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dc.contributor.authorWendisch, Volker F.
dc.contributor.authorCleto, Sara
dc.contributor.authorJensen, Jaide
dc.contributor.authorLu, Timothy K
dc.date.accessioned2018-02-20T14:53:38Z
dc.date.available2018-02-20T14:53:38Z
dc.date.issued2016-02
dc.date.submitted2015-11
dc.identifier.issn2161-5063
dc.identifier.urihttp://hdl.handle.net/1721.1/113825
dc.description.abstractCorynebacterium glutamicum is an important organism for the industrial production of amino acids. Metabolic pathways in this organism are usually engineered by conventional methods such as homologous recombination, which depends on rare double-crossover events. To facilitate the mapping of gene expression levels to metabolic outputs, we applied CRISPR interference (CRISPRi) technology using deactivated Cas9 (dCas9) to repress genes in C. glutamicum. We then determined the effects of target repression on amino acid titers. Single-guide RNAs directing dCas9 to specific targets reduced expression of pgi and pck up to 98%, and of pyk up to 97%, resulting in titer enhancement ratios of l-lysine and l-glutamate production comparable to levels achieved by gene deletion. This approach for C. glutamicum metabolic engineering, which only requires 3 days, indicates that CRISPRi can be used for quick and efficient metabolic pathway remodeling without the need for gene deletions or mutations and subsequent selection. Keywords: amino acid; C. glutamicum; CRISPRi; metabolic engineering; sgRNA/dCas9en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (DP2 OD008435)en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (P50 GM098792)en_US
dc.description.sponsorshipUnited States. Office of Naval Research (N00014-13-1-0424)en_US
dc.description.sponsorshipNational Science Foundation (U.S.) (MCB1350625)en_US
dc.description.sponsorshipWuxi NewWay Biotech Ltd. (Jiangsu Province International Collaboration Grant BZ2014014)en_US
dc.language.isoen_US
dc.publisherAmerican Chemical Societyen_US
dc.relation.isversionofhttp://dx.doi.org/10.1021/acssynbio.5b00216en_US
dc.rightsArticle is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.en_US
dc.sourceACSen_US
dc.titleCorynebacterium glutamicum Metabolic Engineering with CRISPR Interference (CRISPRi)en_US
dc.typeArticleen_US
dc.identifier.citationCleto, Sara, Jaide VK Jensen, Volker F. Wendisch, and Timothy K. Lu. “Corynebacterium glutamicumMetabolic Engineering with CRISPR Interference (CRISPRi).” ACS Synthetic Biology 5, no. 5 (May 20, 2016): 375–385.en_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biological Engineeringen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Electrical Engineering and Computer Scienceen_US
dc.contributor.departmentMassachusetts Institute of Technology. Synthetic Biology Centeren_US
dc.contributor.mitauthorCleto, Sara
dc.contributor.mitauthorJensen, Jaide
dc.contributor.mitauthorLu, Timothy K
dc.relation.journalACS Synthetic Biologyen_US
dc.eprint.versionAuthor's final manuscripten_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dspace.orderedauthorsCleto, Sara; Jensen, Jaide VK; Wendisch, Volker F.; Lu, Timothy K.en_US
dspace.embargo.termsNen_US
dc.identifier.orcidhttps://orcid.org/0000-0002-9999-6690
mit.licensePUBLISHER_POLICYen_US


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