Endocytic uptake of fluorescence labelled DNA in yeast.
After dispiriting results using viral vectors in gene therapy, by which a number of patients acquired cancer as a result of the use of retroviral vector constructs, the percentage of non-viral approaches has increased over recent years. To elucidate potential bottlenecks in the non-viral transfection process we here introduce a novel method to directly visualize endocytic non-viral DNA uptake in a transfection approach. This novel method allows for the first time to monitor the location of DNA which is taken up by endocytosis in yeast (Saccharomyces cerevisiae) wild type and mutant strains. More specifically it enables drawing conclusions about conditions favouring non-viral gene transfection.
Riechers
Sean-Patrick
SP
Institute for Biotechnology, Department Microbiology and Genetics, Technische Universität Berlin, Berlin, Germany.
Stahl
Ulf
U
Lang
Christine
C
D004247:DNA / Q000378:metabolism*; D015342:DNA Probes / Q000737:chemistry; D004705:Endocytosis; D004996:Ethidium / Q000737:chemistry; D019793:Fluorescein / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D005822:Genetic Vectors / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
D015342:DNA Probes; D005456:Fluorescent Dyes; D004247:DNA; D004996:Ethidium; D019793:Fluorescein