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dc.contributor.authorGeisler, Katrin
dc.contributor.authorScaife, Mark A
dc.contributor.authorMordaka, Paweł M
dc.contributor.authorHolzer, Andre
dc.contributor.authorTomsett, Eleanor V
dc.contributor.authorMehrshahi, Payam
dc.contributor.authorMendoza Ochoa, Gonzalo I
dc.contributor.authorSmith, Alison G
dc.date.accessioned2021-10-30T01:12:14Z
dc.date.available2021-10-30T01:12:14Z
dc.date.issued2021-09-14
dc.identifier.citationLife (Basel, Switzerland), volume 11, issue 9
dc.identifier.issn2075-1729
dc.identifier.otherPMC8471596
dc.identifier.other34575113
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/330081
dc.description.abstract<i>Chlamydomonas reinhardtii</i> has many attractive features for use as a model organism for both fundamental studies and as a biotechnological platform. Nonetheless, despite the many molecular tools and resources that have been developed, there are challenges for its successful engineering, in particular to obtain reproducible and high levels of transgene expression. Here we describe a synthetic biology approach to screen several hundred independent transformants using standardised parts to explore different parameters that might affect transgene expression. We focused on terminators and, using a standardised workflow and quantitative outputs, tested 9 different elements representing three different size classes of native terminators to determine their ability to support high level expression of a GFP reporter gene. We found that the optimal size reflected the median size of element found in the <i>C. reinhardtii</i> genome. The behaviour of the terminator parts was similar with different promoters, in different host strains and with different transgenes. This approach is applicable to the systematic testing of other genetic elements, facilitating comparison to determine optimal transgene design.
dc.languageeng
dc.rightsAttribution 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.sourceessn: 2075-1729
dc.sourcenlmid: 101580444
dc.subjectAlgae
dc.subjectTerminator
dc.subjectHeterologous protein expression
dc.subjectModular Assembly
dc.subject3′ Utr
dc.subjectStandardized Workflow
dc.subjectQuantification Of Transgene Expression
dc.titleExploring the Impact of Terminators on Transgene Expression in <i>Chlamydomonas reinhardtii</i> with a Synthetic Biology Approach.
dc.typeArticle
dc.date.updated2021-10-30T01:12:13Z
dc.identifier.doi10.17863/CAM.77525
rioxxterms.versionofrecord10.3390/life11090964
rioxxterms.versionVoR
rioxxterms.licenseref.urihttps://creativecommons.org/licenses/by/4.0/
dc.contributor.orcidGeisler, Katrin [0000-0002-7630-1868]
dc.contributor.orcidHolzer, Andre [0000-0003-2439-6364]
dc.contributor.orcidMendoza Ochoa, Gonzalo I [0000-0002-7361-8915]
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (BB/M018180/1, BB/I007660/1, BB/L002957/1, BB/L014130/1, BB/R01860X/1)
pubs.funder-project-idBill and Melinda Gates Foundation (OPP1144)
pubs.funder-project-idSeventh Framework Programme (311956)


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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International