Engineering Platforms for Advancing Plant Synthetic Biology

This work describes research aimed at adapting advanced engineering systems for plant biotechnology. The droplet interface bilayer (DIB) is a robust and versatile platform for replicating model cell membranes, providing a bottom-up approach for synthetic cell- and tissue-like structures. In this work, a microfluidic device featuring five inlets, one for the continuous oil phase and four discrete aqueous channels for droplet generation was designed. Droplet production rates were controlled by adjusting the applied pressure of each inlet; and thus, altering the droplet sequence for capturing linear DIB networks in a downstream hydrodynamic trapping array. This microfluidic system provides a high-throughput method for generating DIB networks of complex droplet patterning for the purpose of studying higher-order combinatorial networks and signal cascades. Further, this work characterizes pressure-driven flow sensitivity to changes in microchannel resistance and differences in hydrodynamic trapping behavior compared to mechanically-driven flow.

In addition, this research work developed a custom laser system to image plant canopies expressing a wide variety of fluorescent protein genes in leaves for the purpose of phenotyping transgenic plants expressing multiple constitutive or inducible fluorescent proteins. The fluorescence-inducing laser projector (FILP) system is a critical instrument for accessing the efficacy of environmental phytosensors, which are plants engineered to detect and report environmental stimuli.

Portions of this document were previously published in Plant Biotechnology Journal:

Tayler M. Schimel, Mary-Anne Nguyen, Stephen A. Sarles, Scott C. Lenaghan. “Pressure-driven generation of complex microfluidic droplet networks.” (2020).

Stephen B. Rigoulot, Tayler M. Schimel, Jun Hyung Lee, Robert G. Sears, Holly Brabazon, Jessica S. Layton, Li Li, Kerry A. Meier, Magen R. Poindexter, Manuel J. Schmid, Erin M. Seaberry, Jared W. Brabazon, Jonathan A. Madajian, Michael J. Finander, John DiBenedetto, Alessandro Occhialini, Scott C. Lenaghan, C. Neal Stewart, Jr. "Imaging of multiple fluorescent proteins in canopies enables synthetic biology in plants." Plant Biotechnology Journal (2020).