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dc.contributor.authorRosenstein, Aaronen
dc.date.accessioned2020-11-06T20:13:20Z
dc.date.available2020-11-06T20:13:20Z
dc.identifier.urihttp://hdl.handle.net/1974/28576
dc.description.abstractThe coming decades will represent a quantum leap in the field of crewed space travel, with planned missions back to the Moon, forward to Mars, and possibly beyond. The substantial biological threats of long-term space exploration are still key barriers to enacting these goals. Microgravity and radiation encountered in space are cellular stressors that could severely compromise the health of future astronauts leaving earth’s orbit. Of specific concern is the impact of these stresses on DNA. The Polymerase Error Rate in Space (PolERIS) experiment was devised to identify whether DNA polymerase enzymes, essential for both replication and repair of the genome, are more prone to errors in microgravity. This research necessitated the development of both novel genetics and engineering-based approaches to conduct experiments in microgravityaboard a parabolic flight plane. It was determined that Klenow fragment (exo+), an E. coli DNA polymerase I derivative with a 3’→5’ exonuclease proofreading domain, was 1.1-fold more prone to both substitution (p = 0.02) and deletion (p = 1.51E-7) errors in microgravity when compared to earth-like gravity. This effect was more pronounced in Klenow fragment (exo-), which does not posess proofreading capability, where substitutions and deletions respectively occurred 2.6-fold (p = 1.98E-11) and 1.5-fold (p = 8.74E-13 ) more frequently in microgravity than earthlike gravity. In addition to ensuring genomic protection in space, the ability for cells to remain viable on long-term space missions may require innovative solutions, such as inducing cellular biostasis for both preservation of cellular isolates and possibly humans in the far future. The Biostasis project of the Defense Advanced Research Projects Agency, USA requires a comprehensive set of assays to quantify various modes of DNA damage potentially incurred during cellular stasis. A research exchange program facilitated early-stage efforts to develop DNA repair pathway-specific cell-based fluorescent biosensors. Thus, together with the PolERIS experiment, this thesis contributes to our current understanding of how polymerases exhibit altered functionality in microgravity, and strategies to detect specific modes of DNA damage with novel cell-based biosensors.en
dc.language.isoengen
dc.relation.ispartofseriesCanadian thesesen
dc.rightsQueen's University's Thesis/Dissertation Non-Exclusive License for Deposit to QSpace and Library and Archives Canadaen
dc.rightsProQuest PhD and Master's Theses International Dissemination Agreementen
dc.rightsIntellectual Property Guidelines at Queen's Universityen
dc.rightsCopying and Preserving Your Thesisen
dc.rightsThis publication is made available by the authority of the copyright owner solely for the purpose of private study and research and may not be copied or reproduced except as permitted by the copyright laws without written authority from the copyright owner.en
dc.subjectMicrogravityen
dc.subjectDNA repairen
dc.subjectDNA polymerasesen
dc.subjectDNA Sequencingen
dc.subjectSpaceen
dc.subjectRadiationen
dc.subjectParabolic flighten
dc.subjectPolymerase fidelityen
dc.subjectSynthetic biologyen
dc.subjectBiosensorsen
dc.subjectBiostasisen
dc.titleDNA Polymerization in Microgravity and the Future of Human Space Travelen
dc.typethesisen
dc.description.degreeM.Sc.en
dc.contributor.supervisorVirginia, Walker
dc.contributor.departmentBiologyen
dc.embargo.termsI want to restrict this thesis because I plan on publishing two of my chapters in separate publications, and as such want to restrict it from public access.en
dc.embargo.liftdate2025-10-30T16:31:12Z
dc.embargo.liftdate2025-11-06T00:34:33Z
dc.degree.grantorQueen's University at Kingstonen


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