PT - JOURNAL ARTICLE AU - Andrew D. Halleran AU - Richard M. Murray TI - Cell-free and <em>in vivo</em> characterization of Lux, Las, and Rpa quorum activation systems in <em>E. coli</em> AID - 10.1101/159988 DP - 2017 Jan 01 TA - bioRxiv PG - 159988 4099 - http://biorxiv.org/content/early/2017/07/07/159988.short 4100 - http://biorxiv.org/content/early/2017/07/07/159988.full AB - Synthetic biologists have turned towards 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.