4.7 Article

Building an RNA-Based Toggle Switch Using Inhibitory RNA Aptamers

期刊

ACS SYNTHETIC BIOLOGY
卷 11, 期 2, 页码 562-569

出版社

AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.1c00580

关键词

RNA circuit; toggle switch; in vitro transcription; RNA aptamer; RNA polymerases; RNase

资金

  1. EPSRC [EP/P02596X/1]
  2. Royal Society University Research Fellowship
  3. Royal Academy of Engineering Chair in Emerging Technology for Engineering Biology [CiET1819\5]

向作者/读者索取更多资源

Synthetic RNA systems have advantages over protein-based networks in terms of faster response, increased specificity, and programmability. In this study, we developed an in vitro RNA toggle switch using RNA aptamers to inhibit the transcriptional activity of two different RNA polymerases. The switch was successfully controlled by adding DNA sequences. The results demonstrated that the RNA-based toggle switch has potential applications in synthetic biology.
Synthetic RNA systems offer unique advantages such as faster response, increased specificity, and programmability compared to conventional protein-based networks. Here, we demonstrate an in vitro RNA-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.

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