Toolbox for Exploring Modular Gene Regulation in Synthetic Biology Training

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.