Enhancement of nucleic acid delivery to hard-to-transfect human colorectal cancer cells by magnetofection at laminin coated substrates and promotion of the endosomal/lysosomal esca...

Autores
Cerda, Maria Belen; Batalla, Milena; Anton, Martina; Cafferata, Eduardo Gustavo Alfredo; Podhajcer, Osvaldo Luis; Plank, Christian; Mykhaylyk, Olga; Policastro, Lucia Laura
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Despite a great diversity of nanomaterials, such as cationic lipid, polymers or inorganic nanoparticles, has been developed in order to carry nucleic acids across plasma membrane, these methodologies have still insufficient efficacy in cells named hard-to-transfect cells, as the colorectal HT29 and Caco-2 cell lines. This paper describes the improvement of plasmid DNA (pDNA) and small interfering RNA (siRNA) transfer in these cells through the combination of magnetofection, a simplified extracellular matrix of laminin and endosomal/lysosomal escape promotion using the endosome-disruptive peptide INF-7. Magnetofection of pDNA complexes using selected vector formulations resulted in up to 2-fold enhancement in luciferase expression, as compared to lipofection. Further enhance in pDNA transfer in HT29 cells was obtained when magnetofection was applied on cells grown on laminin coated substrates, increasing 6-fold the luciferase expression compared to lipofection at uncoated substrates. This technique was also applied to siRNA delivery in cells expressing stably luciferase (Caco-2Luc and HT29Luc), selected magnetic vector formulations resulted in 61±5% and 50±5% of luciferase silencing in HT29Luc and Caco-2Luc, respectively. Further improvement in reporter gene silencing was obtained when the magnetic complexes were modified with INF-7, reaching more than 95% of luciferase silencing in Caco-2Luc cells, while pre-treatment of HT29Luc cells by chloroquine resulted in 80±4% of down regulation of luciferase expression. Thus, magnetofection applied on cells grown over laminin coated substrates and the optimization of endosomal escape of magnetic complexes would be a good alternative to enhance nucleic acid transfer in hard-to-transfect colorectal cancer cells.
Fil: Cerda, Maria Belen. Comision Nacional de Energia Atomica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Nanociencias y Nanotecnología; Argentina
Fil: Batalla, Milena. Comision Nacional de Energia Atomica; Argentina. Instituto de Nanociencias y Nanotecnología; Argentina
Fil: Anton, Martina. Technische Universität München; Alemania
Fil: Cafferata, Eduardo Gustavo Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Fundación Instituto Leloir; Argentina
Fil: Podhajcer, Osvaldo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Fundación Instituto Leloir; Argentina
Fil: Plank, Christian. Technische Universität München; Alemania
Fil: Mykhaylyk, Olga. Technische Universität München; Alemania
Fil: Policastro, Lucia Laura. Comision Nacional de Energia Atomica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Nanociencias y Nanotecnología; Argentina
Materia
Magnetic Nanoparticles
Magnetofection
Laminin
Inf-7
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/13821
Acceder
id CONICETDig_ad2cfc5e05f296ae708ddc18962d50de
oai_identifier_str oai:ri.conicet.gov.ar:11336/13821
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Enhancement of nucleic acid delivery to hard-to-transfect human colorectal cancer cells by magnetofection at laminin coated substrates and promotion of the endosomal/lysosomal escapeCerda, Maria BelenBatalla, MilenaAnton, MartinaCafferata, Eduardo Gustavo AlfredoPodhajcer, Osvaldo LuisPlank, ChristianMykhaylyk, OlgaPolicastro, Lucia LauraMagnetic NanoparticlesMagnetofectionLamininInf-7https://purl.org/becyt/ford/2.10https://purl.org/becyt/ford/2https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Despite a great diversity of nanomaterials, such as cationic lipid, polymers or inorganic nanoparticles, has been developed in order to carry nucleic acids across plasma membrane, these methodologies have still insufficient efficacy in cells named hard-to-transfect cells, as the colorectal HT29 and Caco-2 cell lines. This paper describes the improvement of plasmid DNA (pDNA) and small interfering RNA (siRNA) transfer in these cells through the combination of magnetofection, a simplified extracellular matrix of laminin and endosomal/lysosomal escape promotion using the endosome-disruptive peptide INF-7. Magnetofection of pDNA complexes using selected vector formulations resulted in up to 2-fold enhancement in luciferase expression, as compared to lipofection. Further enhance in pDNA transfer in HT29 cells was obtained when magnetofection was applied on cells grown on laminin coated substrates, increasing 6-fold the luciferase expression compared to lipofection at uncoated substrates. This technique was also applied to siRNA delivery in cells expressing stably luciferase (Caco-2Luc and HT29Luc), selected magnetic vector formulations resulted in 61±5% and 50±5% of luciferase silencing in HT29Luc and Caco-2Luc, respectively. Further improvement in reporter gene silencing was obtained when the magnetic complexes were modified with INF-7, reaching more than 95% of luciferase silencing in Caco-2Luc cells, while pre-treatment of HT29Luc cells by chloroquine resulted in 80±4% of down regulation of luciferase expression. Thus, magnetofection applied on cells grown over laminin coated substrates and the optimization of endosomal escape of magnetic complexes would be a good alternative to enhance nucleic acid transfer in hard-to-transfect colorectal cancer cells.Fil: Cerda, Maria Belen. Comision Nacional de Energia Atomica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Nanociencias y Nanotecnología; ArgentinaFil: Batalla, Milena. Comision Nacional de