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Evidence for the Role of Proteoglycans in Cation-Mediated Gene Transfer
Kimberly A. Mislick and John D. Baldeschwieler
Proceedings of the National Academy of Sciences of the United States of America
Vol. 93, No. 22 (Oct. 29, 1996), pp. 12349-12354
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/40620
Page Count: 6
You can always find the topics here!Topics: HeLa cells, Transfection, CHO cells, Proteoglycans, Cell lines, Sulfates, Chlorates, Cells, Glycosaminoglycans, Lipids
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We report evidence that gene complexes, consisting of polycations and plasmid DNA enter cells via binding to membrane-associated proteoglycans. Treatment of HeLa cells with sodium chlorate, a potent inhibitor of proteoglycan sulfation, reduced luciferase expression by 69%. Cellular treatment with heparinase and chondroitinase ABC inhibited expression by 78% and 20% with respect to control cells. Transfection was dramatically inhibited by heparin and heparan sulfate and to a smaller extent by chondroitan sulfate B. Transfection of mutant, proteoglycan deficient Chinese hamster ovary cells was 53$\times $ lower than of wild-type cells. For each of these assays, the intracellular uptake of DNA at 37 degrees C and the binding of DNA to the cell membrane at 4 degrees C was impaired. Preliminary transfection experiments conducted in mutant and wild-type Chinese hamster ovary cells suggest that transfection by some cationic lipids is also proteoglycan dependent. The variable distribution of proteoglycans among tissues may explain why some cell types are more susceptible to transfection than others.
Proceedings of the National Academy of Sciences of the United States of America © 1996 National Academy of Sciences