PMID: 9446645Feb 3, 1998Paper

Oligodeoxyribonucleotide uptake in primary human hematopoietic cells is enhanced by cationic lipids and depends on the hematopoietic cell subset.

Blood
R KronenwettR Haas

Abstract

The use of antisense oligodeoxyribonucleotides (ODN) is a potential method to switch off gene expression. The poor cellular uptake of ODN in primary cells still is a limiting factor that may contribute to the lack of functional efficacy. Various forms of cationic lipids have been developed for efficient delivery of nucleic acids into different cell types. We examined the two cationic lipids DOTAP and DOSPER to improve uptake of ODN into primary human hematopoietic cells. Using a radiolabeled 23-mer, ODN uptake into blood-derived mononuclear cells could be increased 42- to 93-fold by DOTAP and 440- to 1,025-fold by DOSPER compared with application of ODN alone. DOTAP was also effective for delivery of ODN into leukocytes within whole blood, which may resemble more closely the in vivo conditions. As assessed by fluorescein isothiocyanate-conjugated ODN both cationic lipids enhanced cytoplasmic accumulation of ODN in endosome/lysosome-like structures with a partial shift of fluorescence to the whole cytoplasm and the nucleus following an incubation of 24 hours. ODN uptake by cationic lipids into different hematopoietic cell subsets was examined by dual-color immunofluorescence analysis with subset-specific monoclonal antibodies. W...Continue Reading

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