Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralize
Autor(a) principal: | |
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Data de Publicação: | 2006 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da FURG (RI FURG) |
Texto Completo: | http://repositorio.furg.br/handle/1/3137 |
Resumo: | A plasmid expressing the â-galactosidase enzyme was used to transfect Vero cells in order to evaluate the efficiency of a liposome-mediated transfection by circular and linear DNA. The results obtained showed a low rate of transfection by linear DNA:liposome complexes. To explore whether the structure of the complexes was interfering with the transfection, atomic force microscopy (AFM) was used. It has confirmed the difference between the linear and circular condensates: whereas the circular DNA:liposome complexes presented compact spherical or cylindrical structures of about 100-800 nm, the linear DNA showed pearl necklace-like structures,with pearls varying from 250 to 400 nm. On the basis of the theory proposed by Kuhn et al. (1999), low concentrations of cationic amphihile were used to neutralize or reverse the DNA charge in order to improve the transfection efficiency of the linear DNA. Using this method,we were able to obtain the expression of the transgene without an associated toxicity observed with the linear DNA liposome delivery. |
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Groll, Andrea VonLevin, YanBarbosa, Marcia Cristina BernardesRavazzolo, Ana Paula2013-01-28T22:53:08Z2013-01-28T22:53:08Z2006GROLL, Andrea Von et al. Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralizer . Biotechnology v. 22, n.4, p. 1220-1224, 2006. Disponível em:<http://www.if.ufrgs.br/~levin/Pdfs.dir/biotech.pdf>. Acesso em:16 set. 2012.http://repositorio.furg.br/handle/1/3137A plasmid expressing the â-galactosidase enzyme was used to transfect Vero cells in order to evaluate the efficiency of a liposome-mediated transfection by circular and linear DNA. The results obtained showed a low rate of transfection by linear DNA:liposome complexes. To explore whether the structure of the complexes was interfering with the transfection, atomic force microscopy (AFM) was used. It has confirmed the difference between the linear and circular condensates: whereas the circular DNA:liposome complexes presented compact spherical or cylindrical structures of about 100-800 nm, the linear DNA showed pearl necklace-like structures,with pearls varying from 250 to 400 nm. On the basis of the theory proposed by Kuhn et al. (1999), low concentrations of cationic amphihile were used to neutralize or reverse the DNA charge in order to improve the transfection efficiency of the linear DNA. Using this method,we were able to obtain the expression of the transgene without an associated toxicity observed with the linear DNA liposome delivery.engLinear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralizeinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FURG (RI FURG)instname:Universidade Federal do Rio Grande (FURG)instacron:FURGORIGINALLinear DNA Low Efficiency Transfection by Liposome Can Be Improved by the Use of Cationic Lipid as Charge Neutralize.pdfLinear DNA Low Efficiency Transfection by Liposome Can Be Improved by the Use of Cationic Lipid as Charge Neutralize.pdfapplication/pdf371951https://repositorio.furg.br/bitstream/1/3137/1/Linear%20DNA%20Low%20Efficiency%20Transfection%20by%20Liposome%20Can%20Be%20Improved%20by%20the%20Use%20of%20Cationic%20Lipid%20as%20Charge%20Neutralize.pdf0a03e7add6e09a93bb9d608bdac0925aMD51open accessLICENSElicense.txtlicense.txttext/plain; charset=utf-81678https://repositorio.furg.br/bitstream/1/3137/2/license.txtd3be63d3b3eee02729709361dac69efeMD52open access1/31372013-01-28 20:53:08.659open accessoai:repositorio.furg.br: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ório InstitucionalPUBhttps://repositorio.furg.br/oai/request || http://200.19.254.174/oai/requestopendoar:2013-01-28T22:53:08Repositório Institucional da FURG (RI FURG) - Universidade Federal do Rio Grande (FURG)false |
dc.title.pt_BR.fl_str_mv |
Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralize |
title |
Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralize |
spellingShingle |
Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralize Groll, Andrea Von |
title_short |
Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralize |
title_full |
Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralize |
title_fullStr |
Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralize |
title_full_unstemmed |
Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralize |
title_sort |
Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralize |
author |
Groll, Andrea Von |
author_facet |
Groll, Andrea Von Levin, Yan Barbosa, Marcia Cristina Bernardes Ravazzolo, Ana Paula |
author_role |
author |
author2 |
Levin, Yan Barbosa, Marcia Cristina Bernardes Ravazzolo, Ana Paula |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Groll, Andrea Von Levin, Yan Barbosa, Marcia Cristina Bernardes Ravazzolo, Ana Paula |
description |
A plasmid expressing the â-galactosidase enzyme was used to transfect Vero cells in order to evaluate the efficiency of a liposome-mediated transfection by circular and linear DNA. The results obtained showed a low rate of transfection by linear DNA:liposome complexes. To explore whether the structure of the complexes was interfering with the transfection, atomic force microscopy (AFM) was used. It has confirmed the difference between the linear and circular condensates: whereas the circular DNA:liposome complexes presented compact spherical or cylindrical structures of about 100-800 nm, the linear DNA showed pearl necklace-like structures,with pearls varying from 250 to 400 nm. On the basis of the theory proposed by Kuhn et al. (1999), low concentrations of cationic amphihile were used to neutralize or reverse the DNA charge in order to improve the transfection efficiency of the linear DNA. Using this method,we were able to obtain the expression of the transgene without an associated toxicity observed with the linear DNA liposome delivery. |
publishDate |
2006 |
dc.date.issued.fl_str_mv |
2006 |
dc.date.accessioned.fl_str_mv |
2013-01-28T22:53:08Z |
dc.date.available.fl_str_mv |
2013-01-28T22:53:08Z |
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.citation.fl_str_mv |
GROLL, Andrea Von et al. Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralizer . Biotechnology v. 22, n.4, p. 1220-1224, 2006. Disponível em:<http://www.if.ufrgs.br/~levin/Pdfs.dir/biotech.pdf>. Acesso em:16 set. 2012. |
dc.identifier.uri.fl_str_mv |
http://repositorio.furg.br/handle/1/3137 |
identifier_str_mv |
GROLL, Andrea Von et al. Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralizer . Biotechnology v. 22, n.4, p. 1220-1224, 2006. Disponível em:<http://www.if.ufrgs.br/~levin/Pdfs.dir/biotech.pdf>. Acesso em:16 set. 2012. |
url |
http://repositorio.furg.br/handle/1/3137 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
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reponame:Repositório Institucional da FURG (RI FURG) instname:Universidade Federal do Rio Grande (FURG) instacron:FURG |
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Universidade Federal do Rio Grande (FURG) |
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FURG |
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FURG |
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Repositório Institucional da FURG (RI FURG) |
collection |
Repositório Institucional da FURG (RI FURG) |
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