DNA condensation for gene therapy as monitored by atomic force microscopy.
The atomic force microscope (AFM) was used to assay the extent of DNA condensation in approximately 100 different complexes of DNA with polylysine (PL) or PL covalently attached to the glycoproteins asialoorosomucoid (AsOR) or orosomucoid (OR). The best condensation of DNA was obtained with 10 kDa PL covalently attached to AsOR, at a lysine:nucleotide (Lys:nt) ratio of 5:1 or higher. These conditions produce large numbers of toroids and short rods with contour lengths of 300-400 nm. Some DNA condensation into shortened thickened structures was seen with 10 kDa PL attached to AsOR at Lys:nt ratios of 1.6:1 and 3:1. Some DNA condensation was also seen with 4 kDa PL at Lys:nt ratios of 3:1 and higher. Little DNA condensation was seen with PL alone or with PL convalently attached to OR at Lys:nt ratios up to 6:1. AsOR-PL enhanced gene expression in the mouse liver approximately 10- to 50-fold as compared with PL alone.
Hansma
H G
HG
Department of Physics, University of California, Santa Barbara, CA 93106, USA. hhansma@physics.ucsb.edu
Golan
R
R
Hsieh
W
W
Lollo
C P
CP
Mullen-Ley
P
P
Kwoh
D
D
D000818:Animals; D001212:Asialoglycoproteins; D004247:DNA / Q000235:genetics* / Q000648:ultrastructure*; D018014:Gene Transfer Techniques; D008099:Liver / Q000378:metabolism; D008156:Luciferases / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D018625:Microscopy, Atomic Force / Q000379:methods*; D009961:Orosomucoid / Q000031:analogs & derivatives; D010957:Plasmids; D011107:Polylysine
D001212:Asialoglycoproteins; D009961:Orosomucoid; C017030:asialoorosomucoid; D011107:Polylysine; D004247:DNA; D008156:Luciferases