Issue 8, 2011

Selection of tetracycline inducible self-cleaving ribozymes as synthetic devices for gene regulation in yeast

Abstract

Synthetic regulatory devices are key components for the development of complex biological systems and the reprogramming of cellular functions and networks. Here we describe the selection of tetracycline inducible hammerhead ribozymes. A tetracycline aptamer was fused to the full-length hammerhead ribozymevia a variable linker region. 11 rounds of in vitro selection were applied to isolate linker sequences that mediate tetracycline dependent hammerhead cleavage. We identified allosteric ribozymes that cleave in the presence of 1 μM tetracycline as fast as the full-length ribozyme whereas cleavage is inhibited up to 333-fold in the absence of tetracycline. Reporter gene assays indicate that the allosteric ribozymes can be employed to control gene expression in yeast.

Graphical abstract: Selection of tetracycline inducible self-cleaving ribozymes as synthetic devices for gene regulation in yeast

Supplementary files

Article information

Article type
Paper
Submitted
21 Feb 2011
Accepted
04 May 2011
First published
20 May 2011

Mol. BioSyst., 2011,7, 2419-2427

Selection of tetracycline inducible self-cleaving ribozymes as synthetic devices for gene regulation in yeast

A. Wittmann and B. Suess, Mol. BioSyst., 2011, 7, 2419 DOI: 10.1039/C1MB05070B

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