![]() The CRISPR/Cas9 gene editing system offers a model for the safe and targeted generation of double-strand breaks in eukaryotes. However, our understanding of this problem has been limited by technical and safety concerns, which have prevented integral study of the DNA repair process in space. Previous work suggests that space conditions may impact the choice of DNA repair pathway, potentially compounding the risks of increased radiation exposure during space travel. Double-strand breaks are a type of DNA damage that can be repaired by two major cellular pathways: non-homologous end joining, during which insertions or deletions may be added at the break site, and homologous recombination, in which the DNA sequence often remains unchanged. Ground W1 and W3 had lower coverage mapped to wild type sequence due to the observed deletions ( S1 Fig).Īs we explore beyond Earth, astronauts may be at risk for harmful DNA damage caused by ionizing radiation. Flight W2 observed similar mapping as red colonies with 76% reads mapping to the repair template sequence and 33% of reads mapping to the wild type sequence. With the exception of Flight W2 and Ground W1 and W3, >98% of reads map to the wild type sequence from both flight and ground. In contrast, data from red colony ground controls R1-R4 show that 97.9%, 90.8%, 96%, and 94% of the reads map to the repair template sequence and 10.4%, 16.2%, 11.7%, and 13% reads mapped to wild type sequence. ![]() 63.2%, 58.8%, 72.6%, and 75.1% of the reads from flight samples R1- R4 respectively map to the sequence of the repair template, while 49.3%, 55%, 35.4%, and 31.1% of reads mapped to the wild type sequence. Red colonies sequenced in flight show heterogeneity. The bar graph depicts the number of reads at each base pair, thus lower coverage corresponds with a lower number of total reads per sample. Nanopore reads are aligned to a hybrid reference sequence that contains both the ADE2 wild type sequence (white bars) and repair template sequence (red bars) at 43–80 bp. S2 Fig: Sequencing coverage plots of red and white colonies from flight and ground controls. Gleason, Conceptualization, Data curation, Methodology, Project administration, Visualization, Writing – original draft, Writing – review & editing, 8, * and Sebastian Kraves, Conceptualization, Formal analysis, Investigation, Methodology, Project administration, Supervision, Writing – original draft, Writing – review & editing 8, * Castro-Wallace, Investigation, Methodology, Writing – review & editing, 7 Ezequiel Alvarez Saavedra, Conceptualization, Methodology, Project administration, Writing – review & editing, 8 Emily J. Scott Copeland, Investigation, Methodology, Writing – review & editing, 6 Sarah L. Foley, Investigation, Methodology, Writing – review & editing, 6 D. Castro, Formal analysis, Visualization, Writing – original draft, Writing – review & editing, 1 Kevin D. Guy Bushkin, Conceptualization, Methodology, Writing – review & editing, # 4, 5 Christian L. Sarah Stahl-Rommel, Data curation, Formal analysis, Investigation, Methodology, Visualization, Writing – original draft, Writing – review & editing, 1 David Li, Conceptualization, Methodology, Writing – review & editing, # 2 Michelle Sung, Conceptualization, Methodology, Writing – review & editing, # 3 Rebecca Li, Conceptualization, Methodology, Writing – review & editing, # 3 Aarthi Vijayakumar, Conceptualization, Methodology, Writing – review & editing, # 3 Kutay Deniz Atabay, Conceptualization, Methodology, Writing – review & editing, # 4, 5 G.
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