A split intein T7 RNA polymerase for transcriptional AND-logic

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• dc.contributor.author Schaerli, Yolanda
• dc.contributor.author Gili, Magui
• dc.contributor.author Isalan, Mark
• dc.date.accessioned 2025-01-13T08:35:21Z
• dc.date.available 2025-01-13T08:35:21Z
• dc.date.issued 2014
• dc.description.abstract Synthetic biology has developed numerous parts for building synthetic gene circuits. However, few parts have been described for prokaryotes to integrate two signals at a promoter in an AND fashion, i.e. the promoter is only activated in the presence of both signals. Here we present a new part for this function: a split intein T7 RNA polymerase. We divide T7 RNA polymerase into two expression domains and fuse each to a split intein. Only when both domains are expressed does the split intein mediate protein trans-splicing, yielding a full-length T7 RNA polymerase that can transcribe genes via a T7 promoter. We demonstrate an AND gate with the new part: the signal-to-background ratio is very high, resulting in an almost digital signal. This has utility for more complex circuits and so we construct a band-pass filter in Escherichia coli. The split intein approach should be widely applicable for engineering artificial gene circuit parts.en
• dc.description.sponsorship Spanish Ministry of Economy and Competitiveness, Centro de Excelencia Severo Ochoa 2013–2017 [SEV-2012-0208]; Marie Curie Action FP7 [PIEF-GA-2011-298348 to Y.S.]; European Research Council [FP7-ERC-201249-ZINC-HUBS], European Commission [FP7-610730 EVOPROG]; Wellcome Trust UK New Investigator Award [WT102944 to M.I.]. Funding for open access charge: Wellcome Trust UK New Investigator Award [WT102944].en
• dc.format.mimetype application/pdf
• dc.identifier.citation Schaerli Y, Gili M, Isalan M. A split intein T7 RNA polymerase for transcriptional AND-logic. Nucleic Acids Res. 2014 Oct 29;42(19):12322-8. DOI: 10.1093/nar/gku884
• dc.identifier.doi http://dx.doi.org/10.1093/nar/gku884
• dc.identifier.issn 0305-1048
• dc.identifier.uri http://hdl.handle.net/10230/69064
• dc.language.iso eng
• dc.publisher Oxford University Press
• dc.relation.ispartof Nucleic Acids Research. 2014 Oct 29;42(19):12322-8
• dc.relation.projectID info:eu-repo/grantAgreement/EC/FP7/298348
• dc.relation.projectID info:eu-repo/grantAgreement/ES/1PE/SEV-2012-0208
• dc.relation.projectID info:eu-repo/grantAgreement/EC/FP7/610730
• dc.rights © The Author(s) 2014. 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 (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
• dc.rights.accessRights info:eu-repo/semantics/openAccess
• dc.rights.uri http://creativecommons.org/licenses/by/4.0/
• dc.subject.other Reacció en cadena de la polimerasaca
• dc.subject.other Procariotesca
• dc.subject.other Proteïnesca
• dc.title A split intein T7 RNA polymerase for transcriptional AND-logic
• dc.type info:eu-repo/semantics/article
• dc.type.version info:eu-repo/semantics/publishedVersion