In Vitro Transfection Mediated by Dendrigraft Poly(L‐lysines): The Effect of Structure and Molecule Size
Identifieur interne : 002136 ( Main/Exploration ); précédent : 002135; suivant : 002137In Vitro Transfection Mediated by Dendrigraft Poly(L‐lysines): The Effect of Structure and Molecule Size
Auteurs : Jakub Hofman [République tchèque] ; Martin Buncek [République tchèque] ; Radovan Haluza [République tchèque] ; Ludvik Streinz [République tchèque] ; Miroslav Ledvina [République tchèque] ; Petr Cigler [République tchèque]Source :
- Macromolecular Bioscience [ 1616-5187 ] ; 2013-02.
English descriptors
- Teeft :
- Amine, Assay, Automatic shaker, Better comparison, Biosci, Buncek, Cell debris, Cell lysate, Cell viability, Cellular experiments, Chem, Cigler, Commercial transfection reagent, Control transfection, Cytotoxicity, Czech, Czech republic, Dendrigraft, Dendrimers, Dendritic, Dendritic polymers, Dnase, Dropwise addition, Drug delivery, Egfp, Eppendorf tubes, Gmbh, Growth medium, Haluza, Hep2, Hep2 cells, Hep2_egfp cells, High cytotoxicity, Highest concentration, Hofman, Hradec kralove, Incubation period, Independent experiments, Internalization, Jetpei, Kgaa, Ledvina, Lipofectamine, Lowest value, Luciferase, Lysate, Macromol, Maximal, Maximal effect, Molecular weight, Montpellier cedex, Nuclease, Oligonucleotide, Other experiments, Other hand, Pci_luc, Pharm, Physiological saline, Plasmid, Plasmid pci_luc, Plasmid transfection, Polymer, Polyplexes, Protein assay, Protein content, Proton sponge mechanism, Pure lysate, Reagent, Room temperature, Same amount, Same time, Several optimalizations, Sirna, Standard conditions, Streinz, Thermal cycler, Transfection, Transfection abilities, Transfection experiments, Transfection procedure, Verlag, Verlag gmbh, Weinheim.
Abstract
Dendritic poly(L‐lysines) (DGL) constitute promising nanomaterials applicable as a nonviral gene‐delivery vector. In this study, we evaluate the transfection abilities of four DGL generations with special emphasis on the systematic description of the relationship of how generation (i.e., molecule size) affects the transfection efficacy. Using Hep2 cells, we demonstrated that the capability of unmodified DGL to deliver plasmid is of a magnitude lower than that of jetPEI. On the other hand, employing the Hep2 cell line stably transduced with eGFP, we observed that DGL G5 delivers the siRNA oligonucleotide with the same efficiency as Lipofectamine 2000. In further experiments, it was shown that DGL affords excellent ability to bind DNA, protect it against DNase I attack, and internalize it into cells.
The relationship between the molecule size of dendrigraft poly(L‐lysines) (DGL) and their transfection abilities is described systematically. The DGL transfection capability for oligonucleotide is much higher than for plasmid, while generations G3, G4, and G5 are more effective transfection mediators in comparison with G2. A possible structure‐based explanation for the transfection behavior of DGL is given.
Url:
DOI: 10.1002/mabi.201200303
Affiliations:
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Czech Republic, Flemingovo namesti 2, 166 10 Prague 6</wicri:regionArea><wicri:noRegion>166 10 Prague 6</wicri:noRegion></affiliation></author><author><name sortKey="Ledvina, Miroslav" sort="Ledvina, Miroslav" uniqKey="Ledvina M" first="Miroslav" last="Ledvina">Miroslav Ledvina</name><affiliation wicri:level="1"><country xml:lang="fr">République tchèque</country><wicri:regionArea>Institute of Organic Chemistry and Biochemistry, v.v.i., Academy of Sciences of the Czech Republic, Flemingovo namesti 2, 166 10 Prague 6</wicri:regionArea><wicri:noRegion>166 10 Prague 6</wicri:noRegion></affiliation></author><author><name sortKey="Cigler, Petr" sort="Cigler, Petr" uniqKey="Cigler P" first="Petr" last="Cigler">Petr Cigler</name><affiliation wicri:level="1"><country xml:lang="fr">République tchèque</country><wicri:regionArea>Institute of Organic Chemistry and Biochemistry, v.v.i., Academy of Sciences of the Czech Republic, Flemingovo namesti 2, 166 10 Prague 6</wicri:regionArea><wicri:noRegion>166 10 Prague 6</wicri:noRegion></affiliation><affiliation wicri:level="1"><country wicri:rule="url">République tchèque</country></affiliation><affiliation wicri:level="1"><country xml:lang="fr">République tchèque</country><wicri:regionArea>Correspondence address: Institute of Organic Chemistry and Biochemistry, v.v.i., Academy of Sciences of the Czech Republic, Flemingovo namesti 2, 166 10 Prague 6</wicri:regionArea><wicri:noRegion>166 10 Prague 6</wicri:noRegion></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">Macromolecular Bioscience</title><title level="j" type="alt">MACROMOLECULAR BIOSCIENCE</title><idno type="ISSN">1616-5187</idno><idno type="eISSN">1616-5195</idno><imprint><biblScope unit="vol">13</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="167">167</biblScope><biblScope unit="page" to="176">176</biblScope><biblScope