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Sustained gene expression in the retina by improved episomal vectors

2014-10Journal article Open access
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dc.contributor.authorCalado, Sofia
dc.contributor.authorOliveira, Ana
dc.contributor.authorMachado, Susana
dc.contributor.authorHaase, Rudolf
dc.contributor.authorSilva, Gabriela
dc.date.accessioned2018-12-07T14:58:06Z
dc.date.available2018-12-07T14:58:06Z
dc.date.issued2014-10
dc.description.abstractGene and cellular therapies are nowadays part of therapeutic strategies for the treatment of diverse pathologies. The drawbacks associated with gene therapy-low levels of transgene expression, vector loss during mitosis, and gene silencing-need to be addressed. The pEPI-1 and pEPito family of vectors was developed to overcome these limitations. It contains a scaffold/matrix attachment region, which anchors its replication to cell division in eukaryotic cells while in an extrachromosomal state and is less prone to silencing, due to a lower number of CpG motifs. Recent success showed that ocular gene therapy is an important tool for the treatment of several diseases, pending the overcome of the aforementioned limitations. To achieve sustained gene delivery in the retina, we evaluated several vectors based on pEPito and pEPI-1 for their ability to sustain transgene expression in retinal cells. These vectors stably transfected and replicated in retinal pigment epithelial (RPE) cells. Expression levels were promoter dependent with constitutive promoters cytomegalovirus immediate early promoter (CMV) and human CMV enhancer/human elongation factor 1 alpha promoter yielding the highest levels of transgene expression compared with the retina-specific RPE65 promoter. When injected in C57Bl6 mice, transgene expression was sustained for at least 32 days. Furthermore, the retina-specific RPE65 promoter showed higher efficiency in vivo compared to in vitro. In this study, we demonstrate that by combining tissue-specific promoters with a mitotic stable system, less susceptible to epigenetic silencing such as pEPito-based plasmids, we can achieve prolonged gene expression and a sustained therapeutic effect.
dc.description.sponsorshipEuropean Union PIRG-GA-2009-249314
dc.identifier.doihttps://doi.org/10.1089/ten.tea.2013.0672
dc.identifier.issn1937-3341
dc.identifier.issn1937-335X
dc.identifier.urihttp://hdl.handle.net/10400.1/11858
dc.language.isoeng
dc.peerreviewedyes
dc.publisherMary Ann Liebert
dc.relationIncentive
dc.relationDEVELOPMENT OF NON-VIRAL VECTORS FOR GENE THERAPY FOR PATHOLOGIES OF THE RETINA
dc.subjectScaffold matrix attachment Rregion
dc.subjectEpithelial cell line
dc.subjectPlasmid Dna
dc.subjectCpg free
dc.subjectIn vivo
dc.subjectTherapy
dc.subjectDelivery
dc.subjectS/Mars
dc.titleSustained gene expression in the retina by improved episomal vectors
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleIncentive
oaire.awardTitleDEVELOPMENT OF NON-VIRAL VECTORS FOR GENE THERAPY FOR PATHOLOGIES OF THE RETINA
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/Incentivo%2FEQB%2FLA0023%2F2013/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/OE/SFRH%2FBD%2F76873%2F2011/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//SFRH%2FBD%2F70318%2F2010/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FSAU-BEB%2F098475%2F2008/PT
oaire.citation.endPage2698
oaire.citation.issue19-20
oaire.citation.startPage2692
oaire.citation.titleTissue Engineering Part A
oaire.citation.volume20
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStreamOE
oaire.fundingStream3599-PPCDT
person.familyNameCalado
person.familyNameOliveira
person.familyNameMachado
person.familyNameSilva
person.givenNameSofia
person.givenNameAna
person.givenNameSusana
person.givenNameGabriela
person.identifier.ciencia-id7C1A-91BA-B6A0
person.identifier.ciencia-idCA17-42CB-F11B
person.identifier.ciencia-idDC1E-F164-FDD9
person.identifier.orcid0000-0001-5509-4145
person.identifier.orcid0000-0003-3162-250X
person.identifier.orcid0000-0002-3152-1701
person.identifier.orcid0000-0001-5946-1918
person.identifier.ridK-2202-2016
person.identifier.ridK-4033-2012
person.identifier.ridF-4409-2012
person.identifier.scopus-author-id56426215600
person.identifier.scopus-author-id55989431200
person.identifier.scopus-author-id56201414700
person.identifier.scopus-author-id7101647767
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccess
rcaap.typearticle
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