Abstract:
:The presence of about 1.2 M glycerol during transfection with DNA/transferrin-polylysine and DNA/polylysine complexes dramatically increases transgene expression in a variety of cell types, provided that the complexes have an excess of polylysine. We have characterized this phenomenon using a human melanoma cell line (H225). The addition of 1.2 M glycerol to the transfection medium has no influence on the internalization of DNA complexes or on the promoter activity used to direct reporter gene expression. Neither prenor postincubation of the cells with glycerol results in a notable increase in transgene expression. Bafilomycin A1 and chloroquine, two drugs affecting the endosomal pathway, both influenced transgene expression, indicating that glycerol acts on internal vesicles. Glycerol and polylysine synergized in their ability to lyse erythrocytes as well as internal vesicles (microsomes) isolated from H225 cells, indicating that the glycerol effect is due to a labilization of vesicular membranes, which facilitates membrane disruption by polylysine. Our current model suggests that the excess of polylysine in the DNA complexes disrupts vesicular membranes in the presence of glycerol, thus allowing the release of DNA complexes into the cytoplasm.
journal_name
Exp Cell Resjournal_title
Experimental cell researchauthors
Zauner W,Kichler A,Schmidt W,Mechtler K,Wagner Edoi
10.1006/excr.1997.3486subject
Has Abstractpub_date
1997-04-10 00:00:00pages
137-45issue
1eissn
0014-4827issn
1090-2422pii
S0014482797934868journal_volume
232pub_type
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journal_title:Experimental cell research
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