The DHCR24 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the DHCR24 gene has been disrupted to eliminate expression of 3??-hydroxysterol ??24-reductase, the enzyme that catalyzes the terminal step of the Bloch pathway, converting desmosterol to cholesterol. This polyclonal knockout pool, derived through CRISPR/Cas9-mediated gene targeting, provides a robust loss-of-function model without the constraints of single-cell cloning, enabling reproducible functional genomics studies across a heterogeneous genetic background.
Raji cells, the host line for this knockout, are an EBV-positive B lymphoblast line originally isolated from a patient with Burkitt lymphoma. These suspension-adapted cells exhibit mature B-cell characteristics and are widely employed in immunological research, particularly for investigating B-cell receptor signaling, apoptosis, and viral oncogenesis. Their rapid proliferation and well-characterized signaling networks make them an ideal platform for dissecting lipid-dependent processes in hematopoietic malignancies.
DHCR24 functions at a critical node in cholesterol biosynthesis, acting downstream of HMGCR, SQLE, LSS, CYP51A1, and MSMO1. Its expression is tightly regulated by SREBP2 in response to sterol levels, with additional modulation by INSIG, LXR, and p53. The reductase physically interacts with caveolin-1, APP, Mdm2, and DHCR7, and its product cholesterol is essential for lipid raft organization, steroid hormone synthesis, and protein prenylation. By controlling membrane microdomain architecture, DHCR24 influences signal transduction, oxidative stress responses, and cell survival, linking lipid metabolism to p53 and Hedgehog pathway activity.
In Raji B lymphocytes, ablation of DHCR24 disrupts cholesterol homeostasis, leading to altered membrane fluidity and lipid raft composition. This perturbation can impair B-cell receptor clustering and downstream kinase cascades, while also affecting EBV-mediated oncogenic programs, as viral entry and latent protein localization depend on cholesterol-rich domains. Such metabolic reprogramming sensitizes cells to oxidative stress and pro-apoptotic stimuli, creating a powerful model to explore the vulnerabilities of lymphoma cells and the interplay between lipid metabolism and immune cell signaling.
This DHCR24 knockout Raji pool supports diverse research applications, including cholesterol metabolic studies via enzymatic quantification and filipin staining, lipid raft analysis through flow cytometry and lipidomics, and signaling pathway dissection using Western blot and RT-qPCR. It enables functional assays such as apoptosis and cell viability tests under cholesterol depletion, co-immunoprecipitation for protein interaction studies, and SREBP2 transcriptional reporter assays. These tools facilitate investigations into desmosterolosis, Alzheimer??s disease, hepatocellular carcinoma, prostate cancer, melanoma, and metabolic syndrome. For further technical details or ordering information, please contact Ascent Research.
