Validation of cell-free protein synthesis aboard the International Space Station

Kocalar, S.; Miller, B. M.; Huang, A.; Gleason, E.; Martin, K.; Foley, K.; Copeland, D. S.; Jewett, M.; Saavedra, E. A.; Kraves, S.

doi:10.1101/2023.12.06.570403

bioRxiv · 10.1101/2023.12.06.570403

Validation of cell-free protein synthesis aboard the International Space Station

Kocalar, S.·Miller, B. M.·Huang, A.·Gleason, E.·Martin, K.·Foley, K.·Copeland, D. S.·Jewett, M.·Saavedra, E. A.·Kraves, S.

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Abstract

Cell-free protein synthesis (CFPS) is a rapidly maturing in vitro gene expression platform that can be used to transcribe and translate nucleic acids at the point of need, enabling on-demand synthesis of peptide-based vaccines and biotherapeutics, as well as the development of diagnostic tests for environmental contaminants and infectious agents. Unlike traditional cell-based systems, CFPS platforms do not require the maintenance of living cells and can be deployed with minimal equipment; therefore, they hold promise for applications in low-resource contexts, including spaceflight. Here we evaluate the performance of cell-free BioBits(R) platform aboard the International Space Station by expressing RNA-based aptamers and fluorescent proteins that can serve as biological indicators. We validate two classes of biological sensors that detect either the small molecule DFHBI or a specific RNA sequence. Upon detection of their respective analytes, both biological sensors produce fluorescent readouts that are visually confirmed using a handheld fluorescence viewer and imaged for quantitative analysis. Our findings provide insight into the kinetics of cell-free transcription and translation in a microgravity environment and reveal that both biosensors perform robustly in space. Our findings lay the groundwork for portable, low-cost applications ranging from point-of-care health monitoring to on-demand detection of environmental hazards in low-resource communities both on Earth and beyond. Visual graphic O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=123 SRC="FIGDIR/small/570403v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@140f8f2org.highwire.dtl.DTLVardef@97dda3org.highwire.dtl.DTLVardef@5824bforg.highwire.dtl.DTLVardef@d47ded_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Kocalar, S., Miller, B. M., Huang, A., Gleason, E., Martin, K., Foley, K., Copeland, D. S., Jewett, M., Saavedra, E. A., Kraves, S.. 2023-12-06. Validation of cell-free protein synthesis aboard the International Space Station. https://doi.org/10.1101/2023.12.06.570403

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