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dc.contributor.authorAdaui, Vanessa
dc.contributor.authorKröber-Boncardo, Constanze
dc.contributor.authorBrinker, Christine
dc.contributor.authorZirpel, Henner
dc.contributor.authorSellau, Julie
dc.contributor.authorArévalo, Jorge
dc.contributor.authorDujardin, Jean Claude
dc.contributor.authorClos, Joachim
dc.date.accessioned2021-04-14T12:47:42Z
dc.date.available2021-04-14T12:47:42Z
dc.date.issued2020-10-01
dc.identifier.doi10.3390/genes11101159
dc.identifier.urihttp://hdl.handle.net/10757/655510
dc.description.abstractThe protozoan parasite Leishmania (Viannia) braziliensis (L. braziliensis) is the main cause of human tegumentary leishmaniasis in the New World, a disease affecting the skin and/or mucosal tissues. Despite its importance, the study of the unique biology of L. braziliensis through reverse genetics analyses has so far lagged behind in comparison with Old World Leishmania spp. In this study, we successfully applied a cloning-free, PCR-based CRISPR–Cas9 technology in L. braziliensis that was previously developed for Old World Leishmania major and New World L. mexicana species. As proof of principle, we demonstrate the targeted replacement of a transgene (eGFP) and two L. braziliensis single-copy genes (HSP23 and HSP100). We obtained homozygous Cas9-free HSP23-and HSP100-null mutants in L. braziliensis that matched the phenotypes reported previously for the respective L. donovani null mutants. The function of HSP23 is indeed conserved throughout the Trypanosomatida as L. major HSP23 null mutants could be complemented phenotypically with transgenes from a range of trypanosomatids. In summary, the feasibility of genetic manipulation of L. braziliensis by CRISPR–Cas9-mediated gene editing sets the stage for testing the role of specific genes in that parasite’s biology, including functional studies of virulence factors in relevant animal models to reveal novel therapeutic targets to combat American tegumentary leishmaniasis.en_US
dc.description.sponsorshipAlexander von Humboldt-Stiftungen_US
dc.formatapplication/pdfen_US
dc.language.isoengen_US
dc.publisherMDPI AGen_US
dc.relation.urlhttps://www.mdpi.com/2073-4425/11/10/1159en_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.rightsAttribution-NonCommercial-ShareAlike 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.sourceUniversidad Peruana de Ciencias Aplicadas (UPC)es_PE
dc.sourceRepositorio Academico - UPCes_PE
dc.subjectCRISPR–Cas9en_US
dc.subjectGene targetingen_US
dc.subjectHeat shock proteinsen_US
dc.subjectLeishmania braziliensisen_US
dc.subjectPhenotypingen_US
dc.subjectReverse geneticsen_US
dc.subjectAnimal cellen_US
dc.subjectArticleen_US
dc.subjectControlled studyen_US
dc.subjectCRISPR-CAS9 systemen_US
dc.titleApplication of crispr/cas9-based reverse genetics in leishmania braziliensis: Conserved roles for hsp100 and hsp23en_US
dc.typeinfo:eu-repo/semantics/articleen_US
dc.identifier.eissn20734425
dc.identifier.journalGenesen_US
dc.description.peerreviewRevisión por pareses_PE
dc.identifier.eid2-s2.0-85091874138
dc.identifier.scopusidSCOPUS_ID:85091874138
dc.source.journaltitleGenes
dc.source.volume11
dc.source.issue10
dc.source.beginpage1
dc.source.endpage24
refterms.dateFOA2021-04-14T12:47:43Z
dc.identifier.isni0000 0001 2196 144X


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