This product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from Raji B lymphocytes, in which the LPIN2 gene has been disrupted using non-homologous end joining (NHEJ)-mediated repair. The resulting polyclonal pool contains a heterogeneous mix of loss-of-function mutations across the LPIN2 locus, offering a robust model for studying LPIN2-dependent processes without the clonal artifacts that may arise in single-cell-derived lines. The polyclonal format preserves the broad genetic background of the parental Raji line, enabling the analysis of LPIN2 function in the context of B cell biology while minimizing the risk of off-target clonal skewing. This knockout model supports detailed functional genomics, biochemical pathway dissection, and drug-screening applications in immunology and cancer research.
The parental Raji cell line is an Epstein-Barr virus (EBV)-positive Burkitt lymphoma-derived B lymphocyte model, widely employed to investigate B cell lymphomagenesis, EBV-driven oncogenesis, and humoral immune responses. Raji cells exhibit a mature B cell phenotype with constitutive activation of survival pathways downstream of viral latent membrane proteins, making them particularly suitable for studies linking metabolic reprogramming to malignancy. Their EBV positivity also facilitates research into virus?Chost interactions, immune evasion mechanisms, and the impact of genetic perturbations on B cell receptor signaling and cytokine production. As a well-characterized suspension cell line, Raji cells are amenable to lipidomic, proteomic, and transcriptomic analyses, providing a versatile platform for dissecting the crosstalk between lipid metabolism and oncogenic signaling.
LPIN2 encodes a magnesium-dependent phosphatidate phosphatase (PAP) enzyme that catalyzes the conversion of phosphatidic acid (PA) to diacylglycerol (DAG), a critical step in glycerolipid synthesis and signal transduction. In addition to its catalytic function, LPIN2 acts as a transcriptional coactivator, interacting with PPARGC1A, PPARG, and CEBPA to promote the expression of genes involved in adipogenesis, mitochondrial biogenesis, and fatty acid oxidation. LPIN2 is regulated by multiple upstream signals, including PPARG, SREBF1, insulin, TNF??, and mTORC1, and it modulates a network of downstream effectors such as PKC, FABP4, and the NLRP3 inflammasome. Through its dual enzymatic and coactivator roles, LPIN2 integrates nutrient and inflammatory cues to maintain lipid homeostasis and control the production of pro-inflammatory mediators like IL1B.
Disruption of LPIN2 in Raji B cells leads to the accumulation of PA and a corresponding decrease in DAG levels, impairing PKC-mediated signaling cascades that influence cell proliferation, survival, and cytokine secretion. This perturbation is expected to dysregulate phospholipid and triglyceride biosynthesis, alter the composition of lipid rafts, and affect the activation of NF-??B and inflammasome pathways. Given the role of LPIN2 variants in Majeed syndrome??a disorder characterized by recurrent fevers, osteomyelitis, and dyserythropoietic anemia??this knockout model provides a unique tool to investigate how lipid phosphatase dysfunction contributes to autoinflammation and metabolic stress in B cells. The interplay between LPIN2, PPARG, and the 14-3-3 protein scaffold further highlights its importance in coordinating metabolic and inflammatory programs in lymphoma, offering insights into potential vulnerabilities that can be exploited therapeutically.
The LPIN2 Knockout Raji Polyclonal Cells are suitable for a wide range of experimental workflows, including lipidomic profiling by LC-MS to quantify changes in PA, DAG, and glycerophospholipid species, as well as triglyceride quantification assays to assess metabolic flux. Researchers can pair this model with Western blot analysis of LPIN2 and phospho-PKC substrates, RT-qPCR for target gene expression, and NF-??B reporter assays to map signaling alterations. Additional applications include flow cytometry for apoptosis and cell cycle analysis, co-immunoprecipitation to verify LPIN2 interactions with PPARG or 14-3-3 proteins, and inflammasome activation assays to measure IL1?? release. These cells also serve as a platform for drug sensitivity profiling in B cell lymphoma and for screening compounds that target lipid-dependent oncogenic pathways. For further information, please contact Ascent Research.
