title: Polypropylenimine generation four: a suitable vector for targeted gene alteration in vitro.
abstract: BACKGROUND
  Polypropylenimine dendrimers have been shown to be alternative vectors for DNA delivery.
  
  
  OBJECTIVE
  Up to now, polypropylenimine dendrimers have not been investigated for the transport of specific single stranded modified oligonucleotides for targeted gene alteration.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  For the transfection of modified oligonucleotides in targeted gene alteration, generation four polypropylenimine dendrimer is a suitable vector.
authors.affiliation: Institute of Human Genetics, University of Ulm, Albert-Einstein-Allee, Ulm, Germany.
authors.forename: Juergen
authors.initials: J
authors.lastname: Klingler
authors.forename: Dieter
authors.initials: D
authors.lastname: Kaufmann
mesh_terms: D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D050091:Dendrimers; D005347:Fibroblasts / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D005822:Genetic Vectors; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D008113:Liver Neoplasms / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D009841:Oligonucleotides / Q000235:genetics*; D017354:Point Mutation; D011126:Polypropylenes / Q000737:chemistry*; D014162:Transfection
chemical_list: D050091:Dendrimers; D005456:Fluorescent Dyes; D009841:Oligonucleotides; D011126:Polypropylenes; C443641:poly(propyleneimine); D007041:Hypoxanthine Phosphoribosyltransferase; D016650:Fluorescein-5-isothiocyanate