Publications

Iwai, Kosuke; sustarich, J.s.; Kim, Peter W.; Gach, Philip C.; Raje, Manasi R.; Heinemann, J.V.H.; Duncombe, Todd A.; Deng, Kai; Northen, T.R.N.; Hillson, N.J.H.; Adams, P.D.A.; Singh, Anup K.

Abstract not provided.

23rd International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2019

Iwai, Kosuke; Wehrs, Maren; Kim, Peter W.; Sustarich, Jess; Northen, Trent R.; Martin, Hector G.; Adams, Paul D.; Singh, Anup K.

We present a novel fully-automated droplet-based microfluidic system to enable programmable combinatorial mixing, electroporation for CRISPR-based gene editing, and high-throughput screening on chip. It is highly robust and compatible with conventional liquid handler systems to interface, enabling 100 different reactions at a time with dramatically lower reagents consumption. Utilizing proposed system, we perform accelerated optimization of biosynthetic pathway of indigoidine in Escherichia coli (E. coli).

Lab on a Chip

Gach, Philip C.; Iwai, Kosuke; Kim, Peter W.; Hillson, Nathan J.; Singh, Anup K.

Synthetic biology is an interdisciplinary field that aims to engineer biological systems for useful purposes. Organism engineering often requires the optimization of individual genes and/or entire biological pathways (consisting of multiple genes). Advances in DNA sequencing and synthesis have recently begun to enable the possibility of evaluating thousands of gene variants and hundreds of thousands of gene combinations. However, such large-scale optimization experiments remain cost-prohibitive to researchers following traditional molecular biology practices, which are frequently labor-intensive and suffer from poor reproducibility. Liquid handling robotics may reduce labor and improve reproducibility, but are themselves expensive and thus inaccessible to most researchers. Microfluidic platforms offer a lower entry price point alternative to robotics, and maintain high throughput and reproducibility while further reducing operating costs through diminished reagent volume requirements. Droplet microfluidics have shown exceptional promise for synthetic biology experiments, including DNA assembly, transformation/transfection, culturing, cell sorting, phenotypic assays, artificial cells and genetic circuits.