and are candidates for whole-genome sequencing. More time is needed to determine whether a specific strain is truly desiccation tolerant. Furthermore, we conducted an intracellular trehalose assay to test how intracellular trehalose concentrations affect or protect the mutant strains against desiccation stress. S. cerevisiae cell and reagent concentrations from a previously established intracellular trehalose protocol did not yield significant absorbance measurements, so we tested varying cell and reagent concentrations and determined proper concentrations for successful protocol use.

BioSentinel↗

NASA NTRS1988

Nonselective enrichment for yeast adenine mutants by flow cytometry

The expression of certain adenine biosynthetic mutations in the yeast Saccharomyces cerevisiae results in a red colony color. This phenomenon has historically provided an ideal genetic marker for the study of mutation, recombination, and aneuploidy in lower eukaryotes by classical genetic analysis. In this paper, it is reported that cells carrying ade1 and/or ade2 mutations exhibit primary fluorescence. Based on this observation, the nonselective enrichment of yeast cultures for viable adenine mutants by using the fluorescence-activated cell sorter has been achieved. The advantages of this approach over conventional genetic analysis of mutation, recombination, and mitotic chromosomal stability include speed and accuracy in acquiring data for large numbers of clones. By using appropriate strains, the cell sorter has been used for the isolation of both forward mutations and chromosomal loss events in S. cerevisiae. The resolving power of this system and its noninvasiveness can easily be extended to more complex organisms, including mammalian cells, in which analogous metabolic mutants are available.

Non-NASA Center↗

NASA NTRS2016

BioSentinel: Biosensors for Deep-Space Radiation Study

The BioSentinel mission will be deployed on NASA's Exploration Mission 1 (EM-1) in 2018. We will use the budding yeast, Saccharomyces cerevisiae, as a biosensor to study the effect of deep-space radiation on living cells. The BioSentinel mission will be the first investigation of a biological response to space radiation outside Low Earth Orbit (LEO) in over 40 years. Radiation can cause damage such as double stand breaks (DSBs) on DNA. The yeast cell was chosen for this mission because it is genetically controllable, shares homology with human cells in its DNA repair pathways, and can be stored in a desiccated state for long durations. Three yeast strains will be stored dry in multiple microfluidic cards: a wild type control strain, a mutant defective strain that cannot repair DSBs, and a biosensor strain that can only grow if it gets DSB-and-repair events occurring near a specific gene. Growth and metabolic activity of each strain will be measured by a 3-color LED optical detection system. Parallel experiments will be done on the International Space Station and on Earth so that we can compare the results to that of deep space. One of our main objectives is to characterize the microfluidic card activation sequence before the mission. To increase the sensitivity of yeast cells as biosensors, desiccated yeast in each card will be resuspended in a rehydration buffer. After several weeks, the rehydration buffer will be exchanged with a growth medium in order to measure yeast growth and metabolic activity. We are currently working on a time-course experiment to better understand the effects of the rehydration buffer on the response to ionizing radiation. We will resuspend the dried yeast in our rehydration medium over a period of time; then each week, we will measure the viability and ionizing radiation sensitivity of different yeast strains taken from this rehydration buffer. The data obtained in this study will be useful in finalizing the card activation sequence for this mission.

Yeast↗

NASA NTRS2017

EcAMSat and BioSentinel: Autonomous Bio Nanosatellites Addressing Strategic Knowledge Gaps for Manned Spaceflight Beyond LEO

Manned missions beyond low Earth orbit (LEO) require that several strategic knowledge gaps about the effects of space travel on the human body be addressed. NASA Ames Research Center has been the leader in developing autonomous bio nanosatellites, including past successful missions for GeneSat, PharmaSat, and OOREOS, that tackled some of these issues. These nanosatellites provide in situ measurements, which deliver insight into the dynamic changes in cell behavior in microgravity. In this talk, two upcoming bio nanosatellites developed at Ames, the E. coli Antimicrobial Satellite (EcAMSat) and BioSentinel, will be discussed. Both satellites contain microfluidic systems that precisely deliver nutrients to the microorganisms stored within wells of fluidic cards. Each well, in turn, has its own 3-color LED and detector system which is used to monitor changes in metabolic activity with alamarBlue, a redox indicator, and the optical density of the cells. EcAMSat investigates the effects of microgravity on bacterial resistance to antimicrobial drugs, vital knowledge for understanding how to maintain the health of astronauts in long-term and beyond LEO spaceflight. The behavior of wild type and mutant uropathic E. coli will be compared in microgravity and with ground data to help understand the molecular mechanisms behind antibiotic resistance and how these phenotypes might change in space. BioSentinel seeks to directly measure the effects of space radiation on budding yeast S. cerevisiae, particularly double strand breaks (DSB). While hitching a ride on the SLS EM-1 mission (Orions first unmanned mission to the moon) in 2018, BioSentinel will be kicked off and enter into a heliocentric orbit, becoming the first study of the effects of radiation on living organisms outside LEO since the Apollo program. The yeast are stored in eighteen independent 16-well microfluidic cards, which will be individually activated over the 12 month mission duration. In addition to the wild type and radiation-sensitive mutant strains, a BioSentinel strain of yeast has been developed, which requires a DSB to reactivate growth, thereby allowing for a direct measurement of DSBs caused by radiation. These two missions demonstrate the utility of using autonomous nanosatellites to address strategic knowledge gaps in the push to once again extend manned spaceflight beyond LEO.

