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dc.contributor.CRUESPUNIVERSIDADE DE ESTADUAL DE CAMPINASpt_BR
dc.typeArtigopt_BR
dc.titleProduction Of Dna Minicircles Less Than 250 Base Pairs Through A Novel Concentrated Dna Circularization Assay Enabling Minicircle Design With Nf-κb Inhibition Activityen
dc.contributor.authorThibault T.pt_BR
dc.contributor.authorDegrouard J.pt_BR
dc.contributor.authorBaril P.pt_BR
dc.contributor.authorPichon C.pt_BR
dc.contributor.authorMidoux P.pt_BR
dc.contributor.authorMalinge J.-M.pt_BR
unicamp.authorUniversidade Estadual de Campinas, Hemocentro, Campinas-São Paulo, Brazilpt_BR
unicamp.author.externalCentre de Biophysique Moléculaire, CNRS UPR 4301, University of Orléans and Inserm, Orléans Cedex 02, Francept_BR
unicamp.author.externalLaboratoire de Physique des Solides, Université Paris Sud, CNRS UMR 8502, Orsay Cedex, Francept_BR
dc.description.abstractDouble-stranded DNA minicircles of less than 1000 bp in length have great interest in both fundamental research and therapeutic applications. Although minicircles have shown promising activity in gene therapy thanks to their good biostability and better intracellular trafficking, minicircles down to 250 bp in size have not yet been investigated from the test tube to the cell for lack of an efficient production method. Herein, we report a novel versatile plasmidfree method for the production of DNA minicircles comprising fewer than 250 bp. We designed a linear nicked DNA double-stranded oligonucleotide bluntended substrate for efficient minicircle production in a ligase-mediated and bending protein-assisted circularization reaction at high DNA concentration of 2M. This one pot multi-step reaction based-method yields hundreds of micrograms of minicircle with sequences of any base composition and position and containing or not a variety of site-specifically chemical modifications or physiological supercoiling. Biochemical and cellular studies were then conducted to design a 95 bp minicircle capable of binding in vitro two NF-κB transcription factors per minicircle and to efficiently inhibiting NF-κB-dependent transcriptional activity in human cells. Therefore, our production method could pave the way for the design of minicircles as new decoy nucleic acids. © The Author(s) 2016.en
dc.relation.ispartofNucleic Acids Researchpt_BR
dc.publisherOxford University Presspt_BR
dc.date.issued2017pt_BR
dc.identifier.citationNucleic Acids Research. Oxford University Press, v. 45, n. 5, p. , 2017.pt_BR
dc.language.isoEnglishpt_BR
dc.description.volume45pt_BR
dc.description.issuenumber5pt_BR
dc.rightsabertopt_BR
dc.sourceScopuspt_BR
dc.identifier.issn0305-1048pt_BR
dc.identifier.doi10.1093/nar/gkw1034pt_BR
dc.identifier.urlhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85018301374&doi=10.1093%2fnar%2fgkw1034&partnerID=40&md5=c3f524d39795244707d98d62e726a0bapt_BR
dc.date.available2017-08-17T19:12:58Z-
dc.date.accessioned2017-08-17T19:12:58Z-
dc.description.provenanceMade available in DSpace on 2017-08-17T19:12:58Z (GMT). No. of bitstreams: 1 2-s2.0-85018301374.pdf: 4109702 bytes, checksum: 79719901ab9495ee79a3c9f6196aec4d (MD5) Previous issue date: 2017en
dc.identifier.urihttp://repositorio.unicamp.br/jspui/handle/REPOSIP/323239-
dc.identifier.idScopus2-s2.0-85018301374pt_BR
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