Date of this Version
ACS Synthetic Biology
We report a toolbox for exploring the modular tuning of genetic circuits, which has been specifically optimized for widespread deployment in STEM environments through a combination of bacterial strain engineering and distributable hardware development. The transfer functions of 16 genetic switches, programmed to express a GFP reporter under the regulation of the (acyl-homoserine lactone) AHL-sensitive luxR transcriptional activator, can be parametrically tuned by adjusting high/low degrees of transcriptional, translational, and post-translational processing. Strains were optimized to facilitate daily large-scale preparation and reliable performance at room temperature in order to eliminate the need for temperature controlled apparatuses, which are both cost-limiting and space-constraining. The custom-designed, automated, and web-enabled fluorescence documentation system allows time-lapse imaging of AHL-induced GFP expression on bacterial plates with real-time remote data access, thereby requiring trainees to only be present for experimental setup. When coupled with mathematical models in agreement with empirical data, this toolbox expands the scalability and scope of reliable synthetic biology experiments for STEM training.
This document is the unedited author's version of a Submitted Work that was subsequently accepted for publication in ACS Synthetic Biology, copyright © American Chemical Society after peer review. To access the final edited and published work, see http://dx.doi.org/10.1021/acssynbio.6b00057.
STEM, quorum sensing, LuxR, hardware
Magaraci, M. S., Bermudez, J. G., Yogish, D., Pak, D. H., Mollov, V., Tycko, J., Issadore, D., Mannickarottu, S. G., & Chow, B. Y. (2016). Toolbox for Exploring Modular Gene Regulation in Synthetic Biology Training. ACS Synthetic Biology, http://dx.doi.org/10.1021/acssynbio.6b00057
Additional FilesCAD Files.zip (242174 kB)
Raspberry Pi OS.img (5046272 kB)
Sample Protocols.pdf (68 kB)
User Manual.pdf (8676 kB)
Date Posted: 08 July 2016
This document has been peer reviewed.