CYB5A Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the human Burkitt lymphoma Raji cell line, in which the CYB5A gene has been disrupted to create a loss-of-function model. This polyclonal knockout product provides a heterogeneous pool of cells carrying targeted mutations, enabling experimental analysis without the bottleneck of clonal isolation. The knockout disrupts the endogenous cytochrome b5 function, allowing researchers to study the immediate and downstream consequences of CYB5A ablation in a B-lymphocyte context.
The Raji cell line, established from an Epstein-Barr virus (EBV)-positive Burkitt lymphoma patient, is a widely used model in immunology and cancer biology. These B lymphocytes express characteristic surface markers including CD19, CD20, CD22, and CD79a, while lacking surface immunoglobulin, reflecting their mature B-cell phenotype. Their EBV-driven transformation and well-defined genomic landscape make Raji cells particularly suitable for investigating the interplay between oncogenic signaling and metabolic pathways in B-cell malignancies.
The CYB5A gene encodes cytochrome b5 type A, a hemoprotein anchored to the endoplasmic reticulum and mitochondrial outer membrane. In microsomal redox reactions, cytochrome b5 accepts electrons from NADH-cytochrome b5 reductase (CYB5R3) and delivers them to critical acceptors such as the fatty acid desaturases SCD, FADS1, and FADS2, and cytochrome P450 enzymes CYP17A1, CYP21A2, and CYP19A1. This electron transfer is essential for fatty acid desaturation, cholesterol biosynthesis, and steroid hormone hydroxylation. CYB5A expression is transcriptionally regulated by SREBF1, HNF4A, and NFE2L2, linking its activity to lipid homeostasis and oxidative stress responses. Disruption of CYB5A in these polyclonal cells severs the electron supply, causing potential deficiencies in lipid desaturation and steroidogenesis.
Within the Raji B-lymphocyte model, ablation of CYB5A provides a valuable system to explore the role of microsomal redox regulation in immune cell function and malignant transformation. B cells depend on membrane lipid composition for receptor signaling, proliferation, and survival; thus, impaired fatty acid desaturation may alter raft formation and downstream signaling dynamics. Additionally, cytochrome b5 participates in the metabolism of various chemotherapeutic agents through P450 enzymes, making this knockout population a relevant tool for studying drug sensitivity and metabolism in lymphomas. The model also aids research into methemoglobin reduction and congenital disorders linked to cytochrome b5 deficiency.
Investigators can apply CYB5A Knockout Raji Polyclonal Cells in a variety of downstream assays. Lipidomic analysis can identify shifts in fatty acid saturation profiles, while cytochrome b5 reductase activity assays and drug metabolism experiments with P450 substrates quantify functional impairment. Flow cytometry enables detection of apoptosis and B-cell marker expression, and RT-qPCR or Western blotting can validate pathway alterations at the molecular level. These cells are particularly suited for studying steroidogenic capacity in immune cells and redox biology in cancer. For additional information or ordering, please contact Ascent Research.
