Conditional Control of Mammalian Gene Expression by Tetracycline-Dependent Hammerhead Ribozymes
Conditional Control of Mammalian Gene Expression by Tetracycline-Dependent Hammerhead Ribozymes
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DOI：
10.1021/sb500270h
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发表时间：
2015-05-01
影响因子：
4.7
通讯作者：
Suess, Beatrix
中科院分区：
文献类型：
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作者：
Beilstein, Kim;Wittmann, Alexander;Suess, Beatrix
强大的合成设备是构建合成基因电路和控制细胞过程的重要科学和技术工具的必要条件。我们开发了四环素依赖性核酶，它可以在添加四环素后打开基因表达高达8.7倍。将四环素适体以这样的方式接枝到锤头状核酶上，使得配体与适体的结合破坏核酶内的环-环相互作用，从而抑制核酶切割并允许基因表达。所提出的调节系统的优点是其独立于任何调节蛋白。核酶稳定整合到HeLa细胞基因组中表明在不存在配体的情况下具有低背景活性。此外，稳健地翻转开关所需的配体浓度不影响细胞活力，因此允许系统的长期应用。这些特性使四环素依赖性核酶成为哺乳动物细胞中条件基因表达的非常有前途的工具。
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.