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Advances in time course extracellular production of human pre-miR-29b from Rhodovulum sulfidophilum

2016-04Journal article Restricted access
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dc.contributor.authorPereira, Patrícia
dc.contributor.authorPedro, Augusto
dc.contributor.authorTomás, Joana
dc.contributor.authorBaptista, Cláudio
dc.contributor.authorQueiroz, João
dc.contributor.authorFigueiras, Ana
dc.contributor.authorSousa, Fani
dc.date.accessioned2019-12-20T12:04:31Z
dc.date.available2019-12-20T12:04:31Z
dc.date.issued2016-04
dc.description.abstractThe present study reports the successful production of human pre-miR-29b both intra- and extracellularly in the marine phototrophic bacterium Rhodovulum sulfidophilum using recombinant RNA technology. In a first stage, the optimal transformation conditions (0.025 μg of plasmid DNA, with a heat-shock of 2 min at 35 °C) were established, in order to transfer the pre-miR-29b encoding plasmid to R. sulfidophilum host. Furthermore, the extracellular recovery of this RNA product from the culture medium was greatly improved, achieving quantities that are compatible with the majority of applications, namely for in vitro or in vivo studies. Using this system, the extracellular human pre-miR-29b concentration was approximately 182 μg/L, after 40 h of bacterial growth, and the total intracellular pre-miR-29b was of about 358 μg/L, at 32 h. At the end of the fermentation, it was verified that almost 87 % of cells were viable, indicating that cell lysis is minimized and that the extracellular medium is not highly contaminated with the host intracellular ribonucleases (RNases) and endotoxins, which is a critical parameter to guarantee the microRNA (miRNA) integrity. These findings demonstrate that pre-miRNAs can be produced by recombinant RNA technology, offering novel clues for the production of natural pre-miRNA agents for functional studies and RNA interference (RNAi)-based therapeutics.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.1007/s00253-016-7350-xpt_PT
dc.identifier.urihttp://hdl.handle.net/10400.6/8000
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.subjectCulture Mediapt_PT
dc.subjectHumanspt_PT
dc.subjectMicroRNAspt_PT
dc.subjectPlasmidspt_PT
dc.subjectRhodovulumpt_PT
dc.subjectGene Expressionpt_PT
dc.titleAdvances in time course extracellular production of human pre-miR-29b from Rhodovulum sulfidophilumpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/COMPETE/EXPL%2FBBB-BIO%2F1056%2F2012/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876/PEst-OE%2FSAU%2FUI0709%2F2014/PT
oaire.citation.endPage3734pt_PT
oaire.citation.issue8pt_PT
oaire.citation.startPage3723pt_PT
oaire.citation.titleApplied Microbiology and Biotechnologypt_PT
oaire.citation.volume100pt_PT
oaire.fundingStreamCOMPETE
oaire.fundingStream5876
person.familyNamePereira
person.familyNameQuaresma Henriques Pedro
person.familyNameMelfe Tomás
person.familyNameMaia Baptista
person.familyNameQueiroz
person.familyNameFigueiras
person.familyNameSousa
person.givenNamePatrícia
person.givenNameAugusto
person.givenNameJoana Filipa
person.givenNameCláudio Jorge
person.givenNameJoão
person.givenNameAna
person.givenNameFani
person.identifierhttps://scholar.google.com/citations?user=oer44UoAAAAJ&hl=pt-PT&oi=ao
person.identifierhttps://scholar.google.pt/citations?user=hFKOd_QAAAAJ&hl=pt-PT&oi=ao
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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
rcaap.embargofctCopyright cedido à editora no momento da publicação.pt_PT
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT
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