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Intracellular localization of lipoplexed siRNA in vascular endothelial cells of different mouse tissues.

Microvascular Research

genetics, Animals, Vascular Endothelial Growth Factor Receptor-2, metabolism, chemistry, administration & dosage, RNA, Small Interfering, cytology, Pulmonary Alveoli, PTEN Phosphohydrolase, Myocardium, Mice, Nude, Mice, Inbred C57BL, Mice, Lung, Liver, Liposomes, analysis, Laminin, Intracellular Space, Immunohistochemistry, Glycoproteins, Gene Expression, Fluorescent Antibody Technique, Endothelium, Lymphatic, Endothelial Cells, Carbocyanines, Cadherins, Antigens, CD34, Antigens, CD31, Antigens, CD

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      Abstract

      Liposomally formulated siRNA can be used for RNAi applications in vivo. Intravenous bolus administration of lipoplexed siRNA has been shown to reduce gene expression in the vascular endothelium. Here, we applied immunofluorescence staining for different endothelial markers (PECAM-1, CD34, laminin) on paraffin sections to compare the respective expression pattern with the intracellular localization of intravenously administered, fluorescently labeled siRNA (siRNA-Cy3-lipoplex). By confocal microscopy, lipoplexed siRNA-Cy3 was detected inside vascular endothelial cells in vivo, which where identified with co-staining of endothelial markers. Consequently, the finding of intracellular siRNA uptake by vascular endothelial cells correlated with RNAi based specific protein reduction in situ as revealed by PECAM-1 specific immunofluorescence staining in lung tissue sections. Therefore, by using a cell biological approach these in situ data emphasize the functional uptake of liposomal siRNA molecules in vascular endothelial cells of different mouse tissues as indicated in our previous molecular study.

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      Journal
      10.1016/j.mvr.2008.02.004
      18455200

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