The DRAM2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the DRAM2 gene in the human Raji B lymphocyte cell line. This loss-of-function model enables systematic investigation of DRAM2-dependent autophagy and apoptosis pathways in a lymphoblastoid background. The polyclonal format provides a heterogeneous pool of edited cells, avoiding clonal selection artifacts and preserving population-level responses relevant to cancer biology and drug screening.
The Raji cell line is an Epstein?CBarr virus (EBV)-positive Burkitt lymphoma model derived from a B lymphocyte lineage. These suspension cells express surface immunoglobulins and retain key features of antigen-presenting B cells, making them a widely used system for studying B-cell malignancies, immune signaling, and viral oncogenesis. Their rapid proliferation and well-characterized genomic landscape facilitate functional genomics and high-throughput loss-of-function screens, particularly for genes involved in lymphoma biology and therapy resistance.
DRAM2 (DNA damage-regulated autophagy modulator 2) encodes a lysosomal membrane protein that acts downstream of p53 and DNA damage signals to coordinate autophagy and apoptosis. Upon activation by p53 or E2F1, DRAM2 promotes autophagic flux by enhancing lysosomal acidification and autophagosome?Clysosome fusion, leading to LC3-II conversion and p62 degradation. Simultaneously, it engages the intrinsic apoptotic machinery through interactions with BAX, BAK, ATG5, and ATG7, facilitating cytochrome c release and caspase activation. This dual function positions DRAM2 at a critical node integrating p53-mediated tumor suppression, lysosomal degradation, and programmed cell death.
In the Raji lymphoma context, DRAM2 knockout disrupts the intricate balance between pro-survival autophagy and cell death, which is often subverted in B-cell malignancies. Loss of DRAM2 is expected to impair DNA damage-induced apoptosis and autophagic clearance, potentially revealing mechanisms of chemoresistance and tumor maintenance. This model is particularly relevant for dissecting p53 pathway status in EBV-driven lymphomagenesis, as EBV-encoded proteins frequently modulate both p53 and autophagy. The polyclonal knockout pool allows researchers to study these effects without the confounding influence of single-cell cloning artifacts.
This engineered cell population supports a broad range of functional studies, including Western blot analysis of autophagic markers (LC3-II, p62), flow cytometry-based apoptosis quantification via Annexin V staining, immunofluorescence microscopy for lysosomal markers (LAMP1, cathepsin D), qPCR profiling of p53 target genes, and cell viability assays under genotoxic stress. Researchers can employ this model to investigate autophagy-apoptosis crosstalk, identify novel regulators of DRAM2 signaling, screen for synthetic lethal interactions, or evaluate candidate therapeutics targeting lysosomal cell death pathways in lymphoma. For additional details on characterization and culture conditions, please contact Ascent Research.
