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Project Title:  Multi-Generational Genome-Wide Yeast Fitness Profiling Beyond and Below Earth's van Allen Belts Reduce
Images: icon  Fiscal Year: FY 2020 
Division: Space Biology 
Research Discipline/Element:
Cell & Molecular Biology 
Start Date: 05/01/2019  
End Date: 04/30/2022  
Task Last Updated: 02/28/2020 
Download report in PDF pdf
Principal Investigator/Affiliation:   Zea, Luis  Ph.D. / University of Colorado, Boulder 
Address:  Aerospace Engineering Sciences 
429 Ucb, ECAE 1B02 
Boulder , CO 80309-0429 
Email: Luis.Zea@Colorado.edu 
Phone: 407-242-2885  
Congressional District:
Web:  
Organization Type: UNIVERSITY 
Organization Name: University of Colorado, Boulder 
Joint Agency:  
Comments:  
Co-Investigator(s)
Affiliation: 
Stodieck, Louis  Ph.D. University of Colorado, Boulder 
Nislow, Corey  Ph.D. University of British Columbia, Canada 
Project Information: Grant/Contract No. 80NSSC19K0708 
Responsible Center: NASA KSC 
Grant Monitor: Freeland, Denise  
Center Contact: 321-867-5878 
Denise.E.Freeland@nasa.gov 
Solicitation: 2018 Space Biology (ROSBio) NNH18ZTT001N-Artemis1 (EM1). App A: Orion (Artemis-1) (formerly Exploration Mission-1) 
Grant/Contract No.: 80NSSC19K0708 
Project Type: FLIGHT 
Flight Program:  
No. of Post Docs:
No. of PhD Candidates:
No. of Master's Candidates:
No. of Bachelor's Candidates:
No. of PhD Degrees:
No. of Master's Degrees:
No. of Bachelor's Degrees:
Space Biology Element: (1) Cell & Molecular Biology
Space Biology Cross-Element Discipline: None
Space Biology Special Category: None
Task Description: As human space exploration expands beyond lower Earth orbit, it is necessary to characterize the effects of space radiation, microgravity, and the combination thereof on cells. Because it is complicated to have large sample numbers when studying the effects of different factors on humans, scientists commonly use model organisms that share some of the key aspects being studied. In this case, we will use yeast, as around 70% of its essential genes have a significant human homolog. More specifically, this project will use a molecularly barcoded yeast genome-wide knockdown collection that will enable the systematic interrogation of the effect of microgravity, space radiation, and a combination thereof in each gene. Each strain in the collection has a single gene deleted and a representative molecular barcode that enables quantifying the fitness of each mutant under the test conditions, by measuring the relative abundance at different points in time. To differentiate the effects of microgravity and space radiation on each strain, an experimental set will be flown beyond the van Allen belts on Orion’s Exploration Mission 1 (EM-1) (considered in microgravity and irradiated by space radiation) and equivalent sets will be cultured asynchronously on board the International Space Station (ISS) (considered in microgravity but mostly – although not completely – protected of space radiation by the van Allen belts) in our smart incubator (Space Automated Bioproduct Lab (SABL)) and on Earth (also in a ground SABL). Each of the ISS and Earth experiments will include two sets: one where the temperature profile experienced during the EM-1 flight is replicated, and a second cultured at a constant temperature to determine the potential role of temperature variation on the results from EM-1.

The first aim of this project is to identify the metabolic and genomic pathways in yeast affected by microgravity, space radiation, and a combination of both. The second one is to differentiate between gravity and radiation exposure on single-gene deletion mutants’ ability to thrive in the spaceflight environment. We hypothesize that mutants lacking genes associated with DNA repair, recombination, and replication will have lower survivability rates beyond the van Allen belts than their below van Allen belts- or Earth-controls

The experiment is designed to have a controlled start after Orion is past the van Allen belts, grow ~21 generations of the deletion series, and fix or preserve samples for post-flight analyses. Should the automated controlled approach be considered inappropriate for implementation on EM-1, we have a passive approach that is based on dotting each mutant individually on agar. We have performed both approaches in space in the past.

This project will address three Space Biology Program Science Elements, three Objectives, three Guiding Questions, and four Decadal Survey’s highest priority Recommendations by preserving nucleic acids of different generations of the yeast deletion series cultures grown in space, beyond as well as below the van Allen belts (and uploading the genomic and transcriptomic data to GeneLab).

Research Impact/Earth Benefits: This project integrates data on the molecular and cellular mechanism of radiation damage, which can serve to improve prediction of risk of cancer as a function of radiation dosage and to evaluate the effectiveness of potential countermeasures.

Task Progress & Bibliography Information FY2020 
Task Progress: 1. The design of this project’s hardware, Peristaltic Laboratory for Automated Science with Multigenerations (PLASM), matured enough to hold a Critical Design Review (CDR). After that, all of the constituent parts of PLASM have been manufactured and ordered, and are now available for integration and testing.

2. Two irradiation campaigns have taken place to provide data on what may we expect from the Artemis 1 mission, further verifying the validity of our experimental design. Additionally, a 69-day long Science Verification Test (SVT) is currently ongoing and already showing effective yeast growth in flight-like hardware.

Bibliography Type: Description: (Last Updated: 03/05/2020)  Show Cumulative Bibliography Listing
 
Abstracts for Journals and Proceedings Breaux S, Fuchs F, Cortesao M, Siems K, Stodieck L, Niederwieser T, Zea L, Carr CE, Moeller R, Nislow C. "Irradiation Ground Control for a Genome-Wide Yeast Fitness Profiling Experiment On Board Orion’s Artemis 1 Mission." 35th Annual Meeting of the American Society for Gravitational and Space Research, Denver, CO, November 20-23, 2019.

