The CYB5R1 Knockout Raji Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population of Raji B lymphocytes carrying a targeted disruption of the CYB5R1 gene, providing a loss-of-function model for investigating NADH-dependent cytochrome b5 reductase 1 in a human lymphoblastoid background. This polyclonal knockout pool retains the genetic heterