XMIR-34a RNA oligo miRNA-34a-5p with Xmotif

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.

Figure 1. XMIR-21 knockdown and AXMIR-21 upregulation of the miR-21 target PDCD4. (A) Western blot data and (B) quantitation.