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 language | English (US) |
|---|---|
| Pages (from-to) | 84-94 |
| Number of pages | 11 |
| Journal | ACS synthetic biology |
| Volume | 9 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jan 17 2020 |
Funding
The authors would like to acknowledge members of the Raman, Jewett, and Lucks laboratories for helpful discussions. S.R. and X.L. were supported by the University of Wisconsin–Madison and Army Research Office grant W911NF1710043. M.C.J. acknowledges support from the Air Force Research Laboratory Center of Excellence Grant FA8650-15-2-5518, the DARPA 1000 Molecules Program HR0011-15-C-0084, the David and Lucile Packard Foundation, and the Camille Dreyfus Teacher-Scholar Program. J.B.L. acknowledges support from the Air Force Research Laboratory Center of Excellence Grant FA8650-15-2-5518, the Camille Dreyfus Teacher-Scholar Program and Searle Funds at the Chicago Community Trust. A.D.S. was supported in part by the National Institutes of Health Training Grant (T32GM008449) through Northwestern University’s Biotechnology Training Program. The views and conclusions contained herein are those of the authors and should not be interpreted as necessarily representing the official policies or endorsements, either expressed or implied, of the Army Research Office, Air Force Research Laboratory, Air Force Office of Scientific Research, DARPA, or the U.S. Government.
ASJC Scopus subject areas
- Biomedical Engineering
- Biochemistry, Genetics and Molecular Biology (miscellaneous)
