Investigation intracellular trafficking of siRNAs following their delivery to cells is of great interest to elucidate dynamics of siRNA in cytoplasm. In this study, we present a novel confocal laser scanning microsco...Investigation intracellular trafficking of siRNAs following their delivery to cells is of great interest to elucidate dynamics of siRNA in cytoplasm. In this study, we present a novel confocal laser scanning microscopy (CLSM) method to evaluate a novel delivery system of 3'-peptide-siRNA therapeutic, which was named 3'-pAs-siRNA/CLD. This method could not only calculate the content of the intracellular 3'-peptide-siRNA, but also quantify its co-localization with cellular substructure. We observed that 3'-pAs-siRNA/CLD, which provided the better antitumor capability, also had a better cell uptake, endosome escape and a longer retention time in A375. This novel strategy was proved to be efficient, quantified and visualized, thus making the dynamics research of siRNA in cytoplasm clear and simplified.展开更多
基金Ministry of Science and Technology of China(Grant No.2012AA022501,2012CB720604)the National Natural Science Foundation of China(Grant No.20932001,81302626)
文摘Investigation intracellular trafficking of siRNAs following their delivery to cells is of great interest to elucidate dynamics of siRNA in cytoplasm. In this study, we present a novel confocal laser scanning microscopy (CLSM) method to evaluate a novel delivery system of 3'-peptide-siRNA therapeutic, which was named 3'-pAs-siRNA/CLD. This method could not only calculate the content of the intracellular 3'-peptide-siRNA, but also quantify its co-localization with cellular substructure. We observed that 3'-pAs-siRNA/CLD, which provided the better antitumor capability, also had a better cell uptake, endosome escape and a longer retention time in A375. This novel strategy was proved to be efficient, quantified and visualized, thus making the dynamics research of siRNA in cytoplasm clear and simplified.