Bulk RNA-seq after Overexpression of EFNB2 in U266 Multiple Myeloma Cells
EFNB2 is a cell transmembrane protein required for the survival of multiple myeloma cells. Specifically, we have previously shown that EFNB2 reverse signaling is required for myeloma proliferation and survival. Overexpressing EFNB2 increases myeloma survival, proliferation, and resistance to treatment. This effect is mediated partially through STAT5, but other mediators are likely invovled. To determine if other mediators and pathways might be involved, we performed RNA-seq on myeloma cells overepxressing EFNB2. Overall design: U266 cells were transduced using a lentivirus to express dCas9-VPR. They were then transduced to express either control (non-targeting, scrambled) guide RNA (cgRNA) or guide RNA targeting the start site of EFNB2 ("gRNA2). There are 5 replicates of controls and 5 replicates of EFNB2 gRNA. We validated that the gRNA to EFNB2 causes a 80 - 100 fold increase in EFNB2 RNA by qPCR.