Immunocytochemical visualization of transfected DNA in cultured cells.
Nonviral transfection is one of the modern methods for the incorporation of foreign genes into cells. This process involves uptake of foreign genetic material by the cell and further trafficking through the cytoplasm to the nucleus. Elucidation of cytoplasmic pathways of transfection complexes can be useful to improve already existing gene delivery systems or to establish new systems. To monitor transfection complexes in the cell during transfection, we elaborated a method for the visualization of transfection complexes by introducing digoxigenin-labelled nucleotides into foreign DNA followed by detection of digoxigenin label with the use of antibodies directed against digoxigenin. This procedure allowed the visualization of DNA in transfection complexes and to monitor these complexes in cells during transfection.
Zaitsev
S
S
Department of Nephrology, Franz Volhard Clinic, Max Delbrück Center for Molecular Medicine, Berlin-Buch, Germany.
Buchwalow
I
I
Haberland
A
A
Tkachuk
S
S
Böttger
M
M
D002461:Cell Line, Transformed; D004247:DNA / Q000032:analysis* / Q000235:genetics; D004076:Digoxigenin; D004730:Endothelium, Vascular / Q000737:chemistry* / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006657:Histones / Q000737:chemistry / Q000235:genetics; D006801:Humans; D007124:Immunoenzyme Techniques; D008856:Microscopy, Fluorescence; D010957:Plasmids; D014162:Transfection
D006657:Histones; D004247:DNA; D004076:Digoxigenin