Single-stranded DNA condensed with poly-L-lysine results in nanometric particles that are significantly smaller, more stable in physiological ionic strength fluids and afford higher efficiency of gene delivery than their double-stranded counterparts.
Nonviral gene transfer vectors have been actively studied in the past years in order to obtain structural entities with minimum size and defined shape. The final size of a gene transfer vector, which is compacted into unimolecular complexes, is directly proportional to the mass of the nucleic acid to be compacted. Thus, the purpose of this study was to assess the possibility of producing ssDNA vectors and their biophysical and biological characterization. We have obtained ssDNA/poly-L-lysine complexes that are significantly smaller than their double-stranded counterparts. We have also identified a lesser aggregative behavior of compacted single-stranded vs. double-stranded DNA vectors in the presence of physiological NaCl concentrations. Expression of compacted ssDNA is observed in hepatoma cell lines. Moreover, we have successfully delivered galactosylated ssDNA complexes into cells that express the asialoglycoprotein receptor via receptor-mediated endocytosis. The reduced size and biophysical behavior of ssDNA vectors may provide an advantage for transfection of eukaryotic cells.
Unitat de Bioquímica, Departament de Ciències Fisiològiques II, Universitat de Barcelona, Feixa Llarga s/n, 08907 Hospitalet, Spain.
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Perales
D004277:DNA, Single-Stranded / Q000737:chemistry*; D005434:Flow Cytometry; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D008854:Microscopy, Electron; D009691:Nucleic Acid Denaturation; D009994:Osmolar Concentration; D010316:Particle Size; D011107:Polylysine / Q000737:chemistry*; D012996:Solutions; D014162:Transfection
D004277:DNA, Single-Stranded; D012996:Solutions; D011107:Polylysine