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Mol Syst Biol. 2011 Mar 1;7:471. doi: 10.1038/msb.2011.4.

A synthetic library of RNA control modules for predictable tuning of gene expression in yeast.

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1
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA, USA.

Abstract

Advances in synthetic biology have resulted in the development of genetic tools that support the design of complex biological systems encoding desired functions. The majority of efforts have focused on the development of regulatory tools in bacteria, whereas fewer tools exist for the tuning of expression levels in eukaryotic organisms. Here, we describe a novel class of RNA-based control modules that provide predictable tuning of expression levels in the yeast Saccharomyces cerevisiae. A library of synthetic control modules that act through posttranscriptional RNase cleavage mechanisms was generated through an in vivo screen, in which structural engineering methods were applied to enhance the insulation and modularity of the resulting components. This new class of control elements can be combined with any promoter to support titration of regulatory strategies encoded in transcriptional regulators and thus more sophisticated control schemes. We applied these synthetic controllers to the systematic titration of flux through the ergosterol biosynthesis pathway, providing insight into endogenous control strategies and highlighting the utility of this control module library for manipulating and probing biological systems.

PMID:
21364573
PMCID:
PMC3094065
DOI:
10.1038/msb.2011.4
[Indexed for MEDLINE]
Free PMC Article
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