The LPCAT2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Raji B-lymphocyte line, characterized by targeted disruption of the LPCAT2 gene. This loss-of-function model preserves a polyclonal genetic background, avoiding the clonal artifacts that can arise from single-cell isolation, and is designed for robust investigation of LPCAT2-dependent processes in a malignant B-cell context.
The Raji cell line originates from an Epstein-Barr virus (EBV)-positive Burkitt??s lymphoma and serves as a widely used model for B-cell malignancies. These cells exhibit constitutive nuclear factor-kappa B (NF-??B) activation due to EBV latent gene expression, which primes them for rapid proliferation and provides a relevant oncogenic background. Raji cells are amenable to genetic editing and retain key signaling pathways, making them an ideal host for dissecting gene function in lymphoma biology.
LPCAT2 encodes an acyltransferase that catalyzes the acetylation of lyso-platelet-activating factor (lyso-PAF) to produce platelet-activating factor (PAF), using acetyl-CoA as a co-substrate. LPCAT2 expression is transcriptionally regulated by NF-??B, acting downstream of Toll-like receptor 4 (TLR4) activation by lipopolysaccharide (LPS) or tumor necrosis factor-alpha (TNF-??). The generated PAF is a bioactive phospholipid that signals through its G-protein-coupled receptor (PAFR) to mobilize intracellular calcium and trigger cytokine release. LPCAT2 also participates in phosphatidylcholine remodeling and interacts with PAF acetylhydrolase, which degrades PAF, and other acyltransferases. In the EBV-driven Raji background, elevated NF-??B activity likely enhances LPCAT2 expression, linking viral oncogenesis to pro-inflammatory lipid mediator production.
Disrupting LPCAT2 in Raji cells abolishes de novo PAF synthesis, providing a unique platform to examine how loss of this lipid mediator alters membrane lipid composition and influences downstream signaling via the PAFR?Ccalcium?CNF-??B axis. The model is particularly valuable for studying the role of LPCAT2-dependent phospholipid remodeling in B-cell proliferation, apoptosis resistance, and cytokine milieu. By knocking out LPCAT2, researchers can interrogate compensatory pathways, such as alternative acyltransferase activities, and assess the contribution of endogenous PAF to Burkitt??s lymphoma pathophysiology.
This polyclonal knockout product is well-suited for a broad range of assays, including LC-MS-based lipidomics to profile phospholipid changes, PAF-specific enzymatic activity measurements, western blotting for pathway component expression, RT-qPCR for transcript analysis, and flow cytometry to assess calcium mobilization or surface receptor expression. Cell proliferation (MTT), apoptosis, and cytokine secretion (ELISA) assays can be employed to link LPCAT2 function to cellular outcomes. The cells enable mechanistic studies of the TLR4?CNF-??B?CLPCAT2?CPAF signaling cascade in the contexts of inflammation, asthma, thrombosis, and cancer, and support drug screening campaigns targeting PAF biosynthesis or PAFR signaling. For more information, please contact Ascent Research.
