Glycosylated polylysine/DNA complexes: gene transfer efficiency in relation with the size and the sugar substitution level of glycosylated polylysines and with the plasmid size.
A DNA delivery system based on the use of polylysine substituted with small recognition signals, such as carbohydrate moieties specifically recognized by membrane lectins present in a given cell line, has been developed [Midoux et al. (1993) Nucleic Acids Res. 21, 871-878]. Human hepatoma (HepG2) cells which express a galactose-specific membrane lectin are efficiently transfected in the presence of chloroquine with pSV2Luc plasmid complexed with a lactosylated polylysine. The optimization of the parameters involved in the formation of DNA/glycosylated polylysine complexes leads to the following conclusions: a high gene transfer efficiency is reached when (i) DNA/glycosylated polylysine complexes are completely retarded when subjected to electrophoresis and when (ii) 31 +/- 4% or 40 +/- 8% of the amino groups of a polylysine having a degree of polymerization (DP) of 190 are substituted with lactosyl or beta-D-galactosyl residues, respectively. In addition, carbohydrate residues bound to polylysine decrease the electrostatic strength between plasmid DNA and glycosylated polylysine, suggesting that the strength of the electrostatic interactions between the plasmid and the glycosylated polylysine plays an important role in the efficiency of the gene expression. The optimal lactosylated polylysine conjugate (polylysine DP 190 substituted with 60 lactosyl residues) transfers a 5 kb and a 12 kb plasmid with a similar efficiency.
Erbacher
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P
Centre de Biophysique Moléculaire BatB, CNRS et Université d'Orléans, France.
Roche
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AC
Monsigny
M
M
Midoux
P
P
D006528:Carcinoma, Hepatocellular; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002738:Chloroquine / Q000494:pharmacology; D004337:Drug Carriers / Q000138:chemical synthesis; D005690:Galactose; D006031:Glycosylation; D006801:Humans; D007700:Kinetics; D037102:Lectins / Q000378:metabolism; D008113:Liver Neoplasms; D008156:Luciferases / Q000032:analysis / Q000096:biosynthesis; D009994:Osmolar Concentration; D011107:Polylysine; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis*; D013329:Structure-Activity Relationship; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured
D004337:Drug Carriers; D037102:Lectins; D011994:Recombinant Proteins; D011107:Polylysine; D002738:Chloroquine; D008156:Luciferases; D005690:Galactose