XMIR-1 RNA oligo miRNA-1-3p with Xmotif

Supporting Data

XMIRs and AXMIRs can be used to modulate levels of a specific protein in target cells.

With XMIR/AXMIR technology, you can use exosomes to deliver miRNAs and anti-miRNAs to cells in order to alter levels of a specific protein (NOTE: when combined with our XStamp™ Exosome Targeting Technology, you can even generate miR/anti-miR-carrying exosomes that are targeted to a specific cell type). To demonstrate the effectiveness of using exosomes containing either miRNA or anti-miRNA to modulate protein expression levels in target cells, we generated XMIR-21 and AXMIR-21 containing exosomes from HEK293 cells, and used Western blotting to monitor expression levels of the protein PDCD4, whose expression is reduced in the presence of miR-21.

We transfected HEK293 cells with either an miR-21 XMIR Oligo (XMIR-21) or an anti-miR-21 AXMIR Oligo (AXMIR-21), isolated exosomes loaded with the respective XMIR/AXMIR, and then added these exosomes to naïve HEK-293 cells in culture. After 24 hours, total cell lysates were taken and Western blots for PDCD4, a known miR-21 target, were performed (Figure 1A), with GAPDH protein levels used as a loading control and reference signal for band intensity quantitation analysis. Quantitation is shown in Figure 1B.

These data clearly show that XMIR and AXMIR-containing exosomes can modulate expression of the target protein, as addition of exosomes from cells transfected with XMIR-21 resulted in down regulation of endogenous levels of PDCD4, and addition of exosomes from cells transfected with AXMIR-21 resulted in increased levels of PDCD4. These results also confirm that XMIRs act on endogenous targets and that AXMIRs effectively act as miRNA inhibitors in cells when delivered via exosomes.