GSE169000
Gene expression profiling of MDA231 cells depleted of MRCKalpha (CDC42BPA) and MRCKbeta (CDC42BPB)
Aim: The MRCK family of kinases (namely MRCKalpha and MRCKbeta) are downstream effectors of the Rho GTPase signaling pathways with roles in mediating cell migration via regulating the actin cytoskeleon. As such, it is thought that the MRCKs might also be involved in promoting cancer cell invasion or metastasis. However, crucial in vivo data and a general view of the signaling pathways regulated by the MRCKs are still lacking in order to substantiate the hypothesis. Therefore, the goal of this study is to perform transcriptome profiling (RNA-seq) of MDA-MB-231 cells depleted of either CDC42BPA, CDC42BPB, or both, in order to obtain insight into the differential genes expressed and whether CDC42BPA and CDC42BPB contribute as a causative factor of metastatic breast cancer. Method: We performed CRIPSR-based knockout of MRCKalpha (CDC42BPA), MRCKbeta (CDC42BPB), either alone or in combination, in the invasive breast cancer cell line MDA-MB-231. The combinatory double knockout was performed twice as two biological replicates to increase confidence in our data. The knockout cells were then prepared and sent for RNA-seq using by BGISEQ-500, and differential gene expression profile was obtained. Overall design: mRNA from vector control, MRCKalpha (CDC42BPA) knockout, MRCKbeta (CDC42BPB) knockout and MRCKalpha/MRCKbeta (CDC42BPA/CDC42BPB) double knockout MDA-MB-231 cells