A modified consumer inkjet for spatiotemporal control of gene expression.
This paper presents a low-cost inkjet dosing system capable of continuous, two-dimensional spatiotemporal regulation of gene expression via delivery of diffusible regulators to a custom-mounted gel culture of E. coli. A consumer-grade, inkjet printer was adapted for chemical printing; E. coli cultures were grown on 750 microm thick agar embedded in micro-wells machined into commercial compact discs. Spatio-temporal regulation of the lac operon was demonstrated via the printing of patterns of lactose and glucose directly into the cultures; X-Gal blue patterns were used for visual feedback. We demonstrate how the bistable nature of the lac operon's feedback, when perturbed by patterning lactose (inducer) and glucose (inhibitor), can lead to coordination of cell expression patterns across a field in ways that mimic motifs seen in developmental biology. Examples of this include sharp boundaries and the generation of traveling waves of mRNA expression. To our knowledge, this is the first demonstration of reaction-diffusion effects in the well-studied lac operon. A finite element reaction-diffusion model of the lac operon is also presented which predicts pattern formation with good fidelity.
Cohen
Daniel J
DJ
Department of Bioengineering, University of California, Berkeley, California, USA. dancohen@berkeley.edu
Morfino
Roberto C
RC
Maharbiz
Michel M
MM
D020816:Amino Acid Motifs; D004058:Diffusion; D004867:Equipment Design; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics; D020869:Gene Expression Profiling / Q000295:instrumentation* / Q000379:methods*; D015964:Gene Expression Regulation, Bacterial; D005947:Glucose / Q000378:metabolism; D007763:Lac Operon; D007785:Lactose / Q000378:metabolism; D008954:Models, Biological; D011327:Printing / Q000295:instrumentation* / Q000379:methods*; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors
D029968:Escherichia coli Proteins; D012333:RNA, Messenger; D005947:Glucose; D007785:Lactose