Steady-State Cell-Free Gene Expression with Microfluidic Chemostats

Nadanai Laohakunakorn; Barbora Lavickova; Zoe Swank; Julie Laurent; Sebastian J Maerkl

doi:10.1007/978-1-0716-1032-9_9

Steady-State Cell-Free Gene Expression with Microfluidic Chemostats

Methods Mol Biol. 2021:2229:189-203. doi: 10.1007/978-1-0716-1032-9_9.

Authors

Nadanai Laohakunakorn¹, Barbora Lavickova², Zoe Swank², Julie Laurent², Sebastian J Maerkl³

Affiliations

¹ School of Biological Sciences, Institute of Quantitative Biology, Biochemistry and Biotechnology, University of Edinburgh, Edinburgh, UK.
² Institute of Bioengineering, School of Engineering, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
³ Institute of Bioengineering, School of Engineering, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland. sebastian.maerkl@epfl.ch.

PMID: 33405223
DOI: 10.1007/978-1-0716-1032-9_9

Abstract

Cell-free synthetic biology offers an approach to building and testing gene circuits in a simplified environment free from the complexity of a living cell. Recent advances in microfluidic devices allowed cell-free reactions to run under nonequilibrium, steady-state conditions enabling the implementation of dynamic gene regulatory circuits in vitro. In this chapter, we present a detailed protocol to fabricate a microfluidic chemostat device which enables such an operation, detailing essential steps in photolithography, soft lithography, and hardware setup.

Keywords: Cell-free; Microfluidics; Steady-state gene expression; Synthetic biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell-Free System
Gene Expression
Gene Regulatory Networks*
Humans
Microfluidic Analytical Techniques / instrumentation*
Software
Synthetic Biology / methods*