Structural dynamics and physicochemical properties of pDNA/DODAB:MO lipoplexes : effect of pH and anionic lipids in inverted non-lamellar phases versus lamellar phases
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/1822/29738 |
Resumo: | Dioctadecyldimethylammoniumbromide (DODAB):Monoolein (MO) lipoplexes have mainly been studied within the range of high molar ratios of DODAB, with noticeable transfection efficiencies in the Human Embryonic Kidney (HEK, a.k.a. 293T) cell line. In thiswork,we intend to study the effect of highMOcontent on the structure and physicochemical properties of pDNA/DODAB:MOlipoplexes to achieve some correlationwith their transfection efficiency. Static/Dynamic Light Scattering and Cryo-TEM imaging were used to characterize the size/ morphology of DNA/DODAB:MO lipoplexes at different DODAB:MO contents (2:1, 1:1, 1:2) and charge ratios (CRs) (+/−). Nile Red fluorescence emission was performed to detect changes in microviscosity, hydration and polarity of DNA/DODAB:MO systems. Lipoplexes stability at physiological pH values and in the presence of anionic lipids was evaluated by Förster Resonance Energy Transfer (FRET). Physicochemical/structural data were complemented with transfection studies in HEK cells using the β-galactosidase reporter gene activity assay. This work reports the coexistence of multilamellar and non-lamellar inverted phases in MO-richer lipoplexes (DODAB:MO 1:2 and 1:4), leading to transfection efficiencies comparable to those of multilamellar (DODAB-richer) lipoplexes, but at higher charge ratios [CR (+/−) = 6.0] and without dose-effect response. These results may be related to the structural changes of lipoplexes promoted by high MO content. |
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Structural dynamics and physicochemical properties of pDNA/DODAB:MO lipoplexes : effect of pH and anionic lipids in inverted non-lamellar phases versus lamellar phasesLipoplexesTransfectionMonooleinCryo-TEMFRETLight scatteringNile RedScience & TechnologyDioctadecyldimethylammoniumbromide (DODAB):Monoolein (MO) lipoplexes have mainly been studied within the range of high molar ratios of DODAB, with noticeable transfection efficiencies in the Human Embryonic Kidney (HEK, a.k.a. 293T) cell line. In thiswork,we intend to study the effect of highMOcontent on the structure and physicochemical properties of pDNA/DODAB:MOlipoplexes to achieve some correlationwith their transfection efficiency. Static/Dynamic Light Scattering and Cryo-TEM imaging were used to characterize the size/ morphology of DNA/DODAB:MO lipoplexes at different DODAB:MO contents (2:1, 1:1, 1:2) and charge ratios (CRs) (+/−). Nile Red fluorescence emission was performed to detect changes in microviscosity, hydration and polarity of DNA/DODAB:MO systems. Lipoplexes stability at physiological pH values and in the presence of anionic lipids was evaluated by Förster Resonance Energy Transfer (FRET). Physicochemical/structural data were complemented with transfection studies in HEK cells using the β-galactosidase reporter gene activity assay. This work reports the coexistence of multilamellar and non-lamellar inverted phases in MO-richer lipoplexes (DODAB:MO 1:2 and 1:4), leading to transfection efficiencies comparable to those of multilamellar (DODAB-richer) lipoplexes, but at higher charge ratios [CR (+/−) = 6.0] and without dose-effect response. These results may be related to the structural changes of lipoplexes promoted by high MO content.FEDER (037291) through POFC — COMPETEFundação para a Ciência e a Tecnologia (FCT) - SFRH/BD/46968/2009 (PhD grant), PEst-C/BIA/UI4050/2011(CBMA), PEst-C/FIS/UI0607/2011 (CFUM).ElsevierUniversidade do MinhoSilva, João P. N.Oliveira, I. M. S. C.Oliveira, Ana Cristina Norberto GonçalvesLúcio, M.Gomes, A. C.Coutinho, Paulo J. G.Oliveira, M. Elisabete2014-06-272014-06-27T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/29738eng0005-273610.1016/j.bbamem.2014.06.01424976292www.elsevier.com/locate/bbameminfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T12:53:50Zoai:repositorium.sdum.uminho.pt:1822/29738Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:53:19.666186Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Structural dynamics and physicochemical properties of pDNA/DODAB:MO lipoplexes : effect of pH and anionic lipids in inverted non-lamellar phases versus lamellar phases |
