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.
Department of Bioengineering, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
Michael S
MS
Magaraci
Department of Bioengineering, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
Jessica G
JG
Bermudez
Department of Bioengineering, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
Deeksha
D
Yogish
Department of Bioengineering, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
Daniel H
DH
Pak
Department of Bioengineering, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
Viktor
V
Mollov
Department of Bioengineering, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
Josh
J
Tycko
Department of Bioengineering, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
David
D
Issadore
Department of Bioengineering, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
Sevile G
SG
Mannickarottu
Department of Bioengineering, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
Brian Y
BY
Chow
D054742:Acyl-Butyrolactones / Q000378:metabolism; D048908:Aliivibrio fischeri / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D053263:Gene Regulatory Networks; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007753:Laboratories; D053038:Quorum Sensing / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D058615:Synthetic Biology / Q000193:education* / Q000379:methods*; D059008:Time-Lapse Imaging / Q000379:methods*; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism
D054742:Acyl-Butyrolactones; D012097:Repressor Proteins; D015534:Trans-Activators; C064778:LuxR autoinducer binding proteins; D049452:Green Fluorescent Proteins
LuxR; STEM; hardware; quorum sensing