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

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 Escheri...

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Bibliographic Details
Published in:ACS synthetic biology 2020-01, Vol.9 (1), p.84-94
Main Authors: Liu, Xiangyang, Silverman, Adam D, Alam, Khalid K, Iverson, Erik, Lucks, Julius B, Jewett, Michael C, Raman, Srivatsan
Format: Article
Language:English
Subjects:
Bacterial Proteins - metabolism
Biosensing Techniques - methods
Cell-Free System
Chimera - genetics
Escherichia coli - genetics
Gene Expression
Gene Expression Regulation, Bacterial
Ligands
Plasmids - genetics
Promoter Regions, Genetic
Pseudomonas - genetics
Synthetic Biology - methods
Transcription Factors - metabolism
Transcription, Genetic
Triazines - analysis
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Summary: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.
ISSN:2161-5063
2161-5063
DOI:10.1021/acssynbio.9b00348