Energia Atomica; Argentina. Instituto de Nanociencias y Nanotecnología; ArgentinaFil: Anton, Martina. Technische Universität München; AlemaniaFil: Cafferata, Eduardo Gustavo Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Fundación Instituto Leloir; ArgentinaFil: Podhajcer, Osvaldo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Fundación Instituto Leloir; ArgentinaFil: Plank, Christian. Technische Universität München; AlemaniaFil: Mykhaylyk, Olga. Technische Universität München; AlemaniaFil: Policastro, Lucia Laura. Comision Nacional de Energia Atomica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Nanociencias y Nanotecnología; ArgentinaRoyal Society of Chemistry2015info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/13821Cerda, Maria Belen; Batalla, Milena; Anton, Martina; Cafferata, Eduardo Gustavo Alfredo; Podhajcer, Osvaldo Luis; et al.; Enhancement of nucleic acid delivery to hard-to-transfect human colorectal cancer cells by magnetofection at laminin coated substrates and promotion of the endosomal/lysosomal escape; Royal Society of Chemistry; RSC Advances; 72; 5; 2015; 58345-583542046-2069enginfo:eu-repo/semantics/altIdentifier/url/http://pubs.rsc.org/en/content/articlelanding/2015/ra/c5ra06562c#!divAbstractinfo:eu-repo/semantics/altIdentifier/url/http://dx.doi.org/10.1039/C5RA06562Cinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:21:12Zoai:ri.conicet.gov.ar:11336/13821instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:21:12.917CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Enhancement of nucleic acid delivery to hard-to-transfect human colorectal cancer cells by magnetofection at laminin coated substrates and promotion of the endosomal/lysosomal escape
title Enhancement of nucleic acid delivery to hard-to-transfect human colorectal cancer cells by magnetofection at laminin coated substrates and promotion of the endosomal/lysosomal escape
spellingShingle Enhancement of nucleic acid delivery to hard-to-transfect human colorectal cancer cells by magnetofection at laminin coated substrates and promotion of the endosomal/lysosomal escape
Cerda, Maria Belen
Magnetic Nanoparticles
Magnetofection
Laminin
Inf-7
title_short Enhancement of nucleic acid delivery to hard-to-transfect human colorectal cancer cells by magnetofection at laminin coated substrates and promotion of the endosomal/lysosomal escape
title_full Enhancement of nucleic acid delivery to hard-to-transfect human colorectal cancer cells by magnetofection at laminin coated substrates and promotion of the endosomal/lysosomal escape
title_fullStr Enhancement of nucleic acid delivery to hard-to-transfect human colorectal cancer cells by magnetofection at laminin coated substrates and promotion of the endosomal/lysosomal escape
title_full_unstemmed Enhancement of nucleic acid delivery to hard-to-transfect human colorectal cancer cells by magnetofection at laminin coated substrates and promotion of the endosomal/lysosomal escape
title_sort Enhancement of nucleic acid delivery to hard-to-transfect human colorectal cancer cells by magnetofection at laminin coated substrates and promotion of the endosomal/lysosomal escape
dc.creator.none.fl_str_mv Cerda, Maria Belen
Batalla, Milena
Anton, Martina
Cafferata, Eduardo Gustavo Alfredo
Podhajcer, Osvaldo Luis
Plank, Christian
Mykhaylyk, Olga
Policastro, Lucia Laura
author Cerda, Maria Belen
author_facet Cerda, Maria Belen
Batalla, Milena
Anton, Martina
Cafferata, Eduardo Gustavo Alfredo
Podhajcer, Osvaldo Luis
Plank, Christian
Mykhaylyk, Olga
Policastro, Lucia Laura
author_role author
author2 Batalla, Milena
Anton, Martina
Cafferata, Eduardo Gustavo Alfredo
Podhajcer, Osvaldo Luis
Plank, Christian
Mykhaylyk, Olga
Policastro, Lucia Laura
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Magnetic Nanoparticles
Magnetofection
Laminin
Inf-7
topic Magnetic Nanoparticles
Magnetofection
Laminin
Inf-7
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.10
https://purl.org/becyt/ford/2
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Despite a great diversity of nanomaterials, such as cationic lipid, polymers or inorganic nanoparticles, has been developed in order to carry nucleic acids across plasma membrane, these methodologies have still insufficient efficacy in cells named hard-to-transfect cells, as the colorectal HT29 and Caco-2 cell lines. This paper describes the improvement of plasmid DNA (pDNA) and small interfering RNA (siRNA) transfer in these cells through the combination of magnetofection, a simplified extracellular matrix of laminin and endosomal/lysosomal escape promotion using the endosome-disruptive peptide INF-7. Magnetofection of pDNA complexes using selected vector formulations resulted in up to 2-fold enhancement in luciferase expression, as compared to lipofection. Further enhance in pDNA transfer in HT29 cells was obtained when magnetofection was applied on cells grown on laminin coated substrates, increasing 6-fold the luciferase expression compared to lipofection at uncoated substrates. This technique was also applied to siRNA delivery in cells expressing stably luciferase (Caco-2Luc and HT29Luc), selected magnetic vector formulations resulted in 61±5% and 50±5% of luciferase silencing in HT29Luc and Caco-2Luc, respectively. Further improvement in reporter gene silencing was obtained when the magnetic complexes were modified with INF-7, reaching more than 95% of luciferase silencing in Caco-2Luc cells, while pre-treatment of HT29Luc cells by chloroquine resulted in 80±4% of down regulation of luciferase expression. Thus, magnetofection applied on cells grown over laminin coated substrates and the optimization of endosomal escape of magnetic complexes would be a good alternative to enhance nucleic acid transfer in hard-to-transfect colorectal cancer cells.
