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Conditional control of Mammalian gene expression by tetracycline-dependent hammerhead ribozymes.

Beilstein, Kim and Wittmann, Alexander and Grez, Manuel and Suess, Beatrix (2015):
Conditional control of Mammalian gene expression by tetracycline-dependent hammerhead ribozymes.
4, In: ACS synthetic biology, (5), pp. 526-34, ISSN 2161-5063, [Article]

Abstract

Robust synthetic devices are requisite for the construction of synthetic genetic circuits and important scientific and technological tools to control cellular processes. We developed tetracycline-dependent ribozymes, which can switch on gene expression up to 8.7-fold upon addition of tetracycline. A tetracycline aptamer was grafted onto the hammerhead ribozyme in such a way that ligand binding to the aptamers destroys a loop-loop interaction within the ribozyme thereby inhibiting ribozyme cleavage and allowing gene expression. The advantage of the presented regulatory system is its independence of any regulatory proteins. The stable integration of the ribozyme into the genome of HeLa cells indicates a low background activity in the absence of ligand. Furthermore, the ligand concentration required to robustly flip the switch does not affect cell viability and therefore allows a long-term application of the system. These properties turn the tetracycline-dependent ribozymes into a very promising tool for conditional gene expression in mammalian cells.

Item Type: Article
Erschienen: 2015
Creators: Beilstein, Kim and Wittmann, Alexander and Grez, Manuel and Suess, Beatrix
Title: Conditional control of Mammalian gene expression by tetracycline-dependent hammerhead ribozymes.
Language: English
Abstract:

Robust synthetic devices are requisite for the construction of synthetic genetic circuits and important scientific and technological tools to control cellular processes. We developed tetracycline-dependent ribozymes, which can switch on gene expression up to 8.7-fold upon addition of tetracycline. A tetracycline aptamer was grafted onto the hammerhead ribozyme in such a way that ligand binding to the aptamers destroys a loop-loop interaction within the ribozyme thereby inhibiting ribozyme cleavage and allowing gene expression. The advantage of the presented regulatory system is its independence of any regulatory proteins. The stable integration of the ribozyme into the genome of HeLa cells indicates a low background activity in the absence of ligand. Furthermore, the ligand concentration required to robustly flip the switch does not affect cell viability and therefore allows a long-term application of the system. These properties turn the tetracycline-dependent ribozymes into a very promising tool for conditional gene expression in mammalian cells.

Journal or Publication Title: ACS synthetic biology
Volume: 4
Number: 5
Divisions: 10 Department of Biology
10 Department of Biology > Synthetic Genetic Circuits
Date Deposited: 30 Jun 2015 08:16
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