Design of RNA hairpin modules that predictably tune translation in yeast

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Title: Design of RNA hairpin modules that predictably tune translation in yeast
Authors: Weenink, T
Van der Hilst, J
McKiernan, R
Ellis, T
Item 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.
Issue Date: 1-Jan-2019
Date of Acceptance: 5-Oct-2018
URI: http://hdl.handle.net/10044/1/65245
DOI: https://dx.doi.org/10.1093/synbio/ysy019
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
Sponsor/Funder: Engineering & Physical Science Research Council (EPSRC)
Biotechnology and Biological Sciences Research Council (BBSRC)
Funder's Grant Number: EP/J021849/1
BB/K019791/1
Publication Status: Published
Online Publication Date: 2018-10-13
Appears in Collections:Faculty of Engineering
Bioengineering



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