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In-Yeast Engineering of a Bacterial Genome Using CRISPR/Cas9

hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorTSARMPOPOULOS, Iason
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorGOURGUES, Geraldine
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorBLANCHARD, Alain
hal.structure.identifierJ. Craig Venter Institute [La Jolla, USA] [JCVI]
dc.contributor.authorVASHEE, Sanjay
hal.structure.identifierInternational Livestock Research Institute [CGIAR, Nairobi] [ILRI]
dc.contributor.authorJORES, Joerg
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorLARTIGUE, Carole
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorSIRAND-PUGNET, Pascal
dc.date.issued2016
dc.identifier.issn2161-5063
dc.description.abstractEnOne remarkable achievement in synthetic biology was the reconstruction of mycoplasma genomes and their cloning in yeast where they can be modified using available genetic tools. Recently, CRISPR/Cas9 editing tools were developed for yeast mutagenesis. Here, we report their adaptation for the engineering of bacterial genomes cloned in yeast. A seamless deletion of the mycoplasma glycerol-3-phosphate oxidase-encoding gene (glpO) was achieved without selection in one step, using 90 nt paired oligonucleotides as templates to drive recombination. Screening of the resulting clones revealed that more than 20% contained the desired deletion. After manipulation, the overall integrity of the cloned mycoplasma genome was verified by multiplex PCR and PFGE. Finally, the edited genome was back-transplanted into a mycoplasma recipient cell. In accordance with the deletion of glpO, the mutant mycoplasma was affected in the production of H2O2. This work paves the way to high-throughput manipulation of natural or synthetic genomes in yeast.
dc.language.isoen
dc.publisherAmerican Chemical Society
dc.subjectCRISPR/Cas9
dc.subjectSaccharomyces cerevisiae
dc.subjectgenome transplantation
dc.subject.enMycoplasma
dc.subject.engenome engineering
dc.subject.enseamless gene deletion
dc.title.enIn-Yeast Engineering of a Bacterial Genome Using CRISPR/Cas9
dc.typeArticle de revue
dc.identifier.doi10.1021/acssynbio.5b00196
dc.subject.halSciences du Vivant [q-bio]/Microbiologie et Parasitologie
dc.subject.halSciences du Vivant [q-bio]/Biologie végétale
dc.subject.halSciences du Vivant [q-bio]/Biologie végétale/Phytopathologie et phytopharmacie
bordeaux.journalACS Synthetic Biology
bordeaux.page104-109
bordeaux.volume5
bordeaux.issue1
bordeaux.peerReviewedoui
hal.identifierhal-02640111
hal.version1
hal.popularnon
hal.audienceInternationale
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-02640111v1
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