Abstract

New Generation of Droplet Technology for Attoliter- Scale Synthetic Biology

Manish Biyani, Associate Professor, Japan Advanced Institute of Science and Technology

The bottom-up construction of artificial cells using IVC (in vitro compartmentalization) technology in synthetic biology has redefined the efficiency of cell-free system. The production rate of cell-free reaction has realized to be inversely proportional to the reaction-scale size. This is mainly because bulk-scale reaction (ranging from milliliters-to-microliter scale) in unconfined spaces leads to noise due to non-Poisson distribution of reaction components. Second, compartmentalization induces cell-like interfacial phenomenon by separating the inner vital chemical milieu from external environment, which increases with decreasing sizes of compartment. Therefore, a method of IVC that can perform cell-free reactions in confined space (ranging from femtoliter-to-attoliter scale) is an exciting prospect in synthetic biology.

This talk will highlight a recently developed new electrospray platform for minimal solute entrapment in femtoliter droplets for noise-free synthetic biology [Sci. Rep. 2016:6,26257]. Next, a combination of droplet technology and cell-free reaction enables to encapsulate biology in a cell-sized confined space and thus offers a means to parallelize biological and chemical assays inside of femtoliter-sized microdroplet compartments for in vitro molecular evolution and selection. This system is capable of continuous and rapid generation of large-scale and highly monodisperse cell-like compartments with volume <5 fL at the speed of 3×106 per min using the application of electrostatic super-fine inkjet technology. We further extended the utility of this system by introducing a micro-hole array electrospray platform for ultra-large scale generation of attoliter-scale cell-like compartments. These droplets were utilized to perform in vitro biological reactions and further exploited for in vitro selection of highly functional biomolecules.