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NASA NTRS2020

BioSentinel ISS Control Experiment Investigation Summary Form

The main science objective of BioSentinel is to develop biosensors to study the effects of the space radiation environment on biological organisms. Given its similarities to human cells, we decided to use the budding yeast Saccharomyces cerevisiae. Yeast cells share homology to human cells, particularly in the response to DNA damage caused by ionizing radiation. Most importantly, yeast cells can be desiccated and survive in that form for long periods of time. Thus, budding yeast is the ideal model organism for long-term experiments in preparation for future manned missions to deep space. In addition to the ISS mission, our biosensors will fly in a nanosatellite designed for deep space, which is manifested to fly as a secondary payload onboard NASA’s Artemis-1 rocket.

BioSentinel↗

NASA NTRSDate not supplied

BioNutrients-1: Development of an On-Demand Nutrient Production System for Long-Duration Missions

Future long-duration missions beyond low-Earth orbit will require advances in food technologies to address the documented problem of degradation of vitamins and nutrients in supplied foods stored long-term. To begin to address the issue of nutrient degradation, we are developing and flight-testing a platform biomanufacturing technology for in situ production of target nutrients. This technology is being tested over a five-year duration on the International Space Station (ISS). As part of the BioNutrients-1 project we have developed an on-demand system for the production of two carotenoids, β-carotene and zeaxanthin, by genetically engineering distinct strains of Saccharomyces cerevisiae, more commonly known as baker’s yeast. The on-orbit nutrient production packs contain a desiccated yeast strain and edible growth substrate. Once hydrated, the contents of the production packs are intended to grow and produce a desired amount of ready-to-consume nutrients. In this current version the production packs will not be consumed and future missions will require an inactivation of microorganisms before consumption. In addition to the on-orbit hydration of the production packs a series of valuable microorganisms are currently being stored in stasis packs on the ISS including probiotics organisms, bacterial strains used in yogurt production, and organisms with potential use for future biomanufacturing. Analysis of returned ISS stasis packs and ground controls will include multi-omics studies and provide insight into long-term survival of organisms stored in a space environment. Both stasis packs and hydrated production packs will be intermittently returned to Earth for analysis. Preliminary data from long-term storage studies of stasis packs stored on the ISS for 47 days versus their ground control counterparts have shown no significant difference in viability. Currently no production packs have been processed.

Yeast↗

NASA NTRSDate not supplied

Investigating the Molecular Response in Irradiated Yeast in Support of the BioSentinel CubeSat Mission

The BioSentinel CubeSat mission is NASA’s first deep space bioscience mission scheduled to fly as a secondary payload onboard Artemis I. BioSentinel seeks to understand the impacts of deep space radiation on living organisms using the model organism Saccharomyces cerevisiae, or budding yeast. In recent years, the BioSentinel team has conducted ground-based investigations into the effects of ionizing radiation (IR) on budding yeast to better understand spaceflight data. One investigation focuses on the impacts of radiation on budding yeast metabolics using the redox dye alamarBlue, an indicator that will also be used onboard BioSentinel. In this investigation, we have observed metabolic changes in both wild-type and in mutant yeast cells deficient in DNA repair. The focus of our research is to support the ongoing investigation into the molecular mechanisms that explain these observed changes. To do this, we first conducted preliminary research into the metabolic processes of S. cerevisiae to identify any potential vulnerabilities to radiation. We then conducted a statistical kinetic analysis on past experimental metabolic data, in addition to an analysis of data from a multisensor metabolic rig experiment. Ultimately, we identified numerous redox-related changes with increasing doses of radiation in both wild-type and mutant cells. These findings highlight the mitochondrial Electron Transport Chain (ETC) as a potential candidate process to account for these observed changes. Specifically, we are focused on the electron carriers of the ETC, including NADH and FADH2, given their participation in numerous redox mechanisms. We propose future experiments, including NADH assays, an investigation into the glycerol-3-phosphate shuttle, and investigations into other ETC complexes.

