The EZH2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from Raji B lymphocytes, featuring disruption of the EZH2 gene. This product provides a loss-of-function model to study EZH2-dependent epigenetic regulation. The polyclonal nature ensures a heterogeneous knockout pool suitable for bulk functional assays.
Raji cells are an Epstein-Barr virus (EBV)-positive B lymphoblastoid cell line established from a Burkitt lymphoma patient. They serve as a widely used model for B-cell malignancies, characterized by rapid proliferation and retention of key B-cell signaling pathways. The EBV positivity and Burkitt lymphoma origin make them particularly relevant for studying oncogenic mechanisms in lymphoid cancers.
EZH2 encodes the catalytic subunit of Polycomb repressive complex 2 (PRC2), which trimethylates histone H3 at lysine 27 (H3K27me3), leading to chromatin compaction and transcriptional silencing. EZH2 activity is regulated by upstream signals mediated by E2F transcription factors, MYC, and the AKT and MAPK/ERK pathways. Within PRC2, it interacts directly with SUZ12, EED, RBBP4, and RBBP7, which are essential for complex assembly and histone methyltransferase function. EZH2-mediated H3K27me3 represses critical tumor suppressors including CDKN1A (p21), CDKN2A (INK4A/ARF), and CDH1 (E-cadherin), and modulates HOX gene clusters and DAB2IP. In B lymphocytes, EZH2 also participates in signal-dependent transcriptional programs downstream of B cell receptor, NF-??B, and Notch signaling, linking extrinsic cues to epigenetic silencing.
In Raji cells, which exhibit an activated B-cell phenotype and EBV-driven proliferation, EZH2 is frequently overexpressed, contributing to lymphomagenesis by enforcing H3K27me3-mediated repression of growth-suppressive genes. Disruption of EZH2 in this polyclonal knockout population abolishes PRC2 catalytic activity, leading to global reduction of H3K27me3 and transcriptional derepression of target loci. This enables investigation of EZH2??s role in maintaining the oncogenic state of Burkitt lymphoma and other B-cell malignancies, and its impact on epigenetic plasticity and therapeutic response.
Typical applications include chromatin immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) to assess H3K27me3 deposition at specific promoters, Western blotting for EZH2 and H3K27me3 levels, RT-qPCR and RNA sequencing to profile global transcriptional changes, and flow cytometry to monitor cell cycle distribution and apoptosis. The model is suitable for drug sensitivity testing with EZH2 inhibitors such as Tazemetostat and for co-immunoprecipitation experiments examining PRC2 complex integrity and composition. For further information, please contact Ascent Research.
