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Published March 2011 | Published + Supplemental Material
Journal Article Open

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


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.

Additional Information

© 2011 EMBO and Macmillan Publishers Limited. This is an open-access article distributed under the terms of the Creative Commons Attribution Noncommercial No Derivative Works 3.0 Unported License, which permits distribution and reproduction in any medium, provided the original author and source are credited. This license does not permit commercial exploitation or the creation of derivative works without specific permission. Received 30 August 2010; Accepted 18 January 2011; Published online 1 March 2011. We thank K Hoff, S Bastian and F.H. Arnold for assistance in the purification of Rnt1p and J Liang for assistance with in vitro assays. This work was supported by the National Science Foundation (CAREER award to CDS; CBET-0917705) and the Alfred P Sloan Foundation (fellowship to CDS). Author contributions: AHB designed research, performed research and wrote the paper; CDS designed research and wrote the paper.

Attached Files

Published - Babiskin2011p13554Mol_Syst_Biol.pdf

Supplemental Material - msb20114-df1D.xls

Supplemental Material - msb20114-df3A.xls

Supplemental Material - msb20114-df3B.xls

Supplemental Material - msb20114-df3C.xls

Supplemental Material - msb20114-df4A.xls

Supplemental Material - msb20114-df4B.xls

Supplemental Material - msb20114-df5B.xls

Supplemental Material - msb20114-df5D.xls

Supplemental Material - msb20114-df6C.xls

Supplemental Material - msb20114-df6D.xls

Supplemental Material - msb20114-df6E.xls

Supplemental Material - msb20114-s1.pdf


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August 22, 2023
October 23, 2023