Fil: Cerda, Maria Belen. Comision Nacional de Energia Atomica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Nanociencias y Nanotecnología; Argentina
Fil: Batalla, Milena. Comision Nacional de Energia Atomica; Argentina. Instituto de Nanociencias y Nanotecnología; Argentina
Fil: Anton, Martina. Technische Universität München; Alemania
Fil: Cafferata, Eduardo Gustavo Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Fundación Instituto Leloir; Argentina
Fil: Podhajcer, Osvaldo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Fundación Instituto Leloir; Argentina
Fil: Plank, Christian. Technische Universität München; Alemania
Fil: Mykhaylyk, Olga. Technische Universität München; Alemania
Fil: Policastro, Lucia Laura. Comision Nacional de Energia Atomica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Nanociencias y Nanotecnología; Argentina
description Despite a great diversity of nanomaterials, such as cationic lipid, polymers or inorganic nanoparticles, has been developed in order to carry nucleic acids across plasma membrane, these methodologies have still insufficient efficacy in cells named hard-to-transfect cells, as the colorectal HT29 and Caco-2 cell lines. This paper describes the improvement of plasmid DNA (pDNA) and small interfering RNA (siRNA) transfer in these cells through the combination of magnetofection, a simplified extracellular matrix of laminin and endosomal/lysosomal escape promotion using the endosome-disruptive peptide INF-7. Magnetofection of pDNA complexes using selected vector formulations resulted in up to 2-fold enhancement in luciferase expression, as compared to lipofection. Further enhance in pDNA transfer in HT29 cells was obtained when magnetofection was applied on cells grown on laminin coated substrates, increasing 6-fold the luciferase expression compared to lipofection at uncoated substrates. This technique was also applied to siRNA delivery in cells expressing stably luciferase (Caco-2Luc and HT29Luc), selected magnetic vector formulations resulted in 61±5% and 50±5% of luciferase silencing in HT29Luc and Caco-2Luc, respectively. Further improvement in reporter gene silencing was obtained when the magnetic complexes were modified with INF-7, reaching more than 95% of luciferase silencing in Caco-2Luc cells, while pre-treatment of HT29Luc cells by chloroquine resulted in 80±4% of down regulation of luciferase expression. Thus, magnetofection applied on cells grown over laminin coated substrates and the optimization of endosomal escape of magnetic complexes would be a good alternative to enhance nucleic acid transfer in hard-to-transfect colorectal cancer cells.
publishDate 2015
dc.date.none.fl_str_mv 2015
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/13821
Cerda, Maria Belen; Batalla, Milena; Anton, Martina; Cafferata, Eduardo Gustavo Alfredo; Podhajcer, Osvaldo Luis; et al.; Enhancement of nucleic acid delivery to hard-to-transfect human colorectal cancer cells by magnetofection at laminin coated substrates and promotion of the endosomal/lysosomal escape; Royal Society of Chemistry; RSC Advances; 72; 5; 2015; 58345-58354
2046-2069
url http://hdl.handle.net/11336/13821
identifier_str_mv Cerda, Maria Belen; Batalla, Milena; Anton, Martina; Cafferata, Eduardo Gustavo Alfredo; Podhajcer, Osvaldo Luis; et al.; Enhancement of nucleic acid delivery to hard-to-transfect human colorectal cancer cells by magnetofection at laminin coated substrates and promotion of the endosomal/lysosomal escape; Royal Society of Chemistry; RSC Advances; 72; 5; 2015; 58345-58354
2046-2069
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://pubs.rsc.org/en/content/articlelanding/2015/ra/c5ra06562c#!divAbstract
info:eu-repo/semantics/altIdentifier/url/http://dx.doi.org/10.1039/C5RA06562C
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Royal Society of Chemistry
publisher.none.fl_str_mv Royal Society of Chemistry
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1844614199676764160
score 13.070432

Publicaciones similares