title |
Structural dynamics and physicochemical properties of pDNA/DODAB:MO lipoplexes : effect of pH and anionic lipids in inverted non-lamellar phases versus lamellar phases |
spellingShingle |
Structural dynamics and physicochemical properties of pDNA/DODAB:MO lipoplexes : effect of pH and anionic lipids in inverted non-lamellar phases versus lamellar phases Silva, João P. N. Lipoplexes Transfection Monoolein Cryo-TEM FRET Light scattering Nile Red Science & Technology |
title_short |
Structural dynamics and physicochemical properties of pDNA/DODAB:MO lipoplexes : effect of pH and anionic lipids in inverted non-lamellar phases versus lamellar phases |
title_full |
Structural dynamics and physicochemical properties of pDNA/DODAB:MO lipoplexes : effect of pH and anionic lipids in inverted non-lamellar phases versus lamellar phases |
title_fullStr |
Structural dynamics and physicochemical properties of pDNA/DODAB:MO lipoplexes : effect of pH and anionic lipids in inverted non-lamellar phases versus lamellar phases |
title_full_unstemmed |
Structural dynamics and physicochemical properties of pDNA/DODAB:MO lipoplexes : effect of pH and anionic lipids in inverted non-lamellar phases versus lamellar phases |
title_sort |
Structural dynamics and physicochemical properties of pDNA/DODAB:MO lipoplexes : effect of pH and anionic lipids in inverted non-lamellar phases versus lamellar phases |
author |
Silva, João P. N. |
author_facet |
Silva, João P. N. Oliveira, I. M. S. C. Oliveira, Ana Cristina Norberto Gonçalves Lúcio, M. Gomes, A. C. Coutinho, Paulo J. G. Oliveira, M. Elisabete |
author_role |
author |
author2 |
Oliveira, I. M. S. C. Oliveira, Ana Cristina Norberto Gonçalves Lúcio, M. Gomes, A. C. Coutinho, Paulo J. G. Oliveira, M. Elisabete |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade do Minho |
dc.contributor.author.fl_str_mv |
Silva, João P. N. Oliveira, I. M. S. C. Oliveira, Ana Cristina Norberto Gonçalves Lúcio, M. Gomes, A. C. Coutinho, Paulo J. G. Oliveira, M. Elisabete |
dc.subject.por.fl_str_mv |
Lipoplexes Transfection Monoolein Cryo-TEM FRET Light scattering Nile Red Science & Technology |
topic |
Lipoplexes Transfection Monoolein Cryo-TEM FRET Light scattering Nile Red Science & Technology |
description |
Dioctadecyldimethylammoniumbromide (DODAB):Monoolein (MO) lipoplexes have mainly been studied within the range of high molar ratios of DODAB, with noticeable transfection efficiencies in the Human Embryonic Kidney (HEK, a.k.a. 293T) cell line. In thiswork,we intend to study the effect of highMOcontent on the structure and physicochemical properties of pDNA/DODAB:MOlipoplexes to achieve some correlationwith their transfection efficiency. Static/Dynamic Light Scattering and Cryo-TEM imaging were used to characterize the size/ morphology of DNA/DODAB:MO lipoplexes at different DODAB:MO contents (2:1, 1:1, 1:2) and charge ratios (CRs) (+/−). Nile Red fluorescence emission was performed to detect changes in microviscosity, hydration and polarity of DNA/DODAB:MO systems. Lipoplexes stability at physiological pH values and in the presence of anionic lipids was evaluated by Förster Resonance Energy Transfer (FRET). Physicochemical/structural data were complemented with transfection studies in HEK cells using the β-galactosidase reporter gene activity assay. This work reports the coexistence of multilamellar and non-lamellar inverted phases in MO-richer lipoplexes (DODAB:MO 1:2 and 1:4), leading to transfection efficiencies comparable to those of multilamellar (DODAB-richer) lipoplexes, but at higher charge ratios [CR (+/−) = 6.0] and without dose-effect response. These results may be related to the structural changes of lipoplexes promoted by high MO content. |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014-06-27 2014-06-27T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/1822/29738 |
url |
http://hdl.handle.net/1822/29738 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
0005-2736 10.1016/j.bbamem.2014.06.014 24976292 www.elsevier.com/locate/bbamem |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Elsevier |
publisher.none.fl_str_mv |
Elsevier |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799133129304178688 |