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DNA transfection and transfected cell viability using amphipathic asymmetric dendrimers

Identifieur interne : 002551 ( Main/Exploration ); précédent : 002550; suivant : 002552

DNA transfection and transfected cell viability using amphipathic asymmetric dendrimers

Auteurs : Dipen S. Shah [Royaume-Uni] ; T. Sakthivel [Royaume-Uni] ; Istvan Toth [Royaume-Uni] ; Alexander T. Florence [Royaume-Uni] ; Andy F. Wilderspin [Royaume-Uni]

Source :

International Journal of Pharmaceutics [ 0378-5173 ] ; 2000.

RBID : ISTEX:CD94E85ACB7135922B0A5CDE0F4AA6EF02D6BE35

English descriptors

Abstract

Abstract: Amphipathic asymmetric dendrimers have been investigated for use in delivery of genes into cells, with the objective of optimising transfection efficiency and maintaining cell viability. We have synthesised amphipathic asymmetric dendrimers by solid phase methods. The ability of two of these to transfect BHK cells in culture with β-galactosidase gene was determined by X-gal staining. Cell viability was measured by the MTT assay for BHK cells, and by spectroscopy for lysis of erythrocytes. Interactions between dendrimer and DNA were investigated by agarose gel electrophoresis. BHK cells were optimally transfected at 5:1 +/− charge ratio yielding 20% cells receiving at least one copy of the plasmid. Cell viability decreased when the dendrimer to DNA ratio exceeded 5:1. Raising the pH significantly affected the electrophoretic mobility of complexes of dendrimer and DNA. We conclude that amphipathic asymmetric dendrimers enable efficient plasmid DNA uptake into BHK cells. Cell viability is maintained at high concentrations of dendrimer when complexed with DNA at a 5:1 +/− charge ratio. Efficiency of transfection and cell viability suggest the system may be suitable for gene delivery in vivo.

Url:

https://api.istex.fr/ark:/67375/6H6-G5R0LGHP-C/fulltext.pdf

DOI: 10.1016/S0378-5173(00)00534-2

Affiliations:

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Le document en format XML

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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>DODAC, dioleyldimethylammonium chloride</term>
<term>DOGS, dioctadecylamidoglycylspermime</term>
<term>DOPE, dioleoyl phosphatidylethanolamine</term>
<term>DOSPA, 2,3-dioleyloxy-N-[2(Spermine-carboxamido)ethyl]-N,N-dimethyl-1-propanaminium trifluoracrtate</term>
<term>DOTMA, 2,3-diokeyloxypropyl-1-trimethyl ammonium chloride</term>
<term>Dendrimers</term>
<term>G9-EDA, 9th generation PAMAN with ethylenediamine core</term>
<term>Gene delivery</term>
<term>Gene expression</term>
<term>Non-viral vectors</term>
<term>PAMAM, polyamidoamine</term>
<term>Transfection</term>
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<keywords scheme="Teeft" xml:lang="en"><term>Agarose</term>
<term>Amphipathic</term>
<term>Aqueous solution</term>
<term>Brunswick square</term>
<term>Cationic</term>
<term>Cationic lipids</term>
<term>Cell viability</term>
<term>Charge ratio</term>
<term>Complex formation</term>
<term>Complex stability</term>
<term>Dendrimer</term>
<term>Dendrimer concentration</term>
<term>Dendrimers</term>
<term>Error bars show</term>
<term>Erythrocyte</term>
<term>Gene delivery</term>
<term>Gene expression</term>
<term>Gene transfer</term>
<term>Hydrophobic tail</term>
<term>International journal</term>
<term>Linear position</term>
<term>Lipid</term>
<term>Loading buffer</term>
<term>London wc1n</term>
<term>Mammalian cells</term>
<term>Minimal effect</term>
<term>Molar</term>
<term>Molar charge ratio</term>
<term>Molecular weight</term>
<term>Optimally transfected</term>
<term>Optimum charge ratio</term>
<term>Pamam</term>
<term>Pamam dendrimers</term>
<term>Percent survival</term>
<term>Percentage transfection</term>
<term>Pharmaceutics</term>
<term>Phosphate saline buffer</term>
<term>Plasmid</term>
<term>Psvbgal</term>
<term>Sakthivel</term>
<term>Shah</term>
<term>Stock solution</term>
<term>Supercoiled position</term>
<term>Terminal amines</term>
<term>Toth</term>
<term>Transfected</term>
<term>Transfection</term>
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<front><div type="abstract" xml:lang="en">Abstract: Amphipathic asymmetric dendrimers have been investigated for use in delivery of genes into cells, with the objective of optimising transfection efficiency and maintaining cell viability. We have synthesised amphipathic asymmetric dendrimers by solid phase methods. The ability of two of these to transfect BHK cells in culture with β-galactosidase gene was determined by X-gal staining. Cell viability was measured by the MTT assay for BHK cells, and by spectroscopy for lysis of erythrocytes. Interactions between dendrimer and DNA were investigated by agarose gel electrophoresis. BHK cells were optimally transfected at 5:1 +/− charge ratio yielding 20% cells receiving at least one copy of the plasmid. Cell viability decreased when the dendrimer to DNA ratio exceeded 5:1. Raising the pH significantly affected the electrophoretic mobility of complexes of dendrimer and DNA. We conclude that amphipathic asymmetric dendrimers enable efficient plasmid DNA uptake into BHK cells. Cell viability is maintained at high concentrations of dendrimer when complexed with DNA at a 5:1 +/− charge ratio. Efficiency of transfection and cell viability suggest the system may be suitable for gene delivery in vivo.</div>
</front>
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DNA transfection and transfected cell viability using amphipathic asymmetric dendrimers

DNA transfection and transfected cell viability using amphipathic asymmetric dendrimers

Source :

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri