Toehold switches: de-novo-designed regulators of gene expression.

Toehold switches: de-novo-designed regulators of gene expression.
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DOI:
10.1016/j.cell.2014.10.002
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发表时间:
2014-11-06
期刊:
影响因子:
64.5
通讯作者:
Yin P
Yin P
中科院分区:
生物学1区
文献类型:
--
作者:
Green AA;Silver PA;Collins JJ;Yin P

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在活细胞中构建合成网络的努力一直受到提供宽动态范围和低串扰的调节组件数量的限制。在这里,我们报告了一类新的由从头设计的原核生物核转录调控因子,称为Toehold开关,它可以激活基因表达,以响应具有任意序列的同源RNA。Toehold开关提供高水平的正交性,并可进行正向设计,以提供高于400的平均动态范围。我们证明开关可以整合到基因组中来调节内源基因,并将它们用作响应内源RNA的传感器。我们利用脚趾开关的正交性,独立地调控12个基因,并构建了一个评估4输入和逻辑的遗传电路。Toehold开关具有宽的动态范围、正交性和可编程性,代表着一个多功能和强大的翻译调节平台,在分子生物学、合成生物学和生物技术中提供了不同的应用。
Efforts to construct synthetic networks in living cells have been hindered by the limited number of regulatory components that provide wide dynamic range and low crosstalk. Here, we report a new class of de-novo-designed prokaryotic riboregulators called toehold switches that activate gene expression in response to cognate RNAs with arbitrary sequences. Toehold switches provide a high level of orthogonality and can be forward-engineered to provide average dynamic range above 400. We show that switches can be integrated into the genome to regulate endogenous genes and use them as sensors that respond to endogenous RNAs. We exploit the orthogonality of toehold switches to regulate 12 genes independently and to construct a genetic circuit that evaluates 4-input AND logic. Toehold switches, with their wide dynamic range, orthogonality, and programmability, represent a versatile and powerful platform for regulation of translation, offering diverse applications in molecular biology, synthetic biology, and biotechnology.
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