The FDFT1 Knockout Raji Polyclonal Cells product provides a heterogeneous, CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B lymphocyte line, featuring targeted disruption of the FDFT1 gene. This ready-to-use polyclonal population bypasses clonal selection and enables immediate functional investigation of squalene synthase in a lymphoma background.
Raji cells are an Epstein-Barr virus (EBV)-positive Burkitt lymphoma-derived lymphoblastoid B-cell line extensively utilized as a model for B-cell malignancies, immune synapse studies, and antibody-dependent cellular cytotoxicity. Their robust proliferation and well-characterized signaling pathways render them a suitable host for metabolic pathway interrogation.
FDFT1 encodes squalene synthase, an NADPH-dependent enzyme that catalyzes the dimerization of farnesyl diphosphate to squalene, committing carbon flux to sterol biosynthesis. Transcriptional activation of FDFT1 is driven by sterol regulatory element-binding proteins SREBP1 and SREBP2 under low-sterol conditions, while cholesterol and oxysterols promote SREBP retention in the endoplasmic reticulum through INSIG proteins and SCAP. The squalene generated is subsequently converted to sterols by downstream enzymes including squalene epoxidase (SQLE) and lanosterol synthase (LSS), linking the mevalonate pathway to cholesterol homeostasis and isoprenoid synthesis.
In the Raji lymphoma model, knockout of FDFT1 abrogates squalene production, thereby disrupting de novo cholesterol synthesis and potentially compromising membrane lipid raft organization and SREBP-mediated lipid homeostasis. These alterations can influence B-cell receptor signaling, cytokine responses, and cell survival, making the polyclonal knockout population a powerful system to study the dependence of B-cell malignancies on the mevalonate pathway and to identify metabolic vulnerabilities. This model enables researchers to explore how cholesterol metabolism supports lymphoma growth and to test small-molecule inhibitors targeting the mevalonate pathway.
The FDFT1 Knockout Raji Polyclonal Cells are suited for a range of experimental applications, including cholesterol biosynthesis studies, drug target validation, and investigation of sterol-responsive gene expression. Typical assays encompass cholesterol quantification, SREBP cleavage analysis, RT-qPCR for genes such as HMGCR and LDLR, immunoblotting for FDFT1 and pathway components, proliferation and apoptosis assays, lipidomics, and flow cytometry for lipid raft markers. The cells can also be used in co-culture experiments to assess the impact of altered lipid metabolism on immune cell interactions. For additional technical details or purchasing information, please contact Ascent Research.
