Vectors based on reducible polycations facilitate intracellular release of nucleic acids
Identifieur interne : 000C04 ( Istex/Curation ); précédent : 000C03; suivant : 000C05Vectors based on reducible polycations facilitate intracellular release of nucleic acids
Auteurs : Martin L. Read [Royaume-Uni] ; K. Helen Bremner [Royaume-Uni] ; David Oupick [Royaume-Uni] ; Nicola K. Green [Royaume-Uni] ; Peter F. Searle [Royaume-Uni] ; Leonard W. Seymour [Royaume-Uni]Source :
- The Journal of Gene Medicine [ 1099-498X ] ; 2003-03.
English descriptors
- Teeft :
- Assay, Bioconjug chem, Cancer cells, Cancer gene, Cancer gene therapy, Cationic, Cationic lipid, Cationic lipids, Cell lines, Cells transfected, Cellular uptake, Chem, Chloroquine, Copyright, Cytometry analysis, Cytosolic nucleases, Dotap, Endosomal, Ethidium, Ethidium bromide, Expression plasmid, Fresh media, Gene, Gene delivery, Gene expression, Gene therapy, Gene transfer, Grey bars, Hela, Hela cells, Hepes buffer, High levels, Higher level, Imagequanttm software, Initial experiments, Intracellular, Intracellular activation, Intracellular delivery, Intracellular release, John wiley sons, Lipid, Lipoplexes, Lncap, Lncap cells, Luciferase, Luciferase activity, Molecular weight, Mrna, Nacl, Nitroreductase, Nuclease, Nuclease degradation, Nucleic, Nucleic acids, Oxidative polycondensation, Pegfpn1, Peptide, Photon correlation spectroscopy, Physiological conditions, Plasmid, Polycation, Polycations, Polyplexes, Prodrug, Reducible, Reducible polycations, Reporter gene expression, Similar levels, Standard deviation, Surface charge, Synthetic vectors, Toxicity, Transfected, Transfected cells, Transfection, Transfection activity, Uorescence, White bars.
Abstract
Background: Inefficient intracellular delivery of nucleic acids limits the therapeutic usefulness of synthetic vectors such as poly(L‐lysine) (PLL)/DNA polyplexes. This article reports on the characterisation of a new type of synthetic vector based on a linear reducible polycation (RPC) that can be cleaved by the intracellular environment to facilitate release of nucleic acids. Methods: RPCs of molecular weight (mwt) 45 and 187 kDa were prepared by oxidative polycondensation of the peptide Cys‐Lys10‐Cys and used to condense nucleic acids. The stability of RPC‐based polyplexes to reduction was determined using electrophoresis, dynamic light scattering and fluorescence techniques. Transfection activity was studied in several cancer cell lines (HeLa, LNCaP, PC‐3 and B16‐F10) using luciferase and green fluorescent protein (GFP) genes as reporter genes in the presence of chloroquine or the cationic lipid (N‐(1‐(2,3‐dioleoyloxy)propyl)‐N, N, N‐trimethylammonium chloride) (DOTAP). A CMV‐driven plasmid expressing the nitroreductase (ntr) gene was used to evaluate the therapeutic efficacy of RPC‐based delivery vectors. Results: A 187‐fold higher level of gene expression indicated that intracellular delivery of DNA was more efficient using RPC/DOTAP compared with vectors based on non‐reducible PLL. Analysis by flow cytometry also showed enhanced delivery of the GFP gene by RPC/DOTAP in HeLa (51.5 ± 7.9%), LNCaP (55.2 ± 6.7%) and PC‐3 (66.1 ± 3.7%) cells. Transfection with the ntr gene and treatment with the prodrug CB1954 resulted in significant cell killing, achieving IC50 values similar to those previously attained with adenoviral vectors. Delivery of mRNA (20–75% of cells) was also more efficient using RPC/DOTAP than PLL/DOTAP (<5% of cells). Conclusions: These results demonstrate that lipid‐mediated activation of RPC‐based polyplexes is a useful strategy to enhance intracellular delivery of nucleic acids and potentiate therapeutic activity. Copyright © 2002 John Wiley & Sons, Ltd.
