DGKZ Knockout Raji Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population derived from the human B lymphoblast Raji cell line, featuring targeted disruption of the DGKZ gene. These cells provide a robust loss-of-function model to interrogate DGKZ-dependent signaling mechanisms in a B-lymphoid context. The polyclonal knockout format ensures a genetically heterogeneous population, capturing a range of functional ablation effects while avoiding clonal selection artifacts. This product is designed for advanced research applications requiring stable, long-term interrogation of diacylglycerol metabolism and its pleiotropic impacts on immune cell biology.
Raji cells are an EBV-positive, suspension-growing human B lymphoblast line originally isolated from a Burkitt lymphoma patient. This cell line retains key features of mature B lymphocytes, including surface immunoglobulin expression, antigen-presenting capacity, and robust B cell receptor (BCR) signaling machinery. Raji cells serve as a widely accepted model for B cell malignancies and have been instrumental in dissecting signal transduction networks governing lymphocyte activation, proliferation, and survival. Their genetic tractability and well-characterized signaling make them an ideal host for CRISPR-based gene disruption studies.
DGKZ encodes a diacylglycerol kinase that catalyzes the conversion of diacylglycerol (DAG) to phosphatidic acid (PA), thereby terminating DAG-mediated signaling. In lymphocytes, DGKZ functions downstream of antigen receptor stimulation: upon BCR or TCR engagement, upstream kinases (e.g., Src family and SYK/ZAP70) activate phospholipase C??1 to generate DAG, which recruits PKC isoforms and RasGRP proteins. DGKZ metabolizes this DAG pool, restricting activation of PKC?? and RasGRP1, limiting downstream MAPK (ERK), mTOR, and NF-??B pathways. DGKZ also interacts with SNX27, Dlg1, 14-3-3 proteins, and F-actin, suggesting scaffolding roles. Knockout disrupts this negative regulation, leading to sustained DAG accumulation and enhanced signal output.
In Raji B cells, DGKZ knockout likely potentiates BCR-derived signals, amplifying PKC/RasGRP1-dependent transcriptional programs involved in activation and proliferation. This model enables dissection of how dysregulated DAG/PA balance influences lymphoma cell biology, given that aberrant BCR signaling and NF-??B activity are hallmarks of many B cell malignancies. The EBV-positive background further permits investigation of viral-host interactions on metabolic and signaling checkpoints. Additionally, because DGKZ has been implicated in T cell anergy, this system offers a comparative platform to study conserved and divergent roles of DGKZ across lymphocyte lineages.
Typical applications include quantitative analysis of BCR-induced phospho-signaling by Western blotting for phospho-ERK and PKC substrates, flow cytometric assessment of activation markers (CD69, CD86), and calcium flux assays. Researchers can perform lipidomics for DAG/PA species, co-immunoprecipitation to map interactomes (e.g., SNX27, RasGRP1), and RT-qPCR for downstream effectors such as NFAT and AP-1. The cells are amenable to proliferation and apoptosis studies, as well as drug screening for DGKZ inhibitors. By enabling manipulation of the DAG/PA rheostat, this knockout model supports mechanistic studies in immunology, cancer biology, and therapeutic development. For further details, please contact Ascent Research.