BioSentinel↗

NASA NTRSDate not supplied

Improving Data Analyses for a Biological Mission to Deep Interplanetary Space

Onboard the Artemis 1 rocket, NASA plans to launch the first deep space bioscience mission past low Earth orbit (LEO) since 1972, BioSentinel, a 6-Unit (6U) biological CubeSat. BioSentinel’s goals are to assess the effect of deep space ionizing radiation (IR) on DNA and cell damage response, using Saccharomyces cerevisiae, or budding yeast, as a model organism. BioSentinel accomplishes this by measuring Optical Density (OD) in addition to metabolic activity using the redox dye alamarBlue, both of which will be read through light emitting diode (LED) lights of differing wavelengths. With the largest and most sophisticated energy-providing solar panels utilized on a biological CubeSat to date, BioSentinel will be equipped with an IR dosimeter to identify what doses of radiation the yeast is exposed to at any point in time, in addition to a transponder to send such data back to NASA Ames’ Multi Mission Operations Center (MMOC). Biology computational programs and data processing scripts are necessary to analyze this complex data once it is received, including automated subroutines to analyze duplication rate, alamarBlue reduction, and time periods when paired against numerous other variables. Overall, we have implemented several VisualBasic biocomputational programs to analyze such data in an organized and efficient manner so that conclusions can be made rapidly. Furthermore, we showcase why efficient analysis of particular parameters is important to better understand the risks that deep space IR poses to astronauts, especially when considering the upcoming Artemis missions.

Bijan Harandi↗

NASA NTRSDate not supplied

Enabling Single Cell Research in Hollow Hydrogel Microparticles

Studying biology at the single cell level enables novel discoveries through ultra-high throughput identification and isolation of rare cells and interesting subpopulations. Encapsulation of single cells in hollow hydrogel microparticles, or PicoShells, enables easy visualization, manipulation, and assaying of single cells. The PicoShell’s porous hydrogel shell allows diffusion of nutrients and wastes into and out of the hollow liquid core, allowing uninhibited cell growth. The porous shell also allows diffusion of assay reagents, enabling solution changes, and multi-step assays. Finally, single cells and their progeny can be tracked through time, for example to measure single cell growth rates. However, PicoShells are fabricated using microfluidic techniques, which require specialized expertise, equipment, and facilities. This limits PicoShell’s widespread utility for single cell biology. For Saccharomyces cerevisiae and other desiccation-tolerant microbes, desiccating PicoShells containing cells of interest offers a solution, enabling those without microfluidic capabilities to obtain dried, shelf-stable PicoShells containing their cells of interest from a specialized microfluidics facility. Researchers could then rehydrate the PicoShells and perform their single cell assays, bypassing the microfluidic steps. Desiccated PicoShells could also be flown in space, where desiccation is already a common practice for microbes (i.e. BioSentinel and BioNutrients), bringing single cell analysis capabilities to space. In this work, we develop a method to desiccate PicoShells containing S. cerevisiae while maintaining characteristics similar to fresh, non-desiccated PicoShells. We show good microparticle morphology and hydrogel pore size as well as high yeast viability in line with previous studies. We also demonstrate sorting of a fast-growing yeast population using size-based filtration, as S. cerevisiae in PicoShells are able to physically stretch the microparticle as they grow, increasing the PicoShell diameter. These efforts contribute to the feasibility of leveraging microparticle PicoShells for single cell space biology research on Earth and in space.

Simon Ng↗

NASA NTRSDate not supplied

Microbial Factories for Solar System Exploration

As humans venture deeper into space for longer duration missions, resupply missions become far more difficult and costly. To address this, alternative methods of in situ biomanufacturing can be developed to provide a reliable source of required nutrients. BioNutrients is an ongoing project including three separate flight experiments that aims to provide on-demand nutrient production through 5-year shelf stable microbial factories for astronauts on long-duration missions. The BioNutrients-3 (BN-3) flight experiment aims to demonstrate on-orbit food safety, user-friendly production pack design, and multi-compound production capability. This current study seeks to support the multiple compound production element of BN-3 by investigating the production of multiple compounds via multiple pathways in one organism. An experiment has been designed to modify an existing strain of Saccharomyces boulardii designed by the BioNutrients team to efficiently produce β-carotene to also efficiently produce a second nutrient, riboflavin. This experiment entails the random shuffling and recombination of gene elements including promoters and terminators of various strengths to identify optimal combinations for the upregulation of the riboflavin production pathway. Implementation of this experiment would provide insight into the efficiency of the multiple pathways production method and pave the way for additional fine tuning of the riboflavin production pathway and additional nutrient production pathways in S. boulardii.