Url:
DOI: 10.1002/jgm.331
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: Pour aller vers cette notice dans l'étape Curation :000C04
Links to Exploration step
ISTEX:5B64D673FF4FBAD3BE0BB46200FF07964DD46B8CLe document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Vectors based on reducible polycations facilitate intracellular release of nucleic acids</title><author><name sortKey="Read, Martin L" sort="Read, Martin L" uniqKey="Read M" first="Martin L." last="Read">Martin L. Read</name><affiliation wicri:level="1"><mods:affiliation>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT, UK</mods:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT</wicri:regionArea></affiliation><affiliation wicri:level="1"><mods:affiliation>E-mail: mread@globalnet.co.uk</mods:affiliation><country wicri:rule="url">Royaume-Uni</country></affiliation><affiliation wicri:level="1"><mods:affiliation>Correspondence address: Department of Clinical Pharmacology, University of Oxford, Oxford OX2 6HE, UK.</mods:affiliation><country xml:lang="fr" wicri:curation="lc">Royaume-Uni</country><wicri:regionArea>Correspondence address: Department of Clinical Pharmacology, University of Oxford, Oxford OX2 6HE</wicri:regionArea></affiliation></author><author><name sortKey="Bremner, K Helen" sort="Bremner, K Helen" uniqKey="Bremner K" first="K. Helen" last="Bremner">K. Helen Bremner</name><affiliation wicri:level="1"><mods:affiliation>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT, UK</mods:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT</wicri:regionArea></affiliation></author><author><name sortKey="Oupick, David" sort="Oupick, David" uniqKey="Oupick D" first="David" last="Oupick">David Oupick</name><affiliation wicri:level="1"><mods:affiliation>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT, UK</mods:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT</wicri:regionArea></affiliation></author><author><name sortKey="Green, Nicola K" sort="Green, Nicola K" uniqKey="Green N" first="Nicola K." last="Green">Nicola K. Green</name><affiliation wicri:level="1"><mods:affiliation>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT, UK</mods:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT</wicri:regionArea></affiliation></author><author><name sortKey="Searle, Peter F" sort="Searle, Peter F" uniqKey="Searle P" first="Peter F." last="Searle">Peter F. Searle</name><affiliation wicri:level="1"><mods:affiliation>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT, UK</mods:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT</wicri:regionArea></affiliation></author><author><name sortKey="Seymour, Leonard W" sort="Seymour, Leonard W" uniqKey="Seymour L" first="Leonard W." last="Seymour">Leonard W. Seymour</name><affiliation wicri:level="1"><mods:affiliation>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT, UK</mods:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:5B64D673FF4FBAD3BE0BB46200FF07964DD46B8C</idno><date when="2003" year="2003">2003</date><idno type="doi">10.1002/jgm.331</idno><idno type="url">https://api.istex.fr/ark:/67375/WNG-PHJ5C7S9-J/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">000C04</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000C04</idno><idno type="wicri:Area/Istex/Curation">000C04</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main">Vectors based on reducible polycations facilitate intracellular release of nucleic acids</title><author><name sortKey="Read, Martin L" sort="Read, Martin L" uniqKey="Read M" first="Martin L." last="Read">Martin L. Read</name><affiliation wicri:level="1"><mods:affiliation>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT, UK</mods:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT</wicri:regionArea></affiliation><affiliation wicri:level="1"><mods:affiliation>E-mail: mread@globalnet.co.uk</mods:affiliation><country wicri:rule="url">Royaume-Uni</country></affiliation><affiliation wicri:level="1"><mods:affiliation>Correspondence address: Department of Clinical Pharmacology, University of Oxford, Oxford OX2 6HE, UK.</mods:affiliation><country xml:lang="fr" wicri:curation="lc">Royaume-Uni</country><wicri:regionArea>Correspondence address: Department of Clinical Pharmacology, University of Oxford, Oxford OX2 6HE</wicri:regionArea></affiliation></author><author><name sortKey="Bremner, K Helen" sort="Bremner, K Helen" uniqKey="Bremner K" first="K. Helen" last="Bremner">K. Helen Bremner</name><affiliation wicri:level="1"><mods:affiliation>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT, UK</mods:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT</wicri:regionArea></affiliation></author><author><name sortKey="Oupick, David" sort="Oupick, David" uniqKey="Oupick D" first="David" last="Oupick">David Oupick</name><affiliation wicri:level="1"><mods:affiliation>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT, UK</mods:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT</wicri:regionArea></affiliation></author><author><name sortKey="Green, Nicola K" sort="Green, Nicola K" uniqKey="Green N" first="Nicola K." last="Green">Nicola K. Green</name><affiliation wicri:level="1"><mods:affiliation>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT, UK</mods:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT</wicri:regionArea></affiliation></author><author><name sortKey="Searle, Peter F" sort="Searle, Peter F" uniqKey="Searle P" first="Peter F." last="Searle">Peter F. Searle</name><affiliation wicri:level="1"><mods:affiliation>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT, UK</mods:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT</wicri:regionArea></affiliation></author><author><name sortKey="Seymour, Leonard W" sort="Seymour, Leonard W" uniqKey="Seymour L" first="Leonard W." last="Seymour">Leonard W. Seymour</name><affiliation wicri:level="1"><mods:affiliation>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT, UK</mods:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT</wicri:regionArea></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">The Journal of Gene Medicine</title><title level="j" type="alt">JOURNAL OF GENE MEDICINE, THE</title><idno type="ISSN">1099-498X</idno><idno type="eISSN">1521-2254</idno><imprint><biblScope unit="vol">5</biblScope><biblScope unit="issue">3</biblScope><biblScope unit="page" from="232">232</biblScope><biblScope unit="page" to="245">245</biblScope><biblScope unit="page-count">14</biblScope><publisher>John Wiley & Sons, Ltd.