EED Knockout MES-OV Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human ovarian endometrioid adenocarcinoma cell line MES-OV. This heterogeneous pool carries disruptions in the EED gene, encoding a core subunit of Polycomb Repressive Complex 2 (PRC2). The CRISPR/Cas9-mediated gene disruption abolishes EED expression, impairing PRC2 integrity and function, and provides a robust model for studying gene silencing. As a polyclonal population, these cells avoid clonal artifacts while retaining genetic diversity, making them suitable for a wide range of molecular and cellular assays in an ovarian cancer context.
The parental MES-OV cell line is an adherent epithelial line established from an ovarian endometrioid adenocarcinoma, a subtype with distinct epigenetic dysregulation. MES-OV serves as a clinically relevant model for examining the roles of chromatin modifiers in ovarian tumorigenesis, particularly because components of PRC2 are frequently altered in this cancer type. The cells’ adherent growth characteristics facilitate standard culture conditions and make them amenable to downstream analyses such as chromatin immunoprecipitation and transcriptomic profiling, enabling precise investigation of EED-dependent epigenetic control.
EED functions as an essential scaffold protein within PRC2, binding to the trimethylated lysine 27 of histone H3 (H3K27me3) and allosterically activating the methyltransferase activity of EZH2. This allosteric regulation propagates the H3K27me3 repressive mark and maintains gene silencing at target loci. EED directly interacts with EZH2, SUZ12, RBBP4/7, and accessory factors including JARID2 and AEBP2 to form the holocomplex. Upstream, PRC2 activity is modulated by H3K27me3 itself and signaling pathways such as AKT. Downstream, PRC2-mediated H3K27me3 deposition silences tumor suppressor genes, including CDKN2A, PTEN, and APC, as well as developmental regulators. Knockout of EED disrupts the allosteric feedback loop, resulting in global loss of H3K27me3 and derepression of these targets, leading to transcriptional reprogramming that affects cell proliferation and survival.
In ovarian cancer, aberrant H3K27me3 patterns silence tumor suppressors, promoting oncogenesis. EED Knockout MES-OV Polyclonal Cells allow dissection of PRC2’s role in malignancy within the endometrioid context. Comparing knockout and parental cells reveals direct targets and compensation by other modifiers. Because EED allosterically activates PRC2, these cells are ideal for testing compounds targeting the EED?CH3K27me3 interaction, distinct from EZH2 catalytic inhibitors.
These polyclonal knockout cells are suited for epigenetic and oncology research. Applications include Western blotting for EED and H3K27me3 levels, ChIP-qPCR for H3K27me3 occupancy, RT-qPCR for target derepression (e.g., CDKN2A, PTEN), RNA-seq for transcriptome changes, and flow cytometry for cell cycle and apoptosis. In drug discovery, they enable PRC2 inhibitor screening and target engagement studies. For further information, please contact Ascent Research.
