Protein expression knockdown in cancer cells induced by a gemini cationic lipid nanovector with histidine-based polar heads

A histidine-based gemini cationic lipid, which had already demonstrated its efficiency as a plasmid DNA (pDNA) nanocarrier, has been used in this work to transfect a small interfering RNA (siRNA) into cancer cells. In combination with the helper lipid monoolein glycerol (MOG), the cationic lipid was...

ver descrição completa

Detalhes bibliográficos
Autores: Sánchez Arribas, Natalia, Martínez Negro, María, Villar; Eva M., Pérez, Lourdes, Osío Barcina, José De Jesús, Aicart Sospedra, Emilio, Taboada, Pablo, Guerrero Martínez, Andrés, Junquera González, María Elena
Formato: artículo
Fecha de publicación:2020
País:España
Recursos:Universidad Complutense de Madrid (UCM)
Repositorio:Docta Complutense
Idioma:inglés
OAI Identifier:oai:docta.ucm.es:20.500.14352/108924
Acesso em linha:https://hdl.handle.net/20.500.14352/108924
Access Level:acceso abierto
Palavra-chave:544
577.1
Non-viral gene delivery
Gene knockdown efficiency
Small interfering RNA
Amino acid-based gemini cationic lipids
Protein expression
Protein corona
Química física (Química)
Bioquímica (Química)
2307 Química Física
2302.26 Bioquímica Física
id ES_38db079e93a7a9541fbd3618c06e297c
oai_identifier_str oai:docta.ucm.es:20.500.14352/108924
network_acronym_str ES
network_name_str España
repository_id_str
spelling Protein expression knockdown in cancer cells induced by a gemini cationic lipid nanovector with histidine-based polar headsSánchez Arribas, NataliaMartínez Negro, MaríaVillar; Eva M.Pérez, LourdesOsío Barcina, José De JesúsAicart Sospedra, EmilioTaboada, PabloGuerrero Martínez, AndrésJunquera González, María Elena544577.1Non-viral gene deliveryGene knockdown efficiencySmall interfering RNAAmino acid-based gemini cationic lipidsProtein expressionProtein coronaQuímica física (Química)Bioquímica (Química)2307 Química Física2302.26 Bioquímica FísicaA histidine-based gemini cationic lipid, which had already demonstrated its efficiency as a plasmid DNA (pDNA) nanocarrier, has been used in this work to transfect a small interfering RNA (siRNA) into cancer cells. In combination with the helper lipid monoolein glycerol (MOG), the cationic lipid was used as an antiGFP-siRNA nanovector in a multidisciplinary study. Initially, a biophysical characterization by zeta potential and agarose gel electrophoresis experiments was performed to determine the lipid effective charge and confirm siRNA compaction. The lipoplexes formed were arranged in Lalfa lamellar lyotropic liquid crystal phases with a cluster-type morphology, as cryo-transmission electron microscopy (cryo-TEM) and small-angle X-ray scattering (SAXS) studies revealed. Additionally, in vitro experiments confirmed the high gene knockdown efficiency of the lipid-based nanovehicle as detected by flow cytometry (FC) and epifluorescence microscopy, even better than that of Lipofectamine2000*, the transfecting reagent commonly used as a positive control. Cytotoxicity assays indicated that the nanovector is non-toxic to cells. Finally, using nano-liquid chromatography tandem mass spectrometry (nanoLC-MS/MS), apolipoprotein A-I and A-II followed by serum albumin were identified as the proteins with higher affinity for the surface of the lipoplexes. This fact could be beyond the remarkable silencing activity of the histidine-based lipid nanocarrier herein presented.MDPIUniversidad Complutense de Madrid20202020-01-0120202020-01-01journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/20.500.14352/108924reponame:Docta Complutenseinstname:Universidad Complutense de Madrid (UCM)InglésengMinisterio de Ciencia e Innovación http://dx.doi.org/10.13039/501100004837 Not available RTI2018-095844-B-I00Ministerio de Educación y Ciencia Not available CTQ2017-88948-PMinisterio de Educación y Ciencia Not available UCMA05-33-010Ministerio de Educación y Ciencia Not available MAT2016-80266-Ropen accesshttp://purl.org/coar/access_right/c_abf2Attribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessoai:docta.ucm.es:20.500.14352/1089242026-06-02T12:44:21Z
dc.title.none.fl_str_mv Protein expression knockdown in cancer cells induced by a gemini cationic lipid nanovector with histidine-based polar heads
title Protein expression knockdown in cancer cells induced by a gemini cationic lipid nanovector with histidine-based polar heads
spellingShingle Protein expression knockdown in cancer cells induced by a gemini cationic lipid nanovector with histidine-based polar heads
Sánchez Arribas, Natalia
544
577.1
