Cell-free synthetic biology in nanofabricated reaction devices

David Karig (October 27, 2011)

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Abstract

The growing field of synthetic biology aims to forward engineer biology both for applications such as energy production, drug production, and bioremediation, as well as for the purpose of furthering the fundamental understanding of natural systems. However, engineering living cells is notoriously difficult due to issues such as mutation, epigenetic variation, fitness effects, and the interaction of synthetic components with host cell processes. Thus, simpler contexts such as cell-free expression systems offer great promise to engineering complex biological behavior in a quantitative fashion. Furthermore, the confinement of cell-free gene circuit reactions in nanofabricated reaction devices offers a flexible approach to investigating fundamental aspects of gene circuit function. Currently, we are using such devices to study noise in simple gene circuits. Cell-free reactions confined in different volume wells are imaged over time using fluorescent microscopy. The noise characteristics of the resulting gene expression trajectories are analyzed and compared for different gene circuits.