Abstracts. 35th Annual Meeting of the American Society for Gravitational and Space Research, Denver, CO, November 20-23, 2019. , Nov-2019

Papers from Meeting Proceedings Zea L, Niederwieser T, Stodieck L, Carr C, Moeller R, Nislow C. "Experiment Design for a Genome-Wide Yeast Fitness Profiling Experiment On Board Orion’s Artemis 1 Mission." 70th International Astronautical Congress (IAC), Washington, DC, October 21-25, 2019.

70th International Astronautical Congress (IAC), Washington, DC, October 21-25, 2019. Paper IAC-19-A2.7.9x51501. , Oct-2019

Project Title:  Multi-Generational Genome-Wide Yeast Fitness Profiling Beyond and Below Earth's van Allen Belts Reduce
Images: icon  Fiscal Year: FY 2019 
Division: Space Biology 
Research Discipline/Element:
Cell & Molecular Biology 
Start Date: 05/01/2019  
End Date: 04/30/2022  
Task Last Updated: 05/13/2019 
Download report in PDF pdf
Principal Investigator/Affiliation:   Zea, Luis  Ph.D. / University of Colorado, Boulder 
Address:  Aerospace Engineering Sciences 
429 Ucb, ECAE 1B02 
Boulder , CO 80309-0429 
Email: Luis.Zea@Colorado.edu 
Phone: 407-242-2885  
Congressional District:
Web:  
Organization Type: UNIVERSITY 
Organization Name: University of Colorado, Boulder 
Joint Agency:  
Comments:  
Co-Investigator(s)
Affiliation: 
Stodieck, Louis  Ph.D. University of Colorado, Boulder 
Nislow, Corey  Ph.D. University of British Columbia, Canada 
Project Information: Grant/Contract No. 80NSSC19K0708 
Responsible Center: NASA KSC 
Grant Monitor: Freeland, Denise  
Center Contact: 321-867-5878 
Denise.E.Freeland@nasa.gov 
Solicitation: 2018 Space Biology (ROSBio) NNH18ZTT001N-Artemis1 (EM1). App A: Orion (Artemis-1) (formerly Exploration Mission-1) 
Grant/Contract No.: 80NSSC19K0708 
Project Type: FLIGHT 
Flight Program:  
No. of Post Docs:  
No. of PhD Candidates:  
No. of Master's Candidates:  
No. of Bachelor's Candidates:  
No. of PhD Degrees:  
No. of Master's Degrees:  
No. of Bachelor's Degrees:  
Space Biology Element: (1) Cell & Molecular Biology
Space Biology Cross-Element Discipline: None
Space Biology Special Category: None
Task Description: As human space exploration expands beyond lower Earth orbit, it is necessary to characterize the effects of space radiation, microgravity, and the combination thereof on cells. Because it is complicated to have large sample numbers when studying the effects of different factors on humans, scientists commonly use model organisms that share some of the key aspects being studied. In this case, we will use yeast, as around 70% of its essential genes have a significant human homolog. More specifically, this project will use a molecularly barcoded yeast genome-wide knockdown collection that will enable the systematic interrogation of the effect of microgravity, space radiation, and a combination thereof in each gene. Each strain in the collection has a single gene deleted and a representative molecular barcode that enables quantifying the fitness of each mutant under the test conditions, by measuring the relative abundance at different points in time. To differentiate the effects of microgravity and space radiation on each strain, an experimental set will be flown beyond the van Allen belts on Orion’s Exploration Mission 1 (EM-1) (considered in microgravity and irradiated by space radiation) and equivalent sets will be cultured asynchronously on board the International Space Station (ISS) (considered in microgravity but mostly – although not completely – protected of space radiation by the van Allen belts) in our smart incubator (Space Automated Bioproduct Lab (SABL)) and on Earth (also in a ground SABL). Each of the ISS and Earth experiments will include two sets: one where the temperature profile experienced during the EM-1 flight is replicated, and a second cultured at a constant temperature to determine the potential role of temperature variation on the results from EM-1.

The first aim of this project is to identify the metabolic and genomic pathways in yeast affected by microgravity, space radiation, and a combination of both. The second one is to differentiate between gravity and radiation exposure on single-gene deletion mutants’ ability to thrive in the spaceflight environment. We hypothesize that mutants lacking genes associated with DNA repair, recombination, and replication will have lower survivability rates beyond the van Allen belts than their below van Allen belts- or Earth-controls

The experiment is designed to have a controlled start after Orion is past the van Allen belts, grow ~21 generations of the deletion series, and fix or preserve samples for post-flight analyses. Should the automated controlled approach be considered inappropriate for implementation on EM-1, we have a passive approach that is based on dotting each mutant individually on agar. We have performed both approaches in space in the past.

This project will address three Space Biology Program Science Elements, three Objectives, three Guiding Questions, and four Decadal Survey’s highest priority Recommendations by preserving nucleic acids of different generations of the yeast deletion series cultures grown in space, beyond as well as below the van Allen belts (and uploading the genomic and transcriptomic data to GeneLab).

Research Impact/Earth Benefits:

Task Progress & Bibliography Information FY2019 
Task Progress: New project for FY2019.

Bibliography Type: Description: (Last Updated: 03/05/2020)  Show Cumulative Bibliography Listing
 
 None in FY 2019