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dc.contributor.author
Oesterle, Sabine
dc.contributor.author
Gerngross, Daniel
dc.contributor.author
Schmitt, Steven
dc.contributor.author
Roberts, Tania M.
dc.contributor.author
Panke, Sven
dc.date.accessioned
2017-09-26T14:43:00Z
dc.date.available
2017-09-26T13:49:58Z
dc.date.available
2017-09-26T14:43:00Z
dc.date.issued
2017
dc.identifier.issn
2045-2322
dc.identifier.other
10.1038/s41598-017-12395-3
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/187347
dc.identifier.doi
10.3929/ethz-b-000187347
dc.description.abstract
Multiplexed gene expression optimization via modulation of gene translation efficiency through ribosome binding site (RBS) engineering is a valuable approach for optimizing artificial properties in bacteria, ranging from genetic circuits to production pathways. Established algorithms design smart RBS-libraries based on a single partially-degenerate sequence that efficiently samples the entire space of translation initiation rates. However, the sequence space that is accessible when integrating the library by CRISPR/Cas9-based genome editing is severely restricted by DNA mismatch repair (MMR) systems. MMR efficiency depends on the type and length of the mismatch and thus effectively removes potential library members from the pool. Rather than working in MMR-deficient strains, which accumulate off-target mutations, or depending on temporary MMR inactivation, which requires additional steps, we eliminate this limitation by developing a pre-selection rule of genome-library-optimized-sequences (GLOS) that enables introducing large functional diversity into MMR-proficient strains with sequences that are no longer subject to MMR-processing. We implement several GLOS-libraries in Escherichia coli and show that GLOS-libraries indeed retain diversity during genome editing and that such libraries can be used in complex genome editing operations such as concomitant deletions. We argue that this approach allows for stable and efficient fine tuning of chromosomal functions with minimal effort.
en_US
dc.format
application/pdf
dc.language.iso
en
en_US
dc.publisher
Nature Publishing Group
en_US
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.title
Efficient engineering of chromosomal ribosome binding site libraries in mismatch repair proficient Escherichia coli
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 4.0 International
dc.date.published
2017-09-26
ethz.journal.title
Scientific Reports
ethz.journal.volume
7
en_US
ethz.journal.issue
1
en_US
ethz.journal.abbreviated
Sci Rep
ethz.pages.start
12327
en_US
ethz.size
10 p.
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.identifier.wos
ethz.identifier.scopus
ethz.publication.place
London
en_US
ethz.publication.status
published
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02060 - Dep. Biosysteme / Dep. of Biosystems Science and Eng.::03602 - Panke, Sven / Panke, Sven
en_US
ethz.leitzahl.certified
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02060 - Dep. Biosysteme / Dep. of Biosystems Science and Eng.::03602 - Panke, Sven / Panke, Sven
en_US
ethz.date.deposited
2017-09-26T13:49:58Z
ethz.source
FORM
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2017-09-26T14:43:03Z
ethz.rosetta.lastUpdated
2021-02-14T18:49:16Z
ethz.rosetta.versionExported
true
ethz.COinS
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