Cellular Internalization of Modified miRNAs

Example 2

Once miRNAs were modified by elongation and fluorescently marked to enable intracellular tracking of modified miRNAs, Applicants assessed cellular internalization of PS-modified miRNAs by flow cytometry including PO-modified miRNA as negative non-internalizing controls. Human multiple myeloma cells MM.1 S were incubated either for 30 min or for 48 hrs with modified miRNA as indicated and analyzed by flow cytometry to assess cellular load of cells with modified miRNA. For modified let7a-3p miRNA (FIGS. 2A and 2B) and modified let7a-5p miRNA (FIGS. 4A and 4B), 10 μg/ml was used for both 30 min and 48 hr incubation. For miR17-3p miRNA (FIGS. 6A and 6B), modified miR17-5p miRNA (FIGS. 8A and 8B) and modified miR218-5p miRNA (FIGS. 10A and 10B), 20 μg/ml was used for 30 min incubation and 10 μg/ml was used for 48 hr incubation, respectively.