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Cell-Free and In Vivo Characterization of Lux, Las, and Rpa Quorum Activation Systems in E. coli
journal contribution
posted on 2017-11-09, 00:00 authored by Andrew D. Halleran, Richard M. MurraySynthetic
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.