Influence of DNA delivery method on gene targeting frequencies in human cells.
Gene targeting can be used for genetic studies of human cell lines and has significant potential for somatic cell gene therapy. These applications are however restricted by the low frequency of homologous recombination in higher eukaryotes compared to the relatively efficient nonhomologous integration of transfected DNA into the genome. As part of our attempts to overcome this problem, we compared two widely used transfection methods for their efficiency in gene targeting. To our surprise we found that, for conditions that render similar frequencies of nonhomologous integrants, lipofection is much less efficient than electroporation in generating targeted clones. This suggests that nonhomologous and homologous recombination have different requirements for DNA delivery in human cells.
MRC Clinical Sciences Centre, Imperial College School of Medicine, Hammersmith Hospital, London, United Kingdom.
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RJ
Yáñez
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Porter
D002460:Cell Line; D004274:DNA, Recombinant / Q000008:administration & dosage* / Q000235:genetics; D018274:Electroporation; D005069:Evaluation Studies as Topic; D018390:Gene Targeting / Q000379:methods*; D015316:Genetic Therapy; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D014162:Transfection
D004274:DNA, Recombinant; D007041:Hypoxanthine Phosphoribosyltransferase