Padgen, Mike↗

NASA NTRSDate not supplied

Precision Fermentation: Relieving the Crabtree Effect in S. cerevisiae through Genetic Engineering

Nutrient degradation in the current NASA pre-packaged food system poses significant challenges to crew health in missions beyond low earth orbit as certain vitamins and nutrients have been shown to degrade during extended storage, and resupply and fresh food items cannot be readily provided. BioNutrients(BN) uses synthetic biology in an effort to supplement the NASA food system by delivering in situ production of nutrients and therapeutics produced by genetically engineered microbes. One yeast S. cerevisiae strain utilized in BN is genetically modified to produce two types of antioxidants: zeaxanthin and β-carotene. Zeaxanthin is a non-provitamin A carotenoid and has been implicated in eye health and vision performance, and β-carotene is a provitamin A carotenoid. Microbial production of high-value nutrients can aid our understanding and address the issue of nutrient degradation and loss in the NASA food system. Although S. cerevisiae is an excellent candidate for a long shelf life, on-demand nutrient production system, its space applications are limited by the Crabtree effect where respiration is shut down in favor of fermentative metabolism when glucose is present in high concentrations, causing the release of unwanted ethanol. Current and likely future life support systems are negatively impacted by ethanol and ethanol release is strictly limited.To improve feasibility of using S. cerevisiae, we explored ways to reduce the Crabtree effect, thereby limiting its ethanol production. Specifically, we used CRISPR-cas9 to genetically modify various genes in this pathway

E Zaroff↗

NASA NTRS2017

EcAMSat and BioSentinel: Autonomous Bio Nanosatellites Addressing Strategic Knowledge Gaps for Manned Spaceflight Beyond LEO

Manned missions beyond low Earth orbit (LEO) require that several strategic knowledge gaps about the effects of space travel on the human body be addressed. NASA Ames Research Center has been the leader in developing autonomous bio nanosatellites, including past successful missions for GeneSat, PharmaSat, and O/OREOS, that tackled some of these issues. These nanosatellites provide in situ measurements, which deliver insight into the dynamic changes in cell behavior in microgravity. In this talk, two upcoming bio nanosatellites developed at Ames, the E. coli Antimicrobial Satellite (EcAMSat) and BioSentinel, will be discussed. Both satellites contain microfluidic systems that precisely deliver nutrients to the microorganisms stored within wells of fluidic cards. Each well, in turn, has its own 3-color LED and detector system which is used to monitor changes in metabolic activity with alamarBlue, a redox indicator, and the optical density of the cells. EcAMSat investigates the effects of microgravity on bacterial resistance to antimicrobial drugs, vital knowledge for understanding how to maintain the health of astronauts in long-term and beyond LEO spaceflight. The behavior of wild type and mutant uropathic E. coli will be compared in microgravity and with ground data to help understand the molecular mechanisms behind antibiotic resistance and how these phenotypes might change in space. BioSentinel seeks to directly measure the effects of space radiation on budding yeast S. cerevisiae, particularly double strand breaks (DSB). While hitching a ride on the SLS EM-1 mission (Orion's first unmanned mission to the moon) in 2018, BioSentinel will be kicked off and enter into a heliocentric orbit, becoming the first study of the effects of radiation on living organisms outside LEO since the Apollo program. The yeast are stored in eighteen independent 16-well microfluidic cards, which will be individually activated over the 12 month mission duration. In addition to the wild type and radiation-sensitive mutant strains, a BioSentinel strain of yeast has been developed, which requires a DSB to reactivate growth, thereby allowing for a direct measurement of DSBs caused by radiation. These two missions demonstrate the utility of using autonomous nanosatellites to address strategic knowledge gaps in the push to once again extend manned spaceflight beyond LEO.

Padgen, Michael R.↗

NASA NTRS2024

Synthetic Biology to Support Human Exploration of Deep Space

The International Space Station (ISS) has enabled a continuous human presence in space since November 2000. The ISS is in low Earth orbit, facilitating regular resupply missions to deliver air, water, food, spare parts, and science experiments. NASA’s Moon to Mars campaign seeks to return humans to the Moon and prepare for crewed missions to Mars. Increased distance from Earth poses logistical challenges to provide all resources needed by humans for deep space missions. NASA Ames Research Center is conducting a series of synthetic biology projects to test the use of microbes for on-demand biosynthesis of human micronutrients. The BioNutrients spaceflight experiments test an implementation concept to produce fermented food products in which microbe growth enhances micronutrient content. On-demand production of carotenoids was engineered into two yeast (Saccharomyces cerevisiae) strains that have been tested in nearly 5 years of storage. One of the BioNutrients strains will be incorporated into the Lunar Explorer Instrument for Space Biology Applications (LEIA) project. LEIA is developing an instrument suite to be delivered to the lunar south pole region by the Commercial Lunar Payload Services (CLPS) program. The LEIA instrument suite will be used to measure multiple yeast strains for growth, metabolic activity, and synthetic biology-enabled production of carotenoids, while taking real time measurements of biologically relevant radiation exposure on the lunar surface. LEIA data will be used to assess the impact of lunar surface radiation and reduced gravity on the production of engineered traits.

Synthetic Biology↗