Nanoscopic structure of DNA condensed for gene delivery.
Scanning force microscopy was used to examine DNA condensates prepared with varying stoichiometries of lipospermine or polyethylenimine in physiological solution. For the first time, individual DNA strands were clearly visualized in incomplete condensates without drying. Using lipospermine at sub-saturating concentrations, discrete nuclei of condensation were observed often surrounded by folded loops of DNA. Similar packing of DNA loops occurred for polyethylenimine-induced condensation. Increasing the amount of the condensing agent led to the progressive coalescence or aggregation of initial condensation nuclei through folding rather than winding the DNA. At over-saturating charge ratios of the cationic lipid or polymer to DNA, condensates had sizes smaller than or equal to those measured previously in electron micrographs. Polyethylenimine condensates were more compact than lipospermine condensates and both produced more homogeneously compacted plasmids when used in a 2-4-fold charge excess. The size and morphology of the condensates may affect their efficiency in transfection.
Dunlap
D D
DD
DIBIT, San Raffaele Scientific Institute, via Olgettina 58, 20132 Milan, Italy. dunlapd@dibit.hsr.it
Maggi
A
A
Soria
M R
MR
Monaco
L
L
D000538:Aluminum Silicates / Q000737:chemistry; D004247:DNA / Q000737:chemistry / Q000648:ultrastructure*; D018014:Gene Transfer Techniques; D005998:Glycine / Q000031:analogs & derivatives / Q000737:chemistry; D018625:Microscopy, Atomic Force; D010455:Peptides / Q000737:chemistry; D010957:Plasmids; D011094:Polyethyleneimine / Q000737:chemistry; D013096:Spermine / Q000031:analogs & derivatives / Q000737:chemistry
D000538:Aluminum Silicates; D010455:Peptides; C061134:dioctadecylamidoglycylspermine; C008973:polyornithine; D013096:Spermine; D011094:Polyethyleneimine; D004247:DNA; D005998:Glycine; C011934:mica