The EED Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the 786-O human clear cell renal cell carcinoma (ccRCC) line, featuring targeted disruption of the EED gene. EED encodes a core scaffold subunit of the Polycomb Repressive Complex 2 (PRC2), essential for histone H3K27 trimethylation (H3K27me3) and gene silencing. As a polyclonal pool, this product enables loss-of-function studies of PRC2-dependent repression in a ccRCC context.
The parental 786-O line is a widely used model of human renal cell adenocarcinoma, derived from a primary clear cell carcinoma. It carries a homozygous VHL tumor suppressor mutation, leading to constitutive HIF pathway activation and an epithelial morphology. This genetic background creates a unique epigenetic landscape that renders the cells susceptible to perturbations of chromatin modifiers such as PRC2, making them ideal for studying epigenetic regulation in kidney cancer.
Within PRC2, EED scaffolds the catalytic subunit EZH2 and other core components SUZ12 and RBBP4/7, while interacting with accessory factors JARID2 and AEBP2. EED allosterically activates EZH2, facilitating H3K27me3 spreading that silences key genes including the tumor suppressors CDKN2A (p16INK4a) and CDH1 (E-cadherin), as well as HOX developmental regulators. Upstream signals, such as transcription factors E2F and MYC, and lncRNAs HOTAIR and Xist, can modulate PRC2 activity to influence this silencing network.
In the VHL-mutant 786-O background, EED knockout disrupts PRC2 function, leading to loss of H3K27me3 and derepression of PRC2 targets. This may reactivate tumor suppressor genes and alter cancer cell phenotypes such as proliferation and apoptosis. The model thus provides a tool to examine PRC2 dependency in ccRCC and to evaluate epigenetic therapeutics targeting this complex.
Applications include epigenetic gene regulation studies, screening of PRC2 inhibitors (e.g., EZH2 inhibitors), and investigation of drug resistance mechanisms. Common assays include Western blotting for H3K27me3, RT-qPCR and RNA-seq for target gene expression, ChIP-seq for histone modifications, cell proliferation and colony formation assays, and drug sensitivity testing. These cells also support differentiation induction and tumor suppressor reactivation research. For further information, contact Ascent Research.