Biomanufacturing↗

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 NTRSDate not supplied

Remodeling BioSentinel’s Microbial Metabolism Measurement System

BioSentinel, the first biological CubeSat beyond low Earth orbit, will investigate how Saccharomyces cerevisiae responds to ionizing radiation using optical measurements of a colorimetric metabolic indicator dye (alamarBlue). To map optical data to more detailed metabolic measurements, a microbial culture system for ground testing was designed consisting of a 400 mL beaker and a rubber stopper with pass-through holes for its five sensor probes (electrical conductivity, pH, oxidation-reduction potential, dissolved oxygen, optical density). The initial design had several drawbacks. It was only partially reusable due to its one-time sealants: Teflon tape and parafilm. A steady increase in oxygen levels within the sensor’s testingdata indicated the design was not sufficiently airtight. Degradation of the rubber stopper’s elasticity due to sterilization and reinsertion of the probes led to poor robustness and eventual replacement. The vessel’s large headspace didn’t reproduce flight conditions and a large culture volume required more reagents. The system has been redesigned to contain a rigid, biocompatible polycarbonate disc lid that allows for threaded inserts. The culture vessel’s height will be reduced from 110 mm to 65 mm to decrease overhead space and material costs. A more reliable seal will be achieved with a gasket and clamps. The optical probe records absorbance at four wavelengths to characterize cell density and alamarBlue dye ratios. A pneumatically operated plunger draws media through the hollow interior of the probe. Currently, deformation of the silicone on the plunger causes asymmetrical movement and a drift from its initial state. Designs are being developed to limit the plunger’s travel for a more accurate measurement cycle. The remodeled system is under construction and will be tested for leaks and accuracy. The measurements collected will be used to provide additional insight into the optical data returned from the BioSentinel mission.

Astrobiology↗

NASA NTRSDate not supplied

Tracking Metabolic Changes in Microbial Culture using Redox Measurements

During long-term space missions, microbial cultures accumulate the effects of low-dose radiation, microgravity, and other factors; altered growth and metabolic activity may occur before viability effects. This could affect functionality of bioreactors or other bio-enabled mission systems, as well as shed light on human health. Spaceflight microbiology studies beyond the low Earth orbit exposure afforded by the ISS have been limited. Nanosatellites offer an increasingly popular alternative for deep space missions. However, the communications delay requires biofluidic automation of a pre-defined experimental protocol, and the lack of sample return (reliance on sensors in flight) can significantly limit feasible investigations. Previous biological CubeSats (PharmaSat, O/OREOS, EcAMSat) have used alamarBlue, an off-the-shelf formulation of the redox indicator dye resazurin, to track metabolic activity, as will BioSentinel, the upcoming interplanetary microbiology experiment. A series of ground experiments (see abstracts by Liddell, Santa Maria, and A. Kim) were conducted using a microbial culture system outfitted with an electrochemical sensor array (electrical conductivity, pH, oxidation-reduction potential, and dissolved oxygen) with alamarBlue and the same strain of Saccharomyces cerevisiae as BioSentinel. By improving mapping of measured changes in alamarBlue kinetics to physicochemical changes, and ultimately to biological alterations such as shifted metabolic pathways, this work supplements data analyses from past missions and planning for future missions using alamarBlue to characterize space radiation effects. Initial results indicate that alamarBlue acts like a redox buffer; its presence significantly changes redox kinetics in otherwise identical cultures. The initial color change (blue resazurin reduced to red/pink resorufin) appears as a redox plateau. A second plateau, likely corresponding to the second color transition (resorufin to the colorless hydroresorufin), occurs at a lower redox value. The relationship to carbon source exhaustion, dissolved oxygen depletion, cell death, and measured redox potential is complex and still under study.

