EED Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from A-549 human lung adenocarcinoma epithelial cells, featuring targeted disruption of the EED gene. This loss-of-function model enables investigation of PRC2 complex function in non-small cell lung cancer (NSCLC) without the bias of clonal selection.
The A-549 host cell line, established from a 58-year-old Caucasian male with lung adenocarcinoma, displays adherent epithelial morphology and serves as a widely used NSCLC model. A-549 cells retain type II alveolar epithelial characteristics and harbor KRAS mutation, making them relevant for studying epigenetic dependencies in RAS-driven tumors.
EED encodes a core PRC2 subunit that allosterically activates EZH2 methyltransferase to propagate H3K27me3 repressive marks, silencing genes such as HOX clusters, CDKN2A (p16INK4a), and CDH1 (E-cadherin). EED interacts with EZH2, SUZ12, RBBP4/RBBP7, and accessory factors JARID2 and AEBP2. Its expression is regulated by MYC and E2F in a cell cycle-dependent manner, connecting PRC2 activity to proliferation.
Disruption of EED in A-549 cells destabilizes PRC2, reduces H3K27me3, and reactivates silenced tumor suppressors, providing a model to dissect roles of epigenetic silencing in lung adenocarcinoma maintenance, metastasis, and cancer stem cell biology. This knockout system also supports evaluation of EED-targeted inhibitors and exploration of Weaver syndrome-associated mutations.
These polyclonal knockout cells are suited for Western blot analysis of H3K27me3, ChIP-qPCR at target loci, RT-qPCR of derepressed genes, and functional assays including proliferation, viability, migration, and invasion. Drug sensitivity testing with EED inhibitors (e.g., EED226, MAK683) and RNA-seq transcriptomic profiling are readily performed. Flow cytometry enables cell cycle analysis, and colony formation assays assess clonogenic potential. For further information, please contact Ascent Research.
