Polypropylenimine generation four: a suitable vector for targeted gene alteration in vitro

Journal of Drug Targeting
Juergen Klingler, Dieter Kaufmann

Abstract

Polypropylenimine dendrimers have been shown to be alternative vectors for DNA delivery. Up to now, polypropylenimine dendrimers have not been investigated for the transport of specific single stranded modified oligonucleotides for targeted gene alteration. We investigated generation four polypropylenimine dendrimers to target a 45 base (phosphorothioate modified) DNA oligonucleotide in the cell nuclei of cultured cells in order to correct a specific point mutation in hprt. Transfection resulted in a high cellular uptake of the FITC-labeled oligonucleotides in V79 hamster fibroblasts and HuH-7 human liver cell lines, at low toxicity levels. Using transmission electron microscopy and immunolabeling, oligonucleotides were found 24 h after transfection predominantly in the nuclear compartment. The oligonucleotides showed the desired biological activity, the correction of the hprt point mutation in V79-400 cells. For the transfection of modified oligonucleotides in targeted gene alteration, generation four polypropylenimine dendrimer is a suitable vector.

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