Detection of DNA Microsatellites Using Multiplex Polymerase Chain Reaction Aboard the International Space Station

Sophia Chen, J. Hatch, A. Luck, N. Nichols, Emily J Gleason, K. Martin, Kevin D. Foley, D. Scott Copeland, Sebastian Kraves, E. Saavedra
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Abstract

Abstract As human exploration extends further into deep space, it is critical to understand the cellular impacts of spaceflight in order to ensure the safety of future astronauts. Extended exposure to cosmic radiation and microgravity has been shown to cause genetic damage and impair cellular DNA repair mechanisms, which together can lead to genomic instability. In particular, microsatellite instability (MSI), in which dysfunction in DNA mismatch repair (MMR) causes alterations in tandemly repeated “microsatellite” sequences, is a manifestation of genomic instability that has been associated with certain cancers. In this study, we establish the feasibility of an on-orbit multiplex polymerase chain reaction (PCR)-based assay to detect mutations in cancer-related microsatellites. Multiplex PCR was used to amplify five quasimonomorphic microsatellites in space and on Earth from both wild-type and MMR-deficient human cell lines. These data provide proof of concept of simultaneous amplification of multiple DNA sequences in space, expanding in-flight research and health-monitoring capabilities.
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利用多重聚合酶链反应在国际空间站上检测DNA微卫星
随着人类探索进一步深入太空,了解太空飞行对细胞的影响对于确保未来宇航员的安全至关重要。长期暴露在宇宙辐射和微重力下已被证明会造成遗传损伤和损害细胞DNA修复机制,两者共同导致基因组不稳定。特别是,微卫星不稳定性(MSI),即DNA错配修复(MMR)功能障碍导致串联重复的“微卫星”序列改变,是与某些癌症相关的基因组不稳定性的一种表现。在这项研究中,我们建立了一种基于在轨多重聚合酶链反应(PCR)的检测癌症相关微卫星突变的可行性。利用多重PCR技术,从野生型和mmr缺陷的人类细胞系中扩增出5个准单态微卫星。这些数据证明了在太空中同时扩增多个DNA序列的概念,扩大了飞行中研究和健康监测能力。
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