Design of a Transcriptional Biosensor for the Portable, On-Demand Detection of Cyanuric Acid
| Autor: | Srivatsan Raman, Adam D. Silverman, Khalid K. Alam, Erik Iverson, Michael C. Jewett, Xiangyang Liu, Julius B. Lucks |
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| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
Analyte Transcription Genetic Biomedical Engineering Gene Expression Context (language use) Nanotechnology Biosensing Techniques Computational biology Ligands medicine.disease_cause 01 natural sciences Biochemistry Genetics and Molecular Biology (miscellaneous) Article 03 medical and health sciences chemistry.chemical_compound Synthetic biology Bacterial Proteins Pseudomonas 010608 biotechnology On demand Escherichia coli medicine Promoter Regions Genetic 030304 developmental biology 0303 health sciences Cell-Free System Chimera Triazines 030306 microbiology Extramural Promoter Gene Expression Regulation Bacterial General Medicine chemistry Synthetic Biology Cyanuric acid Biosensor DNA Function (biology) Plasmids Transcription Factors |
| Zdroj: | ACS Synth Biol |
| ISSN: | 2161-5063 |
| DOI: | 10.1021/acssynbio.9b00348 |
| Popis: | 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 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. |
| Databáze: | OpenAIRE |
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