Development of Cell-Based Sentinels for Nitric Oxide: Ensuring Marker Expression and Unimodality.

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TitleDevelopment of Cell-Based Sentinels for Nitric Oxide: Ensuring Marker Expression and Unimodality.
Publication TypeJournal Article
Year of Publication2018
AuthorsMckay, R, Hauk, P, Quan, D, Bentley, WE
JournalACS Synth Biol
Volume7
Issue7
Pagination1694-1701
Date Published2018 Jul 20
ISSN2161-5063
Abstract

We generated "sentinel" bacteria that respond to the biomarker nitric oxide (NO) and produce a homogeneous and strong fluorescent response. Our dual-plasmid system consists of a signal "relay" vector that employs an NO-responsive promoter that amplifies the native signal (via expression of T7 Polymerase (T7Pol)) to a second vector responsible for GFP expression. Importantly, to achieve an optimal "sentinel" response, we developed strategies that balance the transcriptional load within cells by altering (i) translation and (ii) activity of the T7Pol. Our optimized genetic circuitry was then used to transform commensal E. coli Nissle, as a proof-of-concept toward an ingestible cell-based sensor for Crohn's disease (CD) that, in turn, is marked by elevated levels of intestinal NO. Thus, the "biosensors" demonstrated here may serve as a simple diagnostic tool, contrasting the standard of care including colonoscopies or biopsies.

DOI10.1021/acssynbio.8b00146
Alternate JournalACS Synth Biol
PubMed ID29975512