CaltechAUTHORS
  A Caltech Library Service

Cell-Free and In Vivo Characterization of Lux, Las, and Rpa Quorum Activation Systems in E. coli

Halleran, Andrew D. and Murray, Richard M. (2018) Cell-Free and In Vivo Characterization of Lux, Las, and Rpa Quorum Activation Systems in E. coli. ACS Synthetic Biology, 7 (2). pp. 752-755. ISSN 2161-5063. http://resolver.caltech.edu/CaltechAUTHORS:20171204-081929055

[img] PDF - Submitted Version
Creative Commons Attribution Non-commercial No Derivatives.

7Mb
[img] PDF (Complete Materials and Methods; Supporting Table S1, containing all sequence information for constructs used; Supporting Figures S1-S33) - Supplemental Material
See Usage Policy.

2743Kb

Use this Persistent URL to link to this item: http://resolver.caltech.edu/CaltechAUTHORS:20171204-081929055

Abstract

Synthetic biologists have turned toward quorum systems as a path for building sophisticated microbial consortia that exhibit group decision making. Currently, however, even the most complex consortium circuits rely on only one or two quorum sensing systems, greatly restricting the available design space. High-throughput characterization of available quorum sensing systems is useful for finding compatible sets of systems that are suitable for a defined circuit architecture. Recently, cell-free systems have gained popularity as a test-bed for rapid prototyping of genetic circuitry. We take advantage of the transcription–translation cell-free system to characterize three commonly used Lux-type quorum activators, Lux, Las, and Rpa. We then compare the cell-free characterization to results obtained in vivo. We find significant genetic crosstalk in both the Las and Rpa systems and substantial signal crosstalk in Lux activation. We show that cell-free characterization predicts crosstalk observed in vivo.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1021/acssynbio.7b00376DOIArticle
http://pubs.acs.org/doi/abs/10.1021/acssynbio.7b00376PublisherArticle
http://pubs.acs.org/doi/suppl/10.1021/acssynbio.7b00376PublisherSupporting Information
http://dx.doi.org/10.1101/159988DOIDiscussion Paper
ORCID:
AuthorORCID
Halleran, Andrew D.0000-0001-8720-1451
Murray, Richard M.0000-0002-5785-7481
Additional Information:© 2017 American Chemical Society. Received: October 16, 2017; Published: November 9, 2017. The authors would like to thank Sam Clamons, Andrey Shur and Vipul Singhal for fruitful discussions. We also thank John Marken for helpful comments on the manuscript. Plasmid vectors were provided by Douglas Densmore at the Cross-disciplinary Integration of Design Automation Research lab (Addgene Kit # 1000000059). The project depicted was sponsored by the Defense Advanced Research Projects Agency (Agreement HR0011-17-2-0008). The content of the information does not necessarily reflect the position or the policy of the Government, and no official endorsement should be inferred. The authors declare the following competing financial interest(s): RMM has ownership in a company that commercializes the cell-free system used in this manuscript.
Funders:
Funding AgencyGrant Number
Defense Advanced Research Projects Agency (DARPA)HR0011-17-2-0008
Subject Keywords:cell−cell communication, quorum systems, cell-free, population circuitry
Record Number:CaltechAUTHORS:20171204-081929055
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20171204-081929055
Official Citation:Cell-Free and In Vivo Characterization of Lux, Las, and Rpa Quorum Activation Systems in E. coli. Andrew D. Halleran and Richard M. Murray. ACS Synthetic Biology 2018 7 (2), 752-755. DOI: 10.1021/acssynbio.7b00376
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:83654
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:13 Dec 2017 05:11
Last Modified:01 Mar 2018 23:14

Repository Staff Only: item control page