effect of RNA uptake on gene expressions in human NK and CD8-positive T cells

In the blood, non-vesicular extracellular RNA functions as a cell-cell communication molecule. We have identified that IL1b-mRNA fragment binds ZC3H12D protein on the cell surface. This protein expresses on NK and T cells. When extracellular RNA binds ZC3H12D on the cell surface, the RNA-ZC3H12D complex goes inside the cell to modulate cell character, and as a result, the RNA stimulated-immune cell became anti-tumor rather than pro-tumor. To decipher the molecular mechanisms to govern the extracellular RNA-mediated signaling, we carried out NGS analysis to find out the difference in gene expression profiles after external RNA stimulation. Overall design: Human PBMC was purchased from Roche. Thawed PBMC was maintained in HPLM (Thermo) medium supplemented with 10% FCS. The following day, FITC labeled IL1b-mRNA (10ng/mL) was applied in the media, and 0.5h or 6h later cells were FACS-sorted to isolate NK cells (CD56+CD3-) and CD8+T cells. Isolated cells were immediately placed into Trizol (Thermo) reagent to obtain total RNA.