Building an RNA-based Toggle Switch using Inhibitory RNA Aptamers
Building an RNA-based Toggle Switch using Inhibitory RNA Aptamers
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使用抑制性 RNA 适体构建基于 RNA 的拨动开关
DOI：
10.1101/2021.11.16.468831
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
Climent-Catala A
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文献类型：
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作者：
Climent-Catala A
与传统的基于蛋白质的网络相比，合成RNA系统具有独特的优势，如更快的响应速度、更高的特异性和可编程性。在这里，我们证明了一种基于体外RNA的触发开关，使用RNA适配子能够抑制T7或SP6 RNA聚合酶的转录活性。用花椰菜和孔雀石绿光适配子系统同时监测两种聚合酶的活性。在我们的触发开关中，T7启动子驱动SP6抑制适配子的表达，SP6启动子表达T7抑制适配子。我们证明了通过添加DNA序列来隔离RNA抑制适配子，可以切换源于相互抑制适配子的两种不同的状态。最后，我们评估了我们的基于RNA的触发开关在降解条件下，通过引入使用混合RNase的RNA的受控降解。我们的结果表明，基于RNA的触发开关可以作为核酸网络在合成生物学应用中的控制元件。
Synthetic RNA systems offer unique advantages such as faster response, increased specificity, and programmability compared to conventional protein-based networks. Here, we demonstrate anin vitroRNA-based toggle switch using RNA aptamers capable of inhibiting the transcriptional activity of T7 or SP6 RNA polymerases. The activities of both polymerases are monitored simultaneously by using Broccoli and malachite green light-up aptamer systems. In our toggle switch, a T7 promoter drives the expression of SP6 inhibitory aptamers, and an SP6 promoter expresses T7 inhibitory aptamers. We show that the two distinct states originating from the mutual inhibition of aptamers can be toggled by adding DNA sequences to sequester the RNA inhibitory aptamers. Finally, we assessed our RNA-based toggle switch in degrading conditions by introducing controlled degradation of RNAs using a mix of RNases. Our results demonstrate that the RNA-based toggle switch could be used as a control element for nucleic acid networks in synthetic biology applications.