Coupling of adenovirus to transferrin-polylysine/DNA complexes greatly enhances receptor-mediated gene delivery and expression of transfected genes.
We are developing efficient methods for gene transfer into tissue culture cells. We have previously shown that coupling of a chimeric adenovirus with polylysine allowed the construction of an adenovirus-polylysine-reporter-gene complex that transferred the transporter gene with great efficiency into HeLa cells. We have now explored simpler, biochemical means for coupling adenovirus to DNA/polylysine complexes and show that such complexes yield virtually 100% transfection in tissue culture cell lines. In these methods adenovirus is coupled to polylysine, either enzymatically through the action of transglutaminase or biochemically by biotinylating adenovirus and streptavidinylating the polylysine moiety. Combination complexes containing DNA, adenovirus-polylysine, and transferrin-polylysine have the capacity to transfer the reporter gene into adenovirus-receptor- and/or transferrin-receptor-rich cells.
Wagner
E
E
Research Institute of Molecular Pathology, Vienna, Austria.
Zatloukal
K
K
Cotten
M
M
Kirlappos
H
H
Mechtler
K
K
Curiel
D T
DT
Birnstiel
M L
ML
D000260:Adenoviruses, Human / Q000235:genetics*; D000818:Animals; D002478:Cells, Cultured; D002738:Chloroquine / Q000494:pharmacology; D004247:DNA / Q000008:administration & dosage*; D004705:Endocytosis; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D066298:In Vitro Techniques; D008099:Liver / Q000166:cytology; D051379:Mice; D011107:Polylysine / Q000737:chemistry*; D011990:Receptors, Transferrin / Q000502:physiology; D011991:Receptors, Virus / Q000502:physiology; D014162:Transfection; D014168:Transferrin / Q000378:metabolism*
D011990:Receptors, Transferrin; D011991:Receptors, Virus; D014168:Transferrin; D011107:Polylysine; D002738:Chloroquine; D004247:DNA