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Controlled exchange of protein and nucleic acid signals from and between synthetic minimal cells

Joseph M. Heili, Kaitlin Stokes, Nathaniel J. Gaut, Christopher Deich, Jose Gomez-Garcia, Brock Cash, Matthew R. Pawlak, Aaron E Engelhart, Katarzyna P. Adamala
doi: https://doi.org/10.1101/2022.01.03.474826
Joseph M. Heili
1Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN US
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Kaitlin Stokes
1Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN US
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Nathaniel J. Gaut
1Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN US
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Christopher Deich
1Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN US
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Jose Gomez-Garcia
1Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN US
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Brock Cash
1Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN US
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Matthew R. Pawlak
1Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN US
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Aaron E Engelhart
1Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN US
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Katarzyna P. Adamala
1Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN US
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  • For correspondence: kadamala@umn.edu
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Abstract

Synthetic minimal cells are a class of small liposome bioreactors that have some, but not all functions of live cells. Here, we report a critical step towards the development of a bottom-up minimal cell: cellular export of functional protein and RNA products. We used cell penetrating peptide tags to translocate payloads across a synthetic cell vesicle membrane. We demonstrated efficient transport of active enzymes, and transport of nucleic acid payloads by RNA binding proteins. We investigated influence of a concentration gradient alongside other factors on the efficiency of the translocation, and we show a method to increase product accumulation in one location. We demonstrate the use of this technology to engineer molecular communication between different populations of synthetic cells, to exchange protein and nucleic acid signals. The synthetic minimal cell production and export of proteins or nucleic acids allows experimental designs that approach the complexity and relevancy of natural biological systems.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted January 03, 2022.
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Controlled exchange of protein and nucleic acid signals from and between synthetic minimal cells
Joseph M. Heili, Kaitlin Stokes, Nathaniel J. Gaut, Christopher Deich, Jose Gomez-Garcia, Brock Cash, Matthew R. Pawlak, Aaron E Engelhart, Katarzyna P. Adamala
bioRxiv 2022.01.03.474826; doi: https://doi.org/10.1101/2022.01.03.474826
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Controlled exchange of protein and nucleic acid signals from and between synthetic minimal cells
Joseph M. Heili, Kaitlin Stokes, Nathaniel J. Gaut, Christopher Deich, Jose Gomez-Garcia, Brock Cash, Matthew R. Pawlak, Aaron E Engelhart, Katarzyna P. Adamala
bioRxiv 2022.01.03.474826; doi: https://doi.org/10.1101/2022.01.03.474826

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