The DDHD2 Knockout Raji Polyclonal Cells are a heterogeneous population of Raji B lymphocytes engineered via CRISPR/Cas9-mediated gene disruption to ablate DDHD2 expression. This polyclonal knockout pool provides a versatile loss-of-function model for interrogating the biological roles of the intracellular phospholipase A1 DDHD2 in a human B-cell lymphoma background. Unlike monoclonal knockout cell lines, the polyclonal format captures a spectrum of editing events, reflecting the inherent genetic diversity within the population and enabling robust functional studies without the clonal selection artifacts that may arise from single-cell expansion.
The Raji host cell line is an EBV-positive human B lymphocyte line from a Burkitt lymphoma patient. Widely used in immunology and cancer research, Raji cells model B-cell receptor signaling, lymphomagenesis, and apoptosis. Their rapid suspension growth and characterized genome make them ideal for gene function studies in B-cell malignancies.
DDHD2 encodes an intracellular phospholipase A1 that hydrolyzes phosphatidylcholine (PC) and phosphatidylethanolamine (PE) to produce lysophosphatidylcholine (LPC) and lysophosphatidylethanolamine (LPE), releasing free fatty acids. These lipid metabolites are critical for membrane remodeling, autophagosome biogenesis, and mitochondrial lipid homeostasis. DDHD2 activity is functionally linked to autophagy, promoting LC3 lipidation and p62 degradation, and its loss results in lipid droplet accumulation and impaired autophagy flux. The enzyme operates at endoplasmic reticulum and mitochondrial membrane contact sites, likely coordinating with autophagic machinery. DDHD2 functions downstream of unknown regulators, possibly responsive to cellular lipid status.
In the Raji B-cell lymphoma context, DDHD2 knockout disrupts the delicate balance of phospholipid metabolism, which may have pleiotropic effects on cell proliferation, survival, and stress responses. B-cell lymphomas often exhibit altered lipid metabolism to support rapid growth and evade apoptosis, making them particularly sensitive to perturbations in lipid-modifying enzymes. By eliminating DDHD2, this model enables dissection of how phospholipid turnover influences autophagy-dependent survival mechanisms, mitochondrial fitness, and lipid second messenger generation in malignant B cells. The polyclonal nature of the knockout population mimics the genetic heterogeneity found in tumors, offering a more representative system for drug response studies and functional genomics than clonal lines.
This knockout model supports diverse applications: Western blot and RT-qPCR confirm DDHD2 ablation; lipidomic profiling (LC-MS) quantifies changes in LPC and LPE; autophagy flux assays with chloroquine distinguish autophagosome accumulation from flux defects. Flow cytometry measures apoptosis (annexin V/PI) and proliferation; MitoTracker staining assesses mitochondrial membrane potential; immunofluorescence visualizes lipid droplets and LC3 puncta. The cells are also suitable for drug screening to identify modulators of lipid metabolism and autophagy, and for modeling SPG54, a hereditary spastic paraplegia linked to DDHD2 mutations. DDHD2 Knockout Raji Polyclonal Cells thus facilitate translational research in oncology and neurology. For technical inquiries, please contact Ascent Research.
