Cloning and expression of transgenes using linear vectors in Trypanosoma cruzi

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dc.contributor.authorlos Angeles Curto, Maria de
dc.contributor.authorLorenzi, Hernan A.
dc.contributor.authorMoraes Barros, Roberto R. [UNIFESP]
dc.contributor.authorSouza, Renata T. [UNIFESP]
dc.contributor.authorLevin, Mariano J.
dc.contributor.authorFranco da Silveira, José [UNIFESP]
dc.contributor.authorSchijman, Alejandro G.
dc.contributor.institutionInst Invest Ingn Genet & Biol Mol INGEBI CONICET
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.date.accessioned2016-01-24T14:37:22Z
dc.date.available2016-01-24T14:37:22Z
dc.date.issued2014-06-01
dc.description.abstractThe identification of new targets for vaccine and drug development for the treatment of Chagas' disease is dependent on deepening our understanding of the parasite genome. Vectors for genetic manipulation in Trypanosoma cruzi basically include those that remain as circular episomes and those that integrate into the parasite's genome. Artificial chromosomes are alternative vectors to overcome problematic transgene expression often occurring with conventional vectors in this parasite. We have constructed a series of vectors named pTACs (Trypanosome Artificial Chromosomes), all of them carrying telomeric and subtelomeric sequences and genes conferring resistance to different selection drugs. in addition, one pTAC harbours a modified GFP gene (pTAC-gfp), and another one carries the ornithine decarboxilase gene from Crithidia fasciculata (pTAC-odc). We have encountered artificial chromosomes generated from pTACs in transformed T. cruzi epimastigotes for every version of the designed vectors. These extragenomic elements, in approximately 6-8 copies per cell, remained as linear episomes, contained telomeres and persisted after 150 and 60 generations with or without selection drugs, respectively. the linear molecules remained stable through the different T. cruzi developmental forms. Furthermore, derived artificial chromosomes from pTAC-odc could complement the auxotrophy of T. cruzi for polyamines. Our results show that pTACs constitute useful tools for reverse functional genetics in T. cruzi that will contribute to a better understanding of T. cruzi biology. (C) 2014 Published by Elsevier B.V. on behalf of Australian Society for Parasitology Inc.en
dc.description.affiliationInst Invest Ingn Genet & Biol Mol INGEBI CONICET, Lab Biol Mol Enfermedad Chagas, Buenos Aires, DF, Argentina
dc.description.affiliationUniversidade Federal de São Paulo, Dept Microbiol Imunol & Parasitol, Escola Paulista Med, São Paulo, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Dept Microbiol Imunol & Parasitol, Escola Paulista Med, São Paulo, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipWorld Health Organization
dc.description.sponsorshipAgencia Nacional de Investigaciones Cientificas y Tecnologicas (Argentina)
dc.description.sponsorshipConsejo Nacional de Investigaciones Cientificas y Tecnologicas (CONICET,Argentina)
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipUnited Nations University-Biotechnology for Latin America and the Caribbean
dc.format.extent447-456
dc.identifierhttp://dx.doi.org/10.1016/j.ijpara.2014.03.009
dc.identifier.citationInternational Journal for Parasitology. Oxford: Elsevier B.V., v. 44, n. 7, p. 447-456, 2014.
dc.identifier.doi10.1016/j.ijpara.2014.03.009
dc.identifier.issn0020-7519
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/37827
dc.identifier.wosWOS:000338599600004
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofInternational Journal for Parasitology
dc.rightsinfo:eu-repo/semantics/restrictedAccess
dc.rights.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.subjectTiypanosoma cruzien
dc.subjectLinear vectorsen
dc.subjectArtificial chromosomesen
dc.subjectGFPen
dc.subjectStable transfectionen
dc.subjectFunctional genetic complementationen
dc.subjectOrnithine decarboxylaseen
dc.titleCloning and expression of transgenes using linear vectors in Trypanosoma cruzien
dc.typeinfo:eu-repo/semantics/article
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