a modified consumer inkjet for spatiotemporal control of gene expression修改后的消费者喷墨时空控制基因的表达.pdfVIP

A Modified Consumer Inkjet for Spatiotemporal Control of Gene Expression 1 2 3 Daniel J. Cohen *, Roberto C. Morfino , Michel M. Maharbiz ´ 1 Department of Bioengineering, University of California, Berkeley, California, United States of America, 2 Department of Electrical Engineering, Ecole Polytechnique ´ ´ Federale de Lausanne, Lausanne, Switzerland, 3 Department of Electrical Engineering and Computer Science, University of California, Berkeley, California, United States of America Abstract 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 mm 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 ope