unit="page-count">10</biblScope><publisher>WILEY‐VCH Verlag</publisher><pubPlace>Weinheim</pubPlace><date type="published" when="2013-02">2013-02</date></imprint><idno type="ISSN">1616-5187</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">1616-5187</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Amine</term><term>Assay</term><term>Automatic shaker</term><term>Better comparison</term><term>Biosci</term><term>Buncek</term><term>Cell debris</term><term>Cell lysate</term><term>Cell viability</term><term>Cellular experiments</term><term>Chem</term><term>Cigler</term><term>Commercial transfection reagent</term><term>Control transfection</term><term>Cytotoxicity</term><term>Czech</term><term>Czech republic</term><term>Dendrigraft</term><term>Dendrimers</term><term>Dendritic</term><term>Dendritic polymers</term><term>Dnase</term><term>Dropwise addition</term><term>Drug delivery</term><term>Egfp</term><term>Eppendorf tubes</term><term>Gmbh</term><term>Growth medium</term><term>Haluza</term><term>Hep2</term><term>Hep2 cells</term><term>Hep2_egfp cells</term><term>High cytotoxicity</term><term>Highest concentration</term><term>Hofman</term><term>Hradec kralove</term><term>Incubation period</term><term>Independent experiments</term><term>Internalization</term><term>Jetpei</term><term>Kgaa</term><term>Ledvina</term><term>Lipofectamine</term><term>Lowest value</term><term>Luciferase</term><term>Lysate</term><term>Macromol</term><term>Maximal</term><term>Maximal effect</term><term>Molecular weight</term><term>Montpellier cedex</term><term>Nuclease</term><term>Oligonucleotide</term><term>Other experiments</term><term>Other hand</term><term>Pci_luc</term><term>Pharm</term><term>Physiological saline</term><term>Plasmid</term><term>Plasmid pci_luc</term><term>Plasmid transfection</term><term>Polymer</term><term>Polyplexes</term><term>Protein assay</term><term>Protein content</term><term>Proton sponge mechanism</term><term>Pure lysate</term><term>Reagent</term><term>Room temperature</term><term>Same amount</term><term>Same time</term><term>Several optimalizations</term><term>Sirna</term><term>Standard conditions</term><term>Streinz</term><term>Thermal cycler</term><term>Transfection</term><term>Transfection abilities</term><term>Transfection experiments</term><term>Transfection procedure</term><term>Verlag</term><term>Verlag gmbh</term><term>Weinheim</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Dendritic poly(L‐lysines) (DGL) constitute promising nanomaterials applicable as a nonviral gene‐delivery vector. In this study, we evaluate the transfection abilities of four DGL generations with special emphasis on the systematic description of the relationship of how generation (i.e., molecule size) affects the transfection efficacy. Using Hep2 cells, we demonstrated that the capability of unmodified DGL to deliver plasmid is of a magnitude lower than that of jetPEI. On the other hand, employing the Hep2 cell line stably transduced with eGFP, we observed that DGL G5 delivers the siRNA oligonucleotide with the same efficiency as Lipofectamine 2000. In further experiments, it was shown that DGL affords excellent ability to bind DNA, protect it against DNase I attack, and internalize it into cells.</div><div type="abstract" xml:lang="en">The relationship between the molecule size of dendrigraft poly(L‐lysines) (DGL) and their transfection abilities is described systematically. The DGL transfection capability for oligonucleotide is much higher than for plasmid, while generations G3, G4, and G5 are more effective transfection mediators in comparison with G2. A possible structure‐based explanation for the transfection behavior of DGL is given.</div></front></TEI><affiliations><list><country><li>République tchèque</li></country></list><tree><country name="République tchèque"><noRegion><name sortKey="Hofman, Jakub" sort="Hofman, Jakub" uniqKey="Hofman J" first="Jakub" last="Hofman">Jakub Hofman</name></noRegion><name sortKey="Buncek, Martin" sort="Buncek, Martin" uniqKey="Buncek M" first="Martin" last="Buncek">Martin Buncek</name><name sortKey="Cigler, Petr" sort="Cigler, Petr" uniqKey="Cigler P" first="Petr" last="Cigler">Petr Cigler</name><name sortKey="Cigler, Petr" sort="Cigler, Petr" uniqKey="Cigler P" first="Petr" last="Cigler">Petr Cigler</name><name sortKey="Cigler, Petr" sort="Cigler, Petr" uniqKey="Cigler P" first="Petr" last="Cigler">Petr Cigler</name><name sortKey="Haluza, Radovan" sort="Haluza, Radovan" uniqKey="Haluza R" first="Radovan" last="Haluza">Radovan Haluza</name><name sortKey="Hofman, Jakub" sort="Hofman, Jakub" uniqKey="Hofman J" first="Jakub" last="Hofman">Jakub Hofman</name><name sortKey="Ledvina, Miroslav" sort="Ledvina, Miroslav" uniqKey="Ledvina M" first="Miroslav" last="Ledvina">Miroslav Ledvina</name><name sortKey="Streinz, Ludvik" sort="Streinz, Ludvik" uniqKey="Streinz L" first="Ludvik" last="Streinz">Ludvik Streinz</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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