</publisher><pubPlace>Chichester, UK</pubPlace><date type="published" when="2003-03">2003-03</date></imprint><idno type="ISSN">1099-498X</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">1099-498X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Assay</term><term>Bioconjug chem</term><term>Cancer cells</term><term>Cancer gene</term><term>Cancer gene therapy</term><term>Cationic</term><term>Cationic lipid</term><term>Cationic lipids</term><term>Cell lines</term><term>Cells transfected</term><term>Cellular uptake</term><term>Chem</term><term>Chloroquine</term><term>Copyright</term><term>Cytometry analysis</term><term>Cytosolic nucleases</term><term>Dotap</term><term>Endosomal</term><term>Ethidium</term><term>Ethidium bromide</term><term>Expression plasmid</term><term>Fresh media</term><term>Gene</term><term>Gene delivery</term><term>Gene expression</term><term>Gene therapy</term><term>Gene transfer</term><term>Grey bars</term><term>Hela</term><term>Hela cells</term><term>Hepes buffer</term><term>High levels</term><term>Higher level</term><term>Imagequanttm software</term><term>Initial experiments</term><term>Intracellular</term><term>Intracellular activation</term><term>Intracellular delivery</term><term>Intracellular release</term><term>John wiley sons</term><term>Lipid</term><term>Lipoplexes</term><term>Lncap</term><term>Lncap cells</term><term>Luciferase</term><term>Luciferase activity</term><term>Molecular weight</term><term>Mrna</term><term>Nacl</term><term>Nitroreductase</term><term>Nuclease</term><term>Nuclease degradation</term><term>Nucleic</term><term>Nucleic acids</term><term>Oxidative polycondensation</term><term>Pegfpn1</term><term>Peptide</term><term>Photon correlation spectroscopy</term><term>Physiological conditions</term><term>Plasmid</term><term>Polycation</term><term>Polycations</term><term>Polyplexes</term><term>Prodrug</term><term>Reducible</term><term>Reducible polycations</term><term>Reporter gene expression</term><term>Similar levels</term><term>Standard deviation</term><term>Surface charge</term><term>Synthetic vectors</term><term>Toxicity</term><term>Transfected</term><term>Transfected cells</term><term>Transfection</term><term>Transfection activity</term><term>Uorescence</term><term>White bars</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Background: Inefficient intracellular delivery of nucleic acids limits the therapeutic usefulness of synthetic vectors such as poly(L‐lysine) (PLL)/DNA polyplexes. This article reports on the characterisation of a new type of synthetic vector based on a linear reducible polycation (RPC) that can be cleaved by the intracellular environment to facilitate release of nucleic acids. Methods: RPCs of molecular weight (mwt) 45 and 187 kDa were prepared by oxidative polycondensation of the peptide Cys‐Lys10‐Cys and used to condense nucleic acids. The stability of RPC‐based polyplexes to reduction was determined using electrophoresis, dynamic light scattering and fluorescence techniques. Transfection activity was studied in several cancer cell lines (HeLa, LNCaP, PC‐3 and B16‐F10) using luciferase and green fluorescent protein (GFP) genes as reporter genes in the presence of chloroquine or the cationic lipid (N‐(1‐(2,3‐dioleoyloxy)propyl)‐N, N, N‐trimethylammonium chloride) (DOTAP). A CMV‐driven plasmid expressing the nitroreductase (ntr) gene was used to evaluate the therapeutic efficacy of RPC‐based delivery vectors. Results: A 187‐fold higher level of gene expression indicated that intracellular delivery of DNA was more efficient using RPC/DOTAP compared with vectors based on non‐reducible PLL. Analysis by flow cytometry also showed enhanced delivery of the GFP gene by RPC/DOTAP in HeLa (51.5 ± 7.9%), LNCaP (55.2 ± 6.7%) and PC‐3 (66.1 ± 3.7%) cells. Transfection with the ntr gene and treatment with the prodrug CB1954 resulted in significant cell killing, achieving IC50 values similar to those previously attained with adenoviral vectors. Delivery of mRNA (20–75% of cells) was also more efficient using RPC/DOTAP than PLL/DOTAP (<5% of cells). Conclusions: These results demonstrate that lipid‐mediated activation of RPC‐based polyplexes is a useful strategy to enhance intracellular delivery of nucleic acids and potentiate therapeutic activity. Copyright © 2002 John Wiley & Sons, Ltd.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/ChloroquineV1/Data/Istex/Curation
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000C04 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Istex/Curation/biblio.hfd -nk 000C04 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= ChloroquineV1
|flux= Istex
|étape= Curation
|type= RBID
|clé= ISTEX:5B64D673FF4FBAD3BE0BB46200FF07964DD46B8C
|texte= Vectors based on reducible polycations facilitate intracellular release of nucleic acids
}}
| This area was generated with Dilib version V0.6.33. |  |