Apr 21, 2015

A cell-free framework for biological systems engineering

BioRxiv : the Preprint Server for Biology
Henrike NiederholtmeyerSebastian J Maerkl

Abstract

While complex dynamic biological networks control gene expression and metabolism in all living organisms, engineering comparable synthetic networks remains challenging[1][1],[2][2]. Conducting extensive, quantitative and rapid characterization during the design and implementation process of synthetic networks is currently severely limited due to cumbersome molecular cloning and the difficulties associated with measuring parts, components and systems in cellular hosts. Engineering gene networks in a cell-free environment promises to be an efficient and effective approach to rapidly develop novel biological systems and understand their operating regimes[3][3]-[5][4]. However, it remains questionable whether complex synthetic networks behave similarly in cells and a cell-free environment, which is critical for in vitro approaches to be of significance to biological engineering. Here we show that synthetic dynamic networks can be readily implemented, characterized, and engineered in a cell-free framework and consequently transferred to cellular hosts. We implemented and characterized the “repressilator”[6][5], a three-node negative feedback oscillator in vitro . We then used our cell-free framework to engineer novel three-node, fou...Continue Reading

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Mentioned in this Paper

Metabolic Process, Cellular
In Vivo
Modulation by Virus of Host Cellular Process
Solocyte
Patient Transfer
Environment
Gene Expression
Metabolic Pathway
Organism
Gene Regulatory Networks

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