Building an RNA-based toggle switch using inhibitory RNA aptamers
File(s)acssynbio.1c00580.pdf (1.4 MB)
Published version
Author(s)
Climent-Catala, Alicia
Ouldridge, Thomas E
Stan, Guy-Bart V
Bae, Wooli
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.
Date Issued
2022-02-18
Date Acceptance
2022-02-01
Citation
ACS Synthetic Biology, 2022, 11 (2), pp.562-569
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. This work is published under a CC BY licence.
License URL
Sponsor
The Royal Society
Engineering & Physical Science Research Council (EPSRC)
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/35133150
Grant Number
UF150067
EP/P02596X/1
Subjects
RNA aptamer
RNA circuit
RNA polymerases
RNase
in vitro transcription
toggle switch
Publication Status
Published
Coverage Spatial
United States
Date Publish Online
2022-02-08