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Tunable integrase-mediated differentiation facilitates improved output of burdensome functions in E. coli

Williams, Rory L. and Murray, Richard M. (2019) Tunable integrase-mediated differentiation facilitates improved output of burdensome functions in E. coli. . (Unpublished) http://resolver.caltech.edu/CaltechAUTHORS:20190422-091140053

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Abstract

Application of synthetic biology is limited by the capacity of cells to faithfully execute burdensome engineered functions in the face of Darwinian evolution. Division of labor, both metabolic and reproductive, are underutilized in confronting this barrier. To address this, we developed a serine-integrase based differentiation circuit that allows control of the population composition through tuning of the differentiation rate and number of cell divisions differentiated cells can undergo. We applied this system to T7 RNAP-driven expression of a fluorescent protein, and demonstrate both increased duration of circuit function and total production for high burden expression. While T7 expression systems are typically used for high-level short-term expression, this system enables longer duration production, and could be readily applied to burdensome or toxic products not readily produced in bacteria.


Item Type:Report or Paper (Discussion Paper)
Related URLs:
URLURL TypeDescription
https://doi.org/10.1101/614529DOIDiscussion Paper
https://www.biorxiv.org/content/10.1101/614545v1.supplementary-materialPublisherSupporting Information
ORCID:
AuthorORCID
Williams, Rory L.0000-0003-2605-5790
Murray, Richard M.0000-0002-5785-7481
Additional Information:The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. This article is a US Government work. It is not subject to copyright under 17 USC 105 and is also made available for use under a CC0 license. We would like to thank Andy Halleran, Anandh Swaminathan, and Andrey Shur for productive conversations; Andy Halleran for providing code for analysis of flow cytometry data; Samuel Clamons for providing code for tidying and analyzing Biotek data; and Andrey Shur and Andy Halleran for providing cloning resources. pSal, pLas, and pTac and their associated evolved transcription factors were kind gifts from Adam Meyer. The CIDAR MoClo Parts Kit was a gift from Douglas Densmore (Addgene kit # 1000000059). This research is supported by the Institute for Collaborative Biotechnologies through grant W911NF-09-0001 and cooperative agreement W911NF-19-2-0026 from the U.S. Army Research Office. The content of the information on this page does not necessarily reflect the position or the policy of the Government, and no official endorsement should be inferred.
Funders:
Funding AgencyGrant Number
Army Research Office (ARO)W911NF-09-0001
Army Research Office (ARO)W911NF-19-2-0026
Record Number:CaltechAUTHORS:20190422-091140053
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20190422-091140053
Official Citation:Tunable integrase-mediated differentiation facilitates improved output of burdensome functions in E. coli Rory L. Williams, Richard M. Murray bioRxiv 614529; doi: https://doi.org/10.1101/614529
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:94829
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:22 Apr 2019 22:59
Last Modified:22 Apr 2019 22:59

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