Design of RNA hairpin modules that predictably tune translation in yeast
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Published version
Author(s)
Weenink, Tim
van der Hilst, Jelle
McKiernan, Robert
Ellis, Thomas
Type
Journal Article
Abstract
Modular parts for tuning translation are prevalent in prokaryotic synthetic biology but lacking for eukaryotic synthetic biology. Working in Saccharomyces cerevisiae yeast, we here describe how hairpin RNA structures inserted into the 5′ untranslated region (5′UTR) of mRNAs can be used to tune expression levels by 100-fold by inhibiting translation. We determine the relationship between the calculated free energy of folding in the 5′UTR and in vivo protein abundance, and show that this enables rational design of hairpin libraries that give predicted expression outputs. Our approach is modular, working with different promoters and protein coding sequences, and outperforms promoter mutation as a way to predictably generate a library where a protein is induced to express at a range of different levels. With this new tool, computational RNA sequence design can be used to predictably fine-tune protein production for genes expressed in yeast.
Date Issued
2019-01-01
Date Acceptance
2018-10-05
Citation
Synthetic Biology, 2019, 3 (1)
ISSN
2397-7000
Publisher
Oxford University Press (OUP)
Journal / Book Title
Synthetic Biology
Volume
3
Issue
1
Copyright Statement
© The Author(s) 2018. Published by Oxford University Press.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://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 (http://creativecommons.org/licenses/by/
4.0/),
which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited
Sponsor
Engineering & Physical Science Research Council (EPSRC)
Biotechnology and Biological Sciences Research Council (BBSRC)
Grant Number
EP/J021849/1
BB/K019791/1
Publication Status
Published
Date Publish Online
2018-10-13