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  Cell-free gene expression dynamics in synthetic cell populations

Gonzales, D. T., Yandrapalli, N., Robinson, T., Zechner, C., & Tang, T.-Y.-D. (2022). Cell-free gene expression dynamics in synthetic cell populations. ACS Synthetic Biology, 11(1), 205-215. doi:10.1021/acssynbio.1c00376.

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Item Permalink: https://hdl.handle.net/21.11116/0000-0009-C85F-C Version Permalink: https://hdl.handle.net/21.11116/0000-000B-04FC-5
Genre: Journal Article

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 Creators:
Gonzales, David T., Author
Yandrapalli, Naresh1, Author           
Robinson, Tom1, Author           
Zechner, Christoph, Author
Tang, T-Y. Dora, Author
Affiliations:
1Tom Robinson, Theorie & Bio-Systeme, Max Planck Institute of Colloids and Interfaces, Max Planck Society, ou_2288691              

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 Abstract: The ability to build synthetic cellular populations from the bottom-up provides the groundwork to realize minimal living tissues comprising single cells which can communicate and bridge scales into multicellular systems. Engineered systems made of synthetic micron-sized compartments and integrated reaction networks coupled with mathematical modeling can facilitate the design and construction of complex and multiscale chemical systems from the bottom-up. Toward this goal, we generated populations of monodisperse liposomes encapsulating cell-free expression systems (CFESs) using double-emulsion microfluidics and quantified transcription and translation dynamics within individual synthetic cells of the population using a fluorescent Broccoli RNA aptamer and mCherry protein reporter. CFE dynamics in bulk reactions were used to test different coarse-grained resource-limited gene expression models using model selection to obtain transcription and translation rate parameters by likelihood-based parameter estimation. The selected model was then applied to quantify cell-free gene expression dynamics in populations of synthetic cells. In combination, our experimental and theoretical approaches provide a statistically robust analysis of CFE dynamics in bulk and monodisperse synthetic cell populations. We demonstrate that compartmentalization of CFESs leads to different transcription and translation rates compared to bulk CFE and show that this is due to the semipermeable lipid membrane that allows the exchange of materials between the synthetic cells and the external environment.

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Language(s): eng - English
 Dates: 2022-01-042022
 Publication Status: Issued
 Pages: -
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 Rev. Type: -
 Identifiers: DOI: 10.1021/acssynbio.1c00376
BibTex Citekey: doi:10.1021/acssynbio.1c00376
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Title: ACS Synthetic Biology
  Abbreviation : ACS Synth. Biol.
Source Genre: Journal
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Publ. Info: Washington, D.C. : American Chemical Society
Pages: - Volume / Issue: 11 (1) Sequence Number: - Start / End Page: 205 - 215 Identifier: ISSN: 2161-5063