Efficient Gene Transfer by Histidylated Polylysine/pDNA Complexes
Identifieur interne : 000719 ( Istex/Curation ); précédent : 000718; suivant : 000720Efficient Gene Transfer by Histidylated Polylysine/pDNA Complexes
Auteurs : Patrick Midoux [France] ; Michel Monsigny [France]Source :
- Bioconjugate Chemistry [ 1043-1802 ] ; 1999.
Abstract
Plasmid/polylysine complexes, which are used to transfect mammalian cells, increase the uptake of DNA, but plasmid molecules are sequestered into vesicles where they cannot escape to reach the nuclear machinery. However, the transfection efficiency increases when membrane-disrupting reagents such as chloroquine or fusogenic peptides, are used to disrupt endosomal membranes and to favor the delivery of plasmid into the cytosol. We designed a cationic polymer that forms complexes with a plasmid DNA (pDNA) and mediates the transfection of various cell lines in the absence of chloroquine or fusogenic peptides. This polymer is a polylysine (average degree of polymerization of 190) partially substituted with histidyl residues which become cationic upon protonation of the imidazole groups at pH below 6.0. The transfection efficiency was optimal with a polylysine having 38 ± 5% of the ε-amino groups substituted with histidyl residues; it was not significantly impaired in the presence of serum in the culture medium. The transfection was drastically inhibited in the presence of bafilomycin A1, indicating that the protonation of the imidazole groups in the endosome lumen might favor the delivery of pDNA into the cytosol.
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DOI: 10.1021/bc9801070
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<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title>Efficient Gene Transfer by Histidylated Polylysine/pDNA Complexes</title><author><name sortKey="Midoux, Patrick" sort="Midoux, Patrick" uniqKey="Midoux P" first="Patrick" last="Midoux">Patrick Midoux</name><affiliation wicri:level="1"><mods:affiliation>Centre de Biophysique Moléculaire, Glycobiologie CNRS UPR4301 and University of Orléans,rue Charles-Sadron, F-45071 Orléans Cedex 02, France</mods:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Centre de Biophysique Moléculaire, Glycobiologie CNRS UPR4301 and University of Orléans,rue Charles-Sadron, F-45071 Orléans Cedex 02</wicri:regionArea></affiliation><affiliation wicri:level="1"><mods:affiliation> To whom correspondence should be addressed. Fax: 33 (0)238 69 00 94. Phone: 38 (0)2 38 25 55 95. E-mail: midoux@cnrs-orleans.fr.</mods:affiliation><country wicri:rule="url">France</country></affiliation></author><author><name sortKey="Monsigny, Michel" sort="Monsigny, Michel" uniqKey="Monsigny M" first="Michel" last="Monsigny">Michel Monsigny</name><affiliation wicri:level="1"><mods:affiliation>Centre de Biophysique Moléculaire, Glycobiologie CNRS UPR4301 and University of Orléans,rue Charles-Sadron, F-45071 Orléans Cedex 02, France</mods:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Centre de Biophysique Moléculaire, Glycobiologie CNRS UPR4301 and University of Orléans,rue Charles-Sadron, F-45071 Orléans Cedex 02</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:78F597C5553A58D20F360CE80E382DEB873465D7</idno><date when="1999" year="1999">1999</date><idno type="doi">10.1021/bc9801070</idno><idno type="url">https://api.istex.fr/ark:/67375/TPS-N3J84NW4-S/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">000719</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000719</idno><idno type="wicri:Area/Istex/Curation">000719</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Efficient Gene Transfer by Histidylated Polylysine/pDNA Complexes</title><author><name sortKey="Midoux, Patrick" sort="Midoux, Patrick" uniqKey="Midoux P" first="Patrick" last="Midoux">Patrick Midoux</name><affiliation wicri:level="1"><mods:affiliation>Centre de Biophysique Moléculaire, Glycobiologie CNRS UPR4301 and University of Orléans,rue Charles-Sadron, F-45071 Orléans Cedex 02, France</mods:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Centre de Biophysique Moléculaire, Glycobiologie CNRS UPR4301 and University of Orléans,rue Charles-Sadron, F-45071 Orléans Cedex 02</wicri:regionArea></affiliation><affiliation wicri:level="1"><mods:affiliation> To whom correspondence should be addressed. Fax: 33 (0)238 69 00 94. Phone: 38 (0)2 38 25 55 95. E-mail: midoux@cnrs-orleans.fr.</mods:affiliation><country wicri:rule="url">France</country></affiliation></author><author><name sortKey="Monsigny, Michel" sort="Monsigny, Michel" uniqKey="Monsigny M" first="Michel" last="Monsigny">Michel Monsigny</name><affiliation wicri:level="1"><mods:affiliation>Centre de Biophysique Moléculaire, Glycobiologie CNRS UPR4301 and University of Orléans,rue Charles-Sadron, F-45071 Orléans Cedex 02, France</mods:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Centre de Biophysique Moléculaire, Glycobiologie CNRS UPR4301 and University of Orléans,rue Charles-Sadron, F-45071 Orléans Cedex 02</wicri:regionArea></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">Bioconjugate Chemistry</title><title level="j" type="abbrev">Bioconjugate Chem.</title><idno type="ISSN">1043-1802</idno><idno type="eISSN">1520-4812</idno><imprint><publisher>American Chemical Society</publisher><date type="e-published" when="1999-03-11">1999</date><date when="1999-05-17">1999</date><biblScope unit="vol">10</biblScope><biblScope unit="issue">3</biblScope><biblScope unit="page" from="406">406</biblScope><biblScope unit="page" to="411">411</biblScope></imprint><idno type="ISSN">1043-1802</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">1043-1802</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract">Plasmid/polylysine complexes, which are used to transfect mammalian cells, increase the uptake of DNA, but plasmid molecules are sequestered into vesicles where they cannot escape to reach the nuclear machinery. However, the transfection efficiency increases when membrane-disrupting reagents such as chloroquine or fusogenic peptides, are used to disrupt endosomal membranes and to favor the delivery of plasmid into the cytosol. We designed a cationic polymer that forms complexes with a plasmid DNA (pDNA) and mediates the transfection of various cell lines in the absence of chloroquine or fusogenic peptides. This polymer is a polylysine (average degree of polymerization of 190) partially substituted with histidyl residues which become cationic upon protonation of the imidazole groups at pH below 6.0. The transfection efficiency was optimal with a polylysine having 38 ± 5% of the ε-amino groups substituted with histidyl residues; it was not significantly impaired in the presence of serum in the culture medium. The transfection was drastically inhibited in the presence of bafilomycin A1, indicating that the protonation of the imidazole groups in the endosome lumen might favor the delivery of pDNA into the cytosol.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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