Reactivation of SALL2 in Human Ovarian Cancer Cells
Abstract: Ovarian cancer is the seventh-most common cancer among women and the eighth-most common cause of death from cancer, and it resulted in 161,100 deaths in 2015 alone. p150 is a tumor suppressor transcription factor that has proapoptotic and growth arrest properties, essential for cells to enter a quiescent state. The p150 protein is encoded by Spalt-like gene-2 (SALL2) and studies suggested that p150 is absent or reduced in over 90% of ovarian cancers, largely due to SALL2 promoter hypermethylation. Our current study focuses on targeted demethylation of the SALL2 promoter region using a modified CRISPR based demethylase system to possibly reactivate SALL2 in ovarian cancer cells. Specific sites on the promoter region were targeted by dCas9-demethylase using small guide RNA clones. The re-expression of p150 and the methylated state of the promoter region are to be analyzed by western blot and pyrosequencing respectively. Preliminary western blot analysis of transfected ovarian carcinoma cells showed reactivation of SALL2 with demethylase and guide RNA clones, indicating the feasibility of our promoter demethylation approaches.
Jeffrey Schoen* and Swetha Vempati
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