Please use this identifier to cite or link to this item: http://hdl.handle.net/10316/25534
DC FieldValueLanguage
dc.contributor.authorCardoso, Ana M.-
dc.contributor.authorCalejo, M. Teresa-
dc.contributor.authorMorais, Catarina M.-
dc.contributor.authorCardoso, Ana L.-
dc.contributor.authorCruz, Rita-
dc.contributor.authorZhu, Kaizheng-
dc.contributor.authorLima, M. C. P. de-
dc.contributor.authorJurado, A. S.-
dc.contributor.authorNyström, Bo-
dc.date.accessioned2014-04-10T11:04:24Z-
dc.date.available2014-04-10T11:04:24Z-
dc.date.issued2014-
dc.identifier.urihttp://hdl.handle.net/10316/25534-
dc.description.abstractGene knockdown has emerged as an important tool for cancer gene therapy as well as for viral infections and dominantly inherited genetic disorders. The generation of suitable siRNA delivery systems poses some challenges, namely, to avoid nuclease degradation, to surpass the cytoplasmic membrane, and to release the nucleic acids into the cytosol. Aiming at evaluating the ability of thermoresponsive block copolymers formed by units of N-isopropylacrylamide and of (3- acrylamidopropyl)trimethylammonium chloride to efficiently deliver siRNAs, an extensive study was performed with four different copolymers using a human fibrosarcoma cell line as cell model. The silencing ability and cytotoxicity of the generated copolymer-based siRNA delivery systems were found to be dependent on the cloud point of the polymer, which corresponds to the transition temperature at which the aggregation or precipitation of the polymer molecules becomes thermodynamically more favorable than their solubilization. In the present study, a system capable of delivering siRNAs efficiently, specifically and without presenting relevant cytotoxicity, even in the presence of serum, was developed. Confocal fluorescence experiments showed that the ability of the generated systems to silence the target gene is related to some extent to nucleic acid internalization, being also dependent on polymer/siRNA dissociation at 37 °C. Thus, a delicate balance between nucleic acid internalization and intracellular release must be met in order to reach an ideal knockdown efficiency. The special features and potential for manipulation of the N-isopropylacrylamide-based copolymers make them suitable materials for the design and synthesis of new and promising siRNA delivery systems.por
dc.description.sponsorshipThis work was supported by the Portuguese Foundation for Science and Technology and FEDER/COMPETE (research grants PTDC/QUI-BIQ/103001/2008, PTDC/DTP-FTO/ 0265/2012, and Pest-C/SAU/LA0001/2013-2014) and by the Norwegian Research Council, Project Number 190403. A.M.C., C.M.M, and A.L.C. are recipients of fellowships from the Portuguese Foundation for Science and Technology (SFRH/BD/63288/2009, SFRH/BD/79077/2011, and SFRH/BPD/46228/2008, respectively).por
dc.language.isoengpor
dc.publisherAmerican Chemical Societypor
dc.rightsopenAccesspor
dc.subjectsiRNA deliverypor
dc.subjectnonviral vectorpor
dc.subjectcancer gene therapypor
dc.subjectthermoresponsive copolymerpor
dc.subjectpolyplexpor
dc.subjectcloud pointpor
dc.titleApplication of Thermoresponsive PNIPAAM‑b‑PAMPTMA Diblock Copolymers in siRNA Deliverypor
dc.typearticlepor
degois.publication.firstPage819por
degois.publication.lastPage827por
degois.publication.issue3por
degois.publication.titleMolecular Pharmaceuticspor
dc.relation.publisherversionhttp://pubs.acs.org/doi/abs/10.1021/mp400510apor
dc.peerreviewedYespor
dc.identifier.doi10.1021/mp400510a-
degois.publication.volume11por
item.fulltextCom Texto completo-
item.grantfulltextopen-
item.languageiso639-1en-
crisitem.author.deptFaculty of Sciences and Technology-
crisitem.author.deptFaculty of Sciences and Technology-
crisitem.author.parentdeptUniversity of Coimbra-
crisitem.author.parentdeptUniversity of Coimbra-
crisitem.author.researchunitCQC - Coimbra Chemistry Centre-
crisitem.author.researchunitCNC - Center for Neuroscience and Cell Biology-
crisitem.author.researchunitCNC - Center for Neuroscience and Cell Biology-
crisitem.author.parentresearchunitFaculty of Sciences and Technology-
crisitem.author.orcid0000-0003-0551-7255-
crisitem.author.orcid0000-0003-1844-5027-
crisitem.author.orcid0000-0001-7095-5337-
Appears in Collections:FCTUC Ciências da Vida - Artigos em Revistas Internacionais
Files in This Item:
File Description SizeFormat
mp400510a.pdf2.77 MBAdobe PDFView/Open
Show simple item record

SCOPUSTM   
Citations

13
checked on Feb 18, 2020

WEB OF SCIENCETM
Citations 5

17
checked on Aug 2, 2022

Page view(s)

222
checked on Oct 4, 2022

Download(s)

287
checked on Oct 4, 2022

Google ScholarTM

Check

Altmetric

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.