Non-viral gene delivery
Gene knockdown efficiency
Small interfering RNA
Amino acid-based gemini cationic lipids
Protein expression
Protein corona
Química física (Química)
Bioquímica (Química)
2307 Química Física
2302.26 Bioquímica Física
title_short Protein expression knockdown in cancer cells induced by a gemini cationic lipid nanovector with histidine-based polar heads
title_full Protein expression knockdown in cancer cells induced by a gemini cationic lipid nanovector with histidine-based polar heads
title_fullStr Protein expression knockdown in cancer cells induced by a gemini cationic lipid nanovector with histidine-based polar heads
title_full_unstemmed Protein expression knockdown in cancer cells induced by a gemini cationic lipid nanovector with histidine-based polar heads
title_sort Protein expression knockdown in cancer cells induced by a gemini cationic lipid nanovector with histidine-based polar heads
dc.creator.none.fl_str_mv Sánchez Arribas, Natalia
Martínez Negro, María
Villar; Eva M.
Pérez, Lourdes
Osío Barcina, José De Jesús
Aicart Sospedra, Emilio
Taboada, Pablo
Guerrero Martínez, Andrés
Junquera González, María Elena
author Sánchez Arribas, Natalia
author_facet Sánchez Arribas, Natalia
Martínez Negro, María
Villar; Eva M.
Pérez, Lourdes
Osío Barcina, José De Jesús
Aicart Sospedra, Emilio
Taboada, Pablo
Guerrero Martínez, Andrés
Junquera González, María Elena
author_role author
author2 Martínez Negro, María
Villar; Eva M.
Pérez, Lourdes
Osío Barcina, José De Jesús
Aicart Sospedra, Emilio
Taboada, Pablo
Guerrero Martínez, Andrés
Junquera González, María Elena
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidad Complutense de Madrid
dc.subject.none.fl_str_mv 544
577.1
Non-viral gene delivery
Gene knockdown efficiency
Small interfering RNA
Amino acid-based gemini cationic lipids
Protein expression
Protein corona
Química física (Química)
Bioquímica (Química)
2307 Química Física
2302.26 Bioquímica Física
topic 544
577.1
Non-viral gene delivery
Gene knockdown efficiency
Small interfering RNA
Amino acid-based gemini cationic lipids
Protein expression
Protein corona
Química física (Química)
Bioquímica (Química)
2307 Química Física
2302.26 Bioquímica Física
description A histidine-based gemini cationic lipid, which had already demonstrated its efficiency as a plasmid DNA (pDNA) nanocarrier, has been used in this work to transfect a small interfering RNA (siRNA) into cancer cells. In combination with the helper lipid monoolein glycerol (MOG), the cationic lipid was used as an antiGFP-siRNA nanovector in a multidisciplinary study. Initially, a biophysical characterization by zeta potential and agarose gel electrophoresis experiments was performed to determine the lipid effective charge and confirm siRNA compaction. The lipoplexes formed were arranged in Lalfa lamellar lyotropic liquid crystal phases with a cluster-type morphology, as cryo-transmission electron microscopy (cryo-TEM) and small-angle X-ray scattering (SAXS) studies revealed. Additionally, in vitro experiments confirmed the high gene knockdown efficiency of the lipid-based nanovehicle as detected by flow cytometry (FC) and epifluorescence microscopy, even better than that of Lipofectamine2000*, the transfecting reagent commonly used as a positive control. Cytotoxicity assays indicated that the nanovector is non-toxic to cells. Finally, using nano-liquid chromatography tandem mass spectrometry (nanoLC-MS/MS), apolipoprotein A-I and A-II followed by serum albumin were identified as the proteins with higher affinity for the surface of the lipoplexes. This fact could be beyond the remarkable silencing activity of the histidine-based lipid nanocarrier herein presented.
publishDate 2020
dc.date.none.fl_str_mv 2020
2020-01-01
2020
2020-01-01
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/20.500.14352/108924
url https://hdl.handle.net/20.500.14352/108924
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv Ministerio de Ciencia e Innovación http://dx.doi.org/10.13039/501100004837 Not available RTI2018-095844-B-I00
Ministerio de Educación y Ciencia Not available CTQ2017-88948-P
Ministerio de Educación y Ciencia Not available UCMA05-33-010
Ministerio de Educación y Ciencia Not available MAT2016-80266-R
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:Docta Complutense
instname:Universidad Complutense de Madrid (UCM)
instname_str Universidad Complutense de Madrid (UCM)
reponame_str Docta Complutense
collection Docta Complutense
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869406128291446784
score 15.198674

Registos relacionados