Histidine Containing Peptides and Polypeptides as Nucleic Acid Vectors
Identifieur interne : 002224 ( Main/Exploration ); précédent : 002223; suivant : 002225Histidine Containing Peptides and Polypeptides as Nucleic Acid Vectors
Auteurs : Patrick Midoux [France] ; Eric Lecam [France] ; Dominique Coulaud [France] ; Etienne Delain [France] ; Chantal Pichon [France]Source :
- Somatic Cell and Molecular Genetics [ 0740-7750 ] ; 2002-11-01.
Abstract
Abstract: Nucleic acid transfer in mammalian cells is drastically improved with devices which increase their delivery in the cytosol upon endocytosis. In this chapter, we describe the effect on plasmid DNA (pDNA) and oligonucleotide (ODN) transfer, of an histidine-rich peptide (H5WYG), histidylated oligolysine (HoK), and histidylated polylysine (HpK) designed on the basis of the membrane destabilization capacity of poly-L-histidine at a pH close to that of the endosomes. We report that H5WYG, which permeabilizes the cell membrane at pH 6.4, favors the transfection mediated by lactosylated polylysine/pDNA complexes and, by lowering the pH of extracellular medium, allows the loading of the cytosol and the cell nucleus with ODN. We show that HoK forms small cationic spherical particles of 35 nm with ODN and HpK rod or toroid cationic particles of 100 nm with pDNA. PEGylation stabilizes these particles at physiological salt concentration. We also show that (i) HoK/ODN complexes yield a more than 20-fold increase of the biological activity of antisense ODN towards the inhibition of transient as well as constitutive gene expression and (ii) HpK/pDNA complexes yield a transfection efficiency of 3–4.5 order of magnitude higher than do polylysine/pDNA complexes. We also provide evidence that the effect of these polyhistidylated molecules is mediated by imidazole protonation in endosomes. Overall our data show that polyhistidylated molecules constitute interesting devices for an efficient cytosolic delivery of nucleic acids, and that ionic complexes between histidylated polylysine and a pDNA are attractive for developing a nonviral gene delivery system.
Url:
DOI: 10.1023/A:1022931923153
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type="city">Villejuif</settlement></placeName></affiliation></author><author><name sortKey="Coulaud, Dominique" sort="Coulaud, Dominique" uniqKey="Coulaud D" first="Dominique" last="Coulaud">Dominique Coulaud</name><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>Laboratoire de Microscopie Moléculaire et Cellulaire, UMR 8532 CNRS, Institut Gustave Roussy, rue Camille Desmoulins, F-94805, Villejuif cedex</wicri:regionArea><placeName><region type="region" nuts="2">Île-de-France</region><settlement type="city">Villejuif</settlement></placeName></affiliation></author><author><name sortKey="Delain, Etienne" sort="Delain, Etienne" uniqKey="Delain E" first="Etienne" last="Delain">Etienne Delain</name><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>Laboratoire de Microscopie Moléculaire et Cellulaire, UMR 8532 CNRS, Institut Gustave Roussy, rue Camille Desmoulins, F-94805, Villejuif 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Publishers</publisher><pubPlace>New York</pubPlace><date type="published" when="2002-11-01">2002-11-01</date><biblScope unit="volume">27</biblScope><biblScope unit="issue">1-6</biblScope><biblScope unit="page" from="27">27</biblScope><biblScope unit="page" to="47">47</biblScope></imprint><idno type="ISSN">0740-7750</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0740-7750</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Nucleic acid transfer in mammalian cells is drastically improved with devices which increase their delivery in the cytosol upon endocytosis. In this chapter, we describe the effect on plasmid DNA (pDNA) and oligonucleotide (ODN) transfer, of an histidine-rich peptide (H5WYG), histidylated oligolysine (HoK), and histidylated polylysine (HpK) designed on the basis of the membrane destabilization capacity of poly-L-histidine at a pH close to that of the endosomes. We report that H5WYG, which permeabilizes the cell membrane at pH 6.4, favors the transfection mediated by lactosylated polylysine/pDNA complexes and, by lowering the pH of extracellular medium, allows the loading of the cytosol and the cell nucleus with ODN. We show that HoK forms small cationic spherical particles of 35 nm with ODN and HpK rod or toroid cationic particles of 100 nm with pDNA. PEGylation stabilizes these particles at physiological salt concentration. We also show that (i) HoK/ODN complexes yield a more than 20-fold increase of the biological activity of antisense ODN towards the inhibition of transient as well as constitutive gene expression and (ii) HpK/pDNA complexes yield a transfection efficiency of 3–4.5 order of magnitude higher than do polylysine/pDNA complexes. We also provide evidence that the effect of these polyhistidylated molecules is mediated by imidazole protonation in endosomes. Overall our data show that polyhistidylated molecules constitute interesting devices for an efficient cytosolic delivery of nucleic acids, and that ionic complexes between histidylated polylysine and a pDNA are attractive for developing a nonviral gene delivery system.</div></front></TEI><affiliations><list><country><li>France</li></country><region><li>Centre-Val de Loire</li><li>Région Centre</li><li>Île-de-France</li></region><settlement><li>Orléans</li><li>Villejuif</li></settlement></list><tree><country name="France"><region name="Centre-Val de Loire"><name sortKey="Midoux, Patrick" sort="Midoux, Patrick" uniqKey="Midoux P" first="Patrick" last="Midoux">Patrick Midoux</name></region><name sortKey="Coulaud, Dominique" sort="Coulaud, Dominique" uniqKey="Coulaud D" first="Dominique" last="Coulaud">Dominique Coulaud</name><name sortKey="Delain, Etienne" sort="Delain, Etienne" uniqKey="Delain E" first="Etienne" last="Delain">Etienne Delain</name><name sortKey="Lecam, Eric" sort="Lecam, Eric" uniqKey="Lecam E" first="Eric" last="Lecam">Eric Lecam</name><name sortKey="Pichon, Chantal" sort="Pichon, Chantal" uniqKey="Pichon C" first="Chantal" last="Pichon">Chantal Pichon</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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