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Genetically Programmable Microbial Assembly

Kozlowski, Mark T. and Silverman, Bradley R. and Johnstone, Christopher P. and Tirrell, David A. (2021) Genetically Programmable Microbial Assembly. ACS Synthetic Biology, 10 (6). pp. 1351-1359. ISSN 2161-5063. PMCID PMC8978479. doi:10.1021/acssynbio.0c00616.

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Engineered microbial communities show promise in a wide range of applications, including environmental remediation, microbiome engineering, and synthesis of fine chemicals. Here we present methods by which bacterial aggregates can be directed into several distinct architectures by inducible surface expression of heteroassociative protein domains (SpyTag/SpyCatcher and SynZip17/18). Programmed aggregation can be used to activate a quorum-sensing circuit, and aggregate size can be tuned via control of the amount of the associative protein displayed on the cell surface. We further demonstrate reversibility of SynZip-mediated assembly by addition of soluble competitor peptide. Genetically programmable bacterial assembly provides a starting point for the development of new applications of engineered microbial communities in environmental technology, agriculture, human health, and bioreactor design.

Item Type:Article
Related URLs:
URLURL TypeDescription CentralArticle
Silverman, Bradley R.0000-0002-9256-8941
Johnstone, Christopher P.0000-0002-7255-0218
Tirrell, David A.0000-0003-3175-4596
Additional Information:© 2021 American Chemical Society. Received: December 7, 2020; Published: May 19, 2021. This work was supported by the Defense Advanced Research Projects Agency under award numbers HR0011-15-C-0093 and HR0011-17-2-0037. M.T.K. was supported by the Department of Defense (DoD) through the National Defense Science & Engineering Graduate Fellowship (NDSEG) Program. B.R.S. was supported by NIH Training Grant 1T32GM112592. C.P.J. was supported by a Caltech Summer Undergraduate Research Fellowship. Plasmid pHEA-Cm, from which the autotransporter was cloned, was a kind gift of Luis Angel Fernandez Herrero of the Centro Nacional de Biotecnologia (Madrid, Spain). Plasmid pLuxRI2, from which we cloned the quorum-sensing plasmid, was a gift of Frances Arnold of the California Institute of Technology (Pasadena, California). pKPY680, pKPY681, and DH10B strains KY35 and KY36 were prepared by Kai P. Yuet. We thank Dr. Andres Collazo for assistance with confocal microscopy. Imaging was performed in the Biological Imaging Facility of the Caltech Beckman Institute, which is supported by the Arnold and Mabel Beckman Foundation. We thank Yaron Antebi for use of EasyFlow for flow cytometry analysis, and Adam Silverman for a critical review of the manuscript. Author Contributions: M.T.K. and B.R.S. contributed equally to this work. Author Contributions: M.T.K., B.R.S., and D.A.T. conceived the study. M.T.K., B.R.S., and C.P.J. conducted all the experiments in the paper. B.R.S. wrote computational tools for image analysis and reaction diffusion modeling. M.T.K., B.R.S., and D.A.T. wrote the paper. All authors read and approved the final manuscript. The authors declare no competing financial interest.
Funding AgencyGrant Number
Defense Advanced Research Projects Agency (DARPA)HR0011-15-C-0093
Defense Advanced Research Projects Agency (DARPA)HR0011-17-2-0037
National Defense Science and Engineering Graduate (NDSEG) FellowshipUNSPECIFIED
NIH Predoctoral Fellowship1T32GM112592
National Institute of General Medical SciencesUNSPECIFIED
Caltech Summer Undergraduate Research Fellowship (SURF)UNSPECIFIED
Arnold and Mabel Beckman FoundationUNSPECIFIED
Subject Keywords:Quorum sensing, Bacteria, Protein dynamics, Aggregation, Electrochemical cells
Issue or Number:6
PubMed Central ID:PMC8978479
Record Number:CaltechAUTHORS:20210519-141320901
Persistent URL:
Official Citation:Genetically Programmable Microbial Assembly. Mark T. Kozlowski, Bradley R. Silverman, Christopher P. Johnstone, and David A. Tirrell. ACS Synthetic Biology 2021 10 (6), 1351-1359; DOI: 10.1021/acssynbio.0c00616
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:109192
Deposited By: George Porter
Deposited On:24 May 2021 14:37
Last Modified:12 Jul 2022 18:19

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