The CYP51A1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-mediated polyclonal knockout population with targeted disruption of the CYP51A1 gene in human Raji B lymphoblastoid cells. This heterogeneous pool avoids clonal artifacts and captures population-level loss-of-function effects, making it suitable for bulk phenotypic analyses of cholesterol metabolism and signaling.
Raji cells, derived from a Burkitt lymphoma patient, are Epstein-Barr virus (EBV)-positive and grow in suspension. They are widely used to study immune response, antibody production, and antigen presentation, providing a robust model for investigating B-cell biology and lymphomagenesis. Their cholesterol-dependent membrane properties make them particularly relevant for sterol pathway disruption.
CYP51A1 encodes lanosterol 14??-demethylase, which catalyzes the 14??-demethylation of lanosterol in cholesterol biosynthesis. This step is regulated upstream by SREBP2, SCAP, INSIG1, and LXR in response to cholesterol levels, and requires cytochrome P450 reductase (POR) and cytochrome b5. CYP51A1 acts downstream of squalene epoxidase (SQLE) and lanosterol synthase (LSS), and upstream of DHCR7 and DHCR24. Knockout disrupts cholesterol production, causing accumulation of 14??-methyl sterols and impairing membrane integrity, lipid raft signaling, and synthesis of bile acids and steroid hormones.
In Raji cells, CYP51A1 knockout creates a critical model for exploring cholesterol dependency in B-cell proliferation and survival. The loss of functional enzyme unmasks potential vulnerabilities to statins or azole antifungals and enables studies of sterol-related disorders such as congenital cataracts and microcephaly. This system also allows interrogation of SREBP2/LXR feedback loops and adaptive metabolic responses, linking sterol metabolism to immune cell function.
Applications include Western blotting for CYP51A1, RT-qPCR for sterol gene expression (e.g., HMGCR, DHCR7), and Amplex Red cholesterol quantification. Cell proliferation, apoptosis, and drug sensitivity assays can assess statin and azole effects. These cells are valuable for drug target validation and mechanistic studies in cholesterol biology. For further information, contact Ascent Research.
