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Genome Engineering of the Fast-Growing Mycoplasma feriruminatoris toward a Live Vaccine Chassis

hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorTALENTON, Vincent
hal.structure.identifierUniversité de Sherbrooke [UdeS]
dc.contributor.authorBABY, Vincent
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorGOURGUES, Geraldine
hal.structure.identifierBiodiversité, Gènes & Communautés [BioGeCo]
dc.contributor.authorMOUDEN, Charlotte
hal.structure.identifierPlateforme Protéome [Bordeaux]
dc.contributor.authorCLAVEROL, Stephane
hal.structure.identifierJ. Craig Venter Institute
dc.contributor.authorVASHEE, Sanjay
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorBLANCHARD, Alain
hal.structure.identifierVetsuisse Faculty
dc.contributor.authorLABROUSSAA, Fabien
hal.structure.identifierVetsuisse Faculty
dc.contributor.authorJORES, Joerg
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorARFI, Yonathan
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorSIRAND-PUGNET, Pascal
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorLARTIGUE, Carole
dc.date.issued2022-05-20
dc.identifier.issn2161-5063
dc.description.abstractEnDevelopment of a new generation of vaccines is a key challenge for the control of infectious diseases affecting both humans and animals. Synthetic biology methods offer new ways to engineer bacterial chassis that can be used as vectors to present heterologous antigens and train the immune system against pathogens. Here, we describe the construction of a bacterial chassis based on the fast-growing Mycoplasma feriruminatoris, and the first steps toward its application as a live vaccine against contagious caprine pleuropneumonia (CCPP). To do so, the M. feriruminatoris genome was cloned in yeast, modified by iterative cycles of Cas9-mediated deletion of loci encoding virulence factors, and transplanted back in Mycoplasma capricolum subsp. capricolum recipient cells to produce the designed M. feriruminatoris chassis. Deleted genes encoded the glycerol transport and metabolism systems GtsABCD and GlpOKF and the Mycoplasma Ig binding protein-Mycoplasma Ig protease (MIB-MIP) immunoglobulin cleavage system. Phenotypic assays of the M. feriruminatoris chassis confirmed the corresponding loss of H2O2 production and IgG cleavage activities, while growth remained unaltered. The resulting mycoplasma chassis was further evaluated as a platform for the expression of heterologous surface proteins. A genome locus encoding an inactivated MIB-MIP system from the CCPP-causative agent Mycoplasma capricolum subsp. capripneumoniae was grafted in replacement of its homolog at the original locus in the chassis genome. Both heterologous proteins were detected in the resulting strain using proteomics, confirming their expression. This study demonstrates that advanced genome engineering methods are henceforth available for the fast-growing M. feriruminatoris, facilitating the development of novel vaccines, in particular against major mycoplasma diseases.
dc.language.isoen
dc.publisherAmerican Chemical Society
dc.subjectpathologie animale
dc.subjectcaprin
dc.subjectBactérie pathogène
dc.subjectMycoplasme
dc.subjectMollicutes
dc.subject.enbacterial chassis
dc.subject.ensynthetic biology
dc.subject.envaccine
dc.subject.encontagious caprine pleuropneumonia
dc.subject.enMycoplasma feriruminatoris
dc.subject.engenome
dc.subject.entransplantation
dc.subject.enCRISPR-Cas9
dc.subject.engenome engineering
dc.subject.enCReasPy-cloning
dc.title.enGenome Engineering of the Fast-Growing Mycoplasma feriruminatoris toward a Live Vaccine Chassis
dc.typeArticle de revue
dc.identifier.doi10.1021/acssynbio.2c00062
dc.subject.halSciences du Vivant [q-bio]
dc.subject.halSciences du Vivant [q-bio]/Biologie animale/Médecine vétérinaire et santé animal
bordeaux.journalACS Synthetic Biology
bordeaux.page1919-1930
bordeaux.volume11
bordeaux.issue5
bordeaux.peerReviewedoui
hal.identifierhal-03675959
hal.version1
hal.popularnon
hal.audienceInternationale
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-03675959v1
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