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Reagents for Up and Down-Regulation of miRNA Activity in Mammalian Cells

). Interestingly, the cells transfected with the miR-31 inhibitor become elongated and display thin membrane protrusions similar in appearance to neurite outgrowths (Figure 5, left inset). The negative control data was identical to the nontransfected control (Figure 5, right inset). Additional screening experiments using groups of Pre-miR miRNA and Anti-miR miRNA Inhibitors will undoubtedly reveal miRNAs that are involved in a broad range of cellular processes.

Figure 5. Identification of miRNAs that Alter Cell Proliferation. HeLa cells (5 x 103) were transfected with individual Anti-miR miRNA Inhibitors in triplicate using siPORT NeoFX Transfection Agent (Ambion). 72 hr post-transfection, cells were fixed and stained with propidium iodide to count total cell number (TTP LabTech Acumen Exporer). Cells were then stained for -actin using immunofluorescence.

The horizontal shaded area represents the normal range of cell number for this cell type, as exemplified by cells transfected with a negative control that does not affect cell proliferation (second bar from the right). The right inset shows the morphology of these control cells. The left inset shows the morphology of cells transfected with an Anti-miR Inhibitor to miR-31.


Pre-miR miRNA and Anti-miR miRNA Inhibitors

We have developed ready-to-use synthetic precursor miRNAs and antisense miRNAs for use in transfection experiments. These are valuable tools for functional analyses of miRNA activity in a wide variety of biological systems. See www.ambion.com/miRNA
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