The MSMO1 Knockout Raji Polyclonal Cells are a polyclonal population of Raji B lymphocytes engineered via CRISPR/Cas9 to disrupt functional MSMO1 expression. This viable polyclonal knockout pool enables direct loss-of-function analysis without single-cell cloning. CRISPR editing introduces heterogeneous MSMO1 locus disruptions, collectively eliminating C4-methyl sterol oxidase activity. This model suits investigations of cholesterol metabolism in B-cell contexts where lipid pathways are critical for proliferation and survival.
The parental Raji line is a human Burkitt lymphoma B lymphoblast that harbors EBV and serves as a model for B-cell malignancies. These cells display germinal center B-cell features and are used in immunology, hematologic oncology, and drug discovery. The EBV-positive background elevates de novo cholesterol biosynthesis to maintain viral latency and tumor growth. Thus, Raji cells offer a clinically relevant system to study cholesterol metabolism?Clymphoma interactions.
MSMO1 encodes methylsterol monooxygenase 1, which catalyzes C4-methyl oxidation in post-lanosterol cholesterol synthesis. It acts downstream of LSS and CYP51A1, and upstream of NSDHL, SC5D, and DHCR7. The SREBF2?CSCAP?CInsig1/2 axis transcriptionally regulates MSMO1 in response to low sterol levels, alongside HMGCR and SQLE. MSMO1 knockout causes methylsterol intermediate accumulation and cholesterol depletion, impairing synthesis of 7-dehydrocholesterol, desmosterol, cholesteryl esters, and oxysterols.
In Raji lymphoma, MSMO1 loss creates metabolic vulnerability because these cells depend on cholesterol for membrane biogenesis, lipid rafts, and signaling. This polyclonal knockout model enables study of cholesterol auxotrophy in B-cell cancer, compensatory adaptations, and synthetic lethality approaches. It also permits investigation of how intrinsic cholesterol production affects B-cell receptor signaling and EBV latency, revealing tumor-specific lipid dependencies.
These cells support diverse applications including cholesterol metabolic flux analysis, lipidomic profiling of methylsterol intermediates, and drug sensitivity screening against statins or cholesterol auxotrophy-inducing agents. Compatible assays encompass filipin staining for unesterified cholesterol, immunoblotting, RT-qPCR, flow cytometry for lipid raft markers, and proliferation assays under lipid-depleted conditions. The polyclonal nature closely mimics tumor genetic heterogeneity, providing a physiologically relevant system for functional genomics and therapeutic evaluation. For further technical information, please contact Ascent Research.
