Building an RNA-based toggle switch using inhibitory RNA aptamers

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Title: Building an RNA-based toggle switch using inhibitory RNA aptamers
Authors: Climent-Catala, A
Ouldridge, TE
Stan, G-BV
Bae, W
Item Type: Journal Article
Abstract: 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.
Issue Date: 18-Feb-2022
Date of Acceptance: 1-Feb-2022
URI: http://hdl.handle.net/10044/1/95042
DOI: 10.1021/acssynbio.1c00580
ISSN: 2161-5063
Publisher: American Chemical Society
Start Page: 562
End Page: 569
Journal / Book Title: ACS Synthetic Biology
Volume: 11
Issue: 2
Copyright Statement: © 2022 American Chemical Society
Sponsor/Funder: The Royal Society
Engineering & Physical Science Research Council (EPSRC)
Funder's Grant Number: UF150067
EP/P02596X/1
Keywords: RNA aptamer
RNA circuit
RNA polymerases
RNase
in vitro transcription
toggle switch
RNA aptamer
RNA circuit
RNA polymerases
RNase
in vitro transcription
toggle switch
0304 Medicinal and Biomolecular Chemistry
0601 Biochemistry and Cell Biology
0903 Biomedical Engineering
Publication Status: Published
Conference Place: United States
Embargo Date: Embargoed for 12 months after publication date
Online Publication Date: 2022-02-08
Appears in Collections:Bioengineering