The DGKE Knockout Raji Polyclonal Cells represent a CRISPR/Cas9-mediated gene-disrupted polyclonal population of Raji B lymphocytes, engineered for loss-of-function studies of diacylglycerol kinase epsilon (DGKE). This knockout model consists of a heterogeneous pool of edited cells, each carrying disruptions at the DGKE locus, enabling investigation of DGKE function in a B-cell context without clonal selection biases. The product provides a versatile tool for dissecting lipid signaling and B-cell receptor (BCR) pathway dynamics in human lymphoma-derived B cells.
The Raji cell line is an Epstein-Barr virus (EBV)-positive Burkitt lymphoma B lymphocyte line established from a pediatric patient. Raji cells retain key features of mature B cells, including surface immunoglobulin expression and robust BCR signaling machinery. As a model system, Raji cells are widely employed for studying BCR-mediated activation, immune surveillance mechanisms, antibody production, and oncogenic signaling. Their EBV-positive status also makes them relevant for investigating viral latency and lymphomagenesis, while their rapid proliferation facilitates high-throughput functional assays.
DGKE encodes a diacylglycerol kinase that preferentially phosphorylates diacylglycerol (DAG) to generate phosphatidic acid (PA), modulating the balance of these lipid second messengers. In BCR signaling, antigen stimulation triggers SYK and BTK, leading to PLCG2-mediated DAG production and calcium influx. DGKE attenuates DAG-driven PKC activation: by consuming DAG, DGKE limits PKC?? stimulation and subsequent CARMA1-BCL10-MALT1 complex assembly, reducing IKK-mediated NF-??B activation. PA generated by DGKE influences mTORC1 and Raf-1 signaling, integrating metabolic and survival cues. DGKE thus occupies a critical node between phospholipid signaling and transcriptional responses.
In Raji B cells, DGKE knockout removes a key negative regulator of BCR-proximal signaling. The resulting accumulation of DAG upon BCR engagement is predicted to enhance PKC?? activity, amplify calcium flux, and hyperactivate the CARMA1-BCL10-MALT1?CIKK?CNF-??B axis, leading to increased expression of activation markers such as CD69 and CD86. This dysregulated signaling may promote lymphocyte proliferation and survival, recapitulating aspects of B-cell hyperactivity and lymphomagenesis. The polyclonal nature of the knockout population ensures that diverse editing events are represented, facilitating the study of DGKE loss in a context that mirrors heterogeneous cellular responses.
This DGKE knockout model supports applications including mechanistic dissection of BCR signal transduction, lipid second messenger profiling, and diacylglycerol kinase family analysis. Researchers can perform Western blotting for PKC and I??B?? phosphorylation, flow cytometry for activation markers, calcium mobilization assays, DAG/PA lipidomics, NF-??B reporter assays, and BCR-induced proliferation or apoptosis studies. The cells are also suitable for DGK epsilon inhibitor screening and complement dysregulation modeling in atypical hemolytic uremic syndrome and thrombotic microangiopathy. RNA-sequencing can characterize DGKE-related transcriptomic changes. For further information or customization, contact Ascent Research.
