Design of a Transcriptional Biosensor for the Portable, On-Demand Detection of Cyanuric Acid

Xiangyang Liu, Adam D. Silverman, Khalid K. Alam, Erik Iverson, Julius B. Lucks*, Michael C. Jewett, Srivatsan Raman

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

Rapid molecular biosensing is an emerging application area for synthetic biology. Here, we engineer a portable biosensor for cyanuric acid (CYA), an analyte of interest for human and environmental health, using a LysR-type transcription regulator (LTTR) from Pseudomonas within the context of Escherichia coli gene expression machinery. To overcome cross-host portability challenges of LTTRs, we rationally engineered hybrid Pseudomonas-E. coli promoters by integrating DNA elements required for transcriptional activity and ligand-dependent regulation from both hosts, which enabled E. coli to function as a whole-cell biosensor for CYA. To alleviate challenges of whole-cell biosensing, we adapted these promoter designs to function within a freeze-dried E. coli cell-free system to sense CYA. This portable, on-demand system robustly detects CYA within an hour from laboratory and real-world samples and works with both fluorescent and colorimetric reporters. This work elucidates general principles to facilitate the engineering of a wider array of LTTR-based environmental sensors.

Original languageEnglish (US)
JournalACS synthetic biology
DOIs
StateAccepted/In press - Jan 1 2020

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ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

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