Tracking↗

NASA NTRSDate not supplied

AMMPER: Agent-based Model for Microbial Populations Exposed to Radiation

Exposure of microbial populations to galactic cosmic radiation (GCR) in the deep-space radiation environment may lead to intracellular damage that compromises the ability of cells to repair and replicate. While simulation programs of deep-space radiation do exist, they focus primarily on single-cell damage, rather than population-wide effects. In this work, a new application and graphical user interface, Agent-Based Model for Microbial Populations Exposed to Radiation (AMMPER) is presented, which simulates the effects of proton-based GCR on Saccharomyces cerevisiae population growth. AMMPER consists of a 50x50x50 µm simulation space, analogous to an aqueous culture medium with non-limiting nutrient and pH buffering, in a microwell plate or microfluidic culture card. This model utilizes Relativistic Ion Tracks (RITRACKS) to create detailed track structures of the radiation traversals. AMMPER then calculates the radiation dose present at each cell, and subsequently determines the damage (chromosomal aberrations, oxidative stress, etc.) and resulting loss of cell viability from both primary and secondary radiative effects. Through implementing cell replication, repair, damage, and death, the effect of radiation exposure on the population growth can be determined. With AMMPER, long-duration effects of the deep space environment on entire populations can be determined and used to assess the feasibility of sustaining life in space.

Amrita Singh↗

NASA NTRSDate not supplied

BioSentinel: To the Moon or Beyond?

BioSentinel, an Artemis-1 secondary spacecraft, will carry a biology experiment into deep space for the first time in 50 years. A 6U CubeSat form factor was utilized for the spacecraft and included technologies newly developed or adapted for operations beyond Earth orbit. This is the maiden deep-space voyage for the radio, propulsion system, electrical power system, and BioSensor payload. The spacecraft carries onboard budding yeast, Saccharomyces cerevisiae, as an analog to human cells to test the biological response to deep space radiation. Flying a secondary payload beyond LEO comes with unique challenges with respect to trajectory uncertainty and mission operations planning. BioSentinel does not carry propulsion for trajectory maneuvers, so plans for Comms and Power need to be developed for all trajectories. The nominal plan is a lunar flyby followed by an insertion into Heliocentric orbit. However, some possible scenarios include lunar eclipses that could severely impact the power budget during that phase of the mission, while others could result in a “Retrograde” hyperbola at swingby resulting in the spacecraft traveling inward toward Earth or even towards a collision with the lunar surface. BioSentinel’s final trajectory will not be known until after launch and deployment so possible scenarios need to be planned for ahead of time. This paper discusses the operational scenarios that were planned for as well as the actual execution of the mission operations including: command pass scheduling with the Deep Space Network, selecting bandwidth limits, medium gain antenna versus low gain antenna usage, and conserving power prior to a lunar eclipse. Note: Artemis-1 is planned to launch in the Spring of 2022

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 and ISS ground control segments.

BioSentinel↗

NASA NTRSDate not supplied

BioSentinel ISS mission: Analysis of ISS Flight Data and Lessons Learned

As we prepare for a future with a human presence on the Moon, Mars, and beyond, the need for countermeasures to protect astronauts against deep space radiation is ever pressing. However, our understanding of how life operates in the space environment, especially past the protection of the Van Allen radiation belts, is critically limited. BioSentinel, a biological CubeSat, aims to further investigate the effects of deep space ionizing radiation, utilizing the budding yeast Saccharomyces cerevisiae to examine the cell’s DNA damage response. The yeast wild type and a rad51Δ mutant strain defective for DNA damage repair will be monitored via the redox dye alamarBlue and a 3-color LED detection system. Selected as a secondary payload on Artemis I, BioSentinel will be the first deep space biological experiment in a half-century and the first biological CubeSat or free-flyer to ever reach a heliocentric orbit. Notably, BioSentinel is the only biological CubeSat to include an ISS control study in addition to a ground control. Comparisons between the ground, ISS, and free-flyer experiments will allow us to isolate the effects of deep space radiation from those of microgravity. The ISS study also provides a unique platform to conduct important technological and biological testing of BioSentinel’s instrumentation in preparation for the deep space mission. In this work, we use a series of data processing tools and scripts to analyze ISS flight data as well as samples exposed to simulated space radiation at Brookhaven National Laboratory, specifically looking at cell growth, metabolic activity, and duplication rates based on optical absorbance and alamarBlue kinetics. These analyses provide a crucial set of controls and a framework for analyzing and interpreting future data sets from the free-flyer, helping us gain further insight into the health risks astronauts will face when exposed to deep space radiation.

