A modular RNA interference system for multiplexed gene regulation
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Published online version
Author(s)
Dwijayanti, Ari
Storch, Marko
Stan, Guy-Bart
Baldwin, Geoff S
Type
Journal Article
Abstract
The rational design and realisation of simple-to-use genetic control elements that are modular, orthogonal and robust is essential to the construction of predictable and reliable biological systems of increasing complexity. To this effect, we introduce modular Artificial RNA interference (mARi), a rational, modular and extensible design framework that enables robust, portable and multiplexed post-transcriptional regulation of gene expression in Escherichia coli. The regulatory function of mARi was characterised in a range of relevant genetic contexts, demonstrating its independence from other genetic control elements and the gene of interest, and providing new insight into the design rules of RNA based regulation in E. coli, while a range of cellular contexts also demonstrated it to be independent of growth-phase and strain type. Importantly, the extensibility and orthogonality of mARi enables the simultaneous post-transcriptional regulation of multi-gene systems as both single-gene cassettes and poly-cistronic operons. To facilitate adoption, mARi was designed to be directly integrated into the modular BASIC DNA assembly framework. We anticipate that mARi-based genetic control within an extensible DNA assembly framework will facilitate metabolic engineering, layered genetic control, and advanced genetic circuit applications.
Date Issued
2022-01-21
Date Acceptance
2021-12-22
Citation
Nucleic Acids Research, 2022, 50 (3)
ISSN
0305-1048
Publisher
Oxford University Press
Journal / Book Title
Nucleic Acids Research
Volume
50
Issue
3
Copyright Statement
© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which
permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which
permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
License URL
Sponsor
Engineering & Physical Science Research Council (EPSRC)
Royal Academy Of Engineering
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/35061908
PII: 6513569
Grant Number
EP/M002187/1
CiET1819\5
Subjects
Developmental Biology
05 Environmental Sciences
06 Biological Sciences
08 Information and Computing Sciences
Publication Status
Published online
Coverage Spatial
England
Date Publish Online
2022-01-21