Cytokine Stimulation of ADAR-deleted B16 murine melanoma cells

ADAR was deleted from the B16 murine melanoma line using CRISPR/Cas9. The loss-of-function phenotype was assessed in vitro with cytokine stimulation or vehicle control. In order to determine the effect of an interferon sensitivity phenotype caused by ADAR deletion, B16 murine melanoma cancer cells were transiently transfected with plasmids encording Cas9 and guide RNA (sgRNA) against Adar or a non-targeting sequence. Cells were then subcloned and assessed for Adar deletion. Once verified, the Adar-null cells were treated with vehicle control (PBS with 0.1% BSA), interferon-beta (IFNb), or interferon-gamma (IFNg), and transcriptionally profiled by directional, paired-end RNAseq after a 36 hour stimulation.