Kylie Lauren Lo-Wen Akiyama↗

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 NTRSDate not supplied

BioSentinel SmallSat 2022 Presentation

BioSentinel↗

NASA NTRSDate not supplied

BioSentinel: NASA’s First Deep Space Biological Mission

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 NTRSDate not supplied

Spaceflight Autonomous Multigenerational Microbial Sequencer in Support of Plant-Growth Systems

The CubeSat platform has proven successful in obtaining meaningful life science information when biological payloads are incorporated. Examples include: 1) the first-ever CubeSat with a biological payload, GeneSat-1, which demonstrated decreased growth rates for flight samples of Escherichia coli in low Earth orbit (Parra et al. 2008); 2) PharmaSat, demonstrated that Saccharomyces cerevisiae in the microgravity environment exhibits a significant level of metabolic activity even at high doses of applied antifungal (Ricco et al. 2011); 3) O/OREOS, which used Bacillus subtilis(bacteria) to demonstrate for the first time that microorganisms can be loaded in a dried, dormant form and then rehydrated and grown in orbit months after launch (Nicholson et al. 2011; Ehrenfreund et al. 2014; 4) the SporeSat payload, which investigated Ceratopteris richardii(fern spores) using lab-on-a-chip devices (BioCDs) and minicentrifuges to produce artificial gravitational forces in ground studies (Park et al. 2017), with demonstration of the BioCD and minicentrifuge in space; 5) EcAMSat, the first CubeSat to be directly deployed from the ISS for an experiment assessing antibiotic resistance of E. coli in the microgravity environment (Padgen et al. 2020); 6) BioSentinel, exposed a culture of yeast to galactic cosmic radiation (GCR) and solar particle events while in heliocentric orbit to measure the rate of double-strand-break repair using DNA-repair-deficient mutants. This effort measures the metabolic parameters of yeast in a deep-space environment compared to Earth ambient conditions using a 3-color LED detection system (Ricco et al. 2020; Padgen et al. 2021). We aim to expand this list to include a Spaceflight Autonomous Multigenerational Microbial Sequencer (SAMMS). SAMMS will allow for the genome level understanding of changes in growth and metabolic activity for any organism. While microbes are suitable for early studies in our proposed platform because of their small size, small and relatively less-complicated genomes, fast generation times, and relevance to life support systems; multicellular organisms can similarly be evaluated for their genetic response to the spaceflight environment. The Spaceflight Autonomous Multigenerational Microbial Sequencer (SAMMS) will enable autonomous sequencing of biological samples in plant production units, cislunar orbit and on the lunar surface to examine spaceflight effects (ie. radiation, altered gravity, reduced pressures) on plant and microbial genomes.On this team a Kennedy Space Center (KSC) space crop production and water systems microbiologist/molecular biologist works with a Johnson Space Center (JSC) International Space Station (ISS) microbial sequencing expert and an Ames Research Center (ARC) CubeSat Engineering team to convert an automated Oxford Nanopore librarypreparation and sequencing method to a fluidic CubeSat payload system. The Oxford Nanopore MinION sequencing platform has proven successful in the spaceflight environment onboard the ISS (Stahl-Rommel et al. 2021). Further long-duration spaceflight and exposure to high levels of radiation will cause genotypic effects in biological organisms that may affect their function. Monitoring the adaption of a population to the spaceflight environment and any subsequent beneficial mutations will allow for the harnessing of organisms best suited for use in life support systems. This will ensure that the selected life support-essential microorganisms maintain their intended specified function over generations of culturing in the relevant spaceflight environment without becoming hazardous to crew or spacecraft systems.

Aubrie E Orourke↗