Histone H1-Mediated Transfection: Serum Inhibition Can Be Overcome by Ca2+ Ions
Identifieur interne : 002539 ( Main/Exploration ); précédent : 002538; suivant : 002540Histone H1-Mediated Transfection: Serum Inhibition Can Be Overcome by Ca2+ Ions
Auteurs : Annekathrin Haberland [Allemagne] ; Thomas Knaus [Allemagne] ; Sergei V. Zaitsev [Allemagne, Russie] ; Bernd Buchberger [Allemagne] ; Andreas Lun [Allemagne] ; Hermann Haller [Allemagne] ; Michael Böttger [Allemagne]Source :
- Pharmaceutical Research [ 0724-8741 ] ; 2000-02-01.
English descriptors
- KwdEn :
Abstract
Abstract: Purpose. One of the drawbacks of polycationic and cationic liposomalgene transfer is its sensitivity to serum. Gene therapy requires thetransfectant-DNA complex to be resistant to serum as well as blood.Since Ca2+ has proved to be an efficient cofactor of polycationic genetransfer, we decided to investigate its effects on transfection in thepresence of serum. Methods. We studied transgene expression of luciferase gene (pCMVLuc) on ECV 304 human endothelial cells using H1 histone andDOSPER as transfectants in the presence of 0-100% fetal calf serum. Results. H1-and DOSPER-mediated transfection was found to beinhibited by serum above the concentration of 10%. If 2 mM Ca2+ or2 mM Ca2+/0.1 mM chloroquine was included in the culture mediumwhich replace the transfection mixture and was left on the cells for24 hours postincubation, the inhibiting effect of even 100% serumwas overcome. Conclusions. A high serum level does not interfere with binding anduptake of H1- and DOSPER-DNA complexes, but inhibits subsequentsteps such as endosomal escape. Ca2+ in the form of nascent calciumphosphate microprecipitates and other lysosomolytical agents facilitateendosomal/lysosomal release by their fusigenic and membranolyticactivity.
Url:
DOI: 10.1023/A:1007581700996
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 000515
- to stream Istex, to step Curation: 000515
- to stream Istex, to step Checkpoint: 001357
- to stream Main, to step Merge: 002567
- to stream Main, to step Curation: 002539
Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Histone H1-Mediated Transfection: Serum Inhibition Can Be Overcome by Ca2+ Ions</title><author><name sortKey="Haberland, Annekathrin" sort="Haberland, Annekathrin" uniqKey="Haberland A" first="Annekathrin" last="Haberland">Annekathrin Haberland</name></author><author><name sortKey="Knaus, Thomas" sort="Knaus, Thomas" uniqKey="Knaus T" first="Thomas" last="Knaus">Thomas Knaus</name></author><author><name sortKey="Zaitsev, Sergei V" sort="Zaitsev, Sergei V" uniqKey="Zaitsev S" first="Sergei V." last="Zaitsev">Sergei V. Zaitsev</name></author><author><name sortKey="Buchberger, Bernd" sort="Buchberger, Bernd" uniqKey="Buchberger B" first="Bernd" last="Buchberger">Bernd Buchberger</name></author><author><name sortKey="Lun, Andreas" sort="Lun, Andreas" uniqKey="Lun A" first="Andreas" last="Lun">Andreas Lun</name></author><author><name sortKey="Haller, Hermann" sort="Haller, Hermann" uniqKey="Haller H" first="Hermann" last="Haller">Hermann Haller</name></author><author><name sortKey="Bottger, Michael" sort="Bottger, Michael" uniqKey="Bottger M" first="Michael" last="Böttger">Michael Böttger</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:5E39256AF6D067F01D2FA923B6BCC3574FA0A9AD</idno><date when="2000" year="2000">2000</date><idno type="doi">10.1023/A:1007581700996</idno><idno type="url">https://api.istex.fr/ark:/67375/VQC-Z8N2MRJ5-4/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">000515</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000515</idno><idno type="wicri:Area/Istex/Curation">000515</idno><idno type="wicri:Area/Istex/Checkpoint">001357</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">001357</idno><idno type="wicri:doubleKey">0724-8741:2000:Haberland A:histone:h:mediated</idno><idno type="wicri:Area/Main/Merge">002567</idno><idno type="wicri:Area/Main/Curation">002539</idno><idno type="wicri:Area/Main/Exploration">002539</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Histone H1-Mediated Transfection: Serum Inhibition Can Be Overcome by Ca2+ Ions</title><author><name sortKey="Haberland, Annekathrin" sort="Haberland, Annekathrin" uniqKey="Haberland A" first="Annekathrin" last="Haberland">Annekathrin Haberland</name><affiliation wicri:level="1"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Max Delbrück Center for Molecular Medicine, Franz Volhard Clinic, Berlin-Buch</wicri:regionArea><wicri:noRegion>Berlin-Buch</wicri:noRegion><wicri:noRegion>Berlin-Buch</wicri:noRegion></affiliation></author><author><name sortKey="Knaus, Thomas" sort="Knaus, Thomas" uniqKey="Knaus T" first="Thomas" last="Knaus">Thomas Knaus</name><affiliation wicri:level="1"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Max Delbrück Center for Molecular Medicine, Franz Volhard Clinic, Berlin-Buch</wicri:regionArea><wicri:noRegion>Berlin-Buch</wicri:noRegion><wicri:noRegion>Berlin-Buch</wicri:noRegion></affiliation></author><author><name sortKey="Zaitsev, Sergei V" sort="Zaitsev, Sergei V" uniqKey="Zaitsev S" first="Sergei V." last="Zaitsev">Sergei V. Zaitsev</name><affiliation wicri:level="1"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Max Delbrück Center for Molecular Medicine, Franz Volhard Clinic, Berlin-Buch</wicri:regionArea><wicri:noRegion>Berlin-Buch</wicri:noRegion><wicri:noRegion>Berlin-Buch</wicri:noRegion></affiliation><affiliation wicri:level="1"><country xml:lang="fr">Russie</country><wicri:regionArea>Institute of Cytology of the Russian Academy of Sciences, St. Petersburg</wicri:regionArea><wicri:noRegion>St. Petersburg</wicri:noRegion></affiliation></author><author><name sortKey="Buchberger, Bernd" sort="Buchberger, Bernd" uniqKey="Buchberger B" first="Bernd" last="Buchberger">Bernd Buchberger</name><affiliation wicri:level="1"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Roche Diagnostics GmbH, Penzberg</wicri:regionArea><wicri:noRegion>Penzberg</wicri:noRegion><wicri:noRegion>Penzberg</wicri:noRegion></affiliation></author><author><name sortKey="Lun, Andreas" sort="Lun, Andreas" uniqKey="Lun A" first="Andreas" last="Lun">Andreas Lun</name><affiliation wicri:level="3"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Institute of Laboratory Medicine and Pathobiochemistry, Charité, Humboldt University, Berlin</wicri:regionArea><placeName><region type="land" nuts="3">Berlin</region><settlement type="city">Berlin</settlement></placeName></affiliation></author><author><name sortKey="Haller, Hermann" sort="Haller, Hermann" uniqKey="Haller H" first="Hermann" last="Haller">Hermann Haller</name><affiliation wicri:level="1"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Max Delbrück Center for Molecular Medicine, Franz Volhard Clinic, Berlin-Buch</wicri:regionArea><wicri:noRegion>Berlin-Buch</wicri:noRegion><wicri:noRegion>Berlin-Buch</wicri:noRegion></affiliation></author><author><name sortKey="Bottger, Michael" sort="Bottger, Michael" uniqKey="Bottger M" first="Michael" last="Böttger">Michael Böttger</name><affiliation wicri:level="1"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Max Delbrück Center for Molecular Medicine, Franz Volhard Clinic, Berlin-Buch</wicri:regionArea><wicri:noRegion>Berlin-Buch</wicri:noRegion><wicri:noRegion>Berlin-Buch</wicri:noRegion></affiliation><affiliation wicri:level="3"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Institute of Laboratory Medicine and Pathobiochemistry, Charité, Humboldt University, Berlin</wicri:regionArea><placeName><region type="land" nuts="3">Berlin</region><settlement type="city">Berlin</settlement></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j">Pharmaceutical Research</title><title level="j" type="sub">An Official Journal of the American Association of Pharmaceutical Scientists</title><title level="j" type="abbrev">Pharm Res</title><idno type="ISSN">0724-8741</idno><idno type="eISSN">1573-904X</idno><imprint><publisher>Kluwer Academic Publishers-Plenum Publishers</publisher><pubPlace>New York</pubPlace><date type="published" when="2000-02-01">2000-02-01</date><biblScope unit="volume">17</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="229">229</biblScope><biblScope unit="page" to="235">235</biblScope></imprint><idno type="ISSN">0724-8741</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0724-8741</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>DOSPER</term><term>H1 histone</term><term>calcium</term><term>serum inhibition</term><term>transfection</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Purpose. One of the drawbacks of polycationic and cationic liposomalgene transfer is its sensitivity to serum. Gene therapy requires thetransfectant-DNA complex to be resistant to serum as well as blood.Since Ca2+ has proved to be an efficient cofactor of polycationic genetransfer, we decided to investigate its effects on transfection in thepresence of serum. Methods. We studied transgene expression of luciferase gene (pCMVLuc) on ECV 304 human endothelial cells using H1 histone andDOSPER as transfectants in the presence of 0-100% fetal calf serum. Results. H1-and DOSPER-mediated transfection was found to beinhibited by serum above the concentration of 10%. If 2 mM Ca2+ or2 mM Ca2+/0.1 mM chloroquine was included in the culture mediumwhich replace the transfection mixture and was left on the cells for24 hours postincubation, the inhibiting effect of even 100% serumwas overcome. Conclusions. A high serum level does not interfere with binding anduptake of H1- and DOSPER-DNA complexes, but inhibits subsequentsteps such as endosomal escape. Ca2+ in the form of nascent calciumphosphate microprecipitates and other lysosomolytical agents facilitateendosomal/lysosomal release by their fusigenic and membranolyticactivity.</div></front></TEI><affiliations><list><country><li>Allemagne</li><li>Russie</li></country><region><li>Berlin</li></region><settlement><li>Berlin</li></settlement></list><tree><country name="Allemagne"><noRegion><name sortKey="Haberland, Annekathrin" sort="Haberland, Annekathrin" uniqKey="Haberland A" first="Annekathrin" last="Haberland">Annekathrin Haberland</name></noRegion><name sortKey="Bottger, Michael" sort="Bottger, Michael" uniqKey="Bottger M" first="Michael" last="Böttger">Michael Böttger</name><name sortKey="Bottger, Michael" sort="Bottger, Michael" uniqKey="Bottger M" first="Michael" last="Böttger">Michael Böttger</name><name sortKey="Buchberger, Bernd" sort="Buchberger, Bernd" uniqKey="Buchberger B" first="Bernd" last="Buchberger">Bernd Buchberger</name><name sortKey="Haller, Hermann" sort="Haller, Hermann" uniqKey="Haller H" first="Hermann" last="Haller">Hermann Haller</name><name sortKey="Knaus, Thomas" sort="Knaus, Thomas" uniqKey="Knaus T" first="Thomas" last="Knaus">Thomas Knaus</name><name sortKey="Lun, Andreas" sort="Lun, Andreas" uniqKey="Lun A" first="Andreas" last="Lun">Andreas Lun</name><name sortKey="Zaitsev, Sergei V" sort="Zaitsev, Sergei V" uniqKey="Zaitsev S" first="Sergei V." last="Zaitsev">Sergei V. Zaitsev</name></country><country name="Russie"><noRegion><name sortKey="Zaitsev, Sergei V" sort="Zaitsev, Sergei V" uniqKey="Zaitsev S" first="Sergei V." last="Zaitsev">Sergei V. Zaitsev</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/ChloroquineV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 002539 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 002539 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= ChloroquineV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= ISTEX:5E39256AF6D067F01D2FA923B6BCC3574FA0A9AD
|texte= Histone H1-Mediated Transfection: Serum Inhibition Can Be Overcome by Ca2+ Ions
}}
| This area was generated with Dilib version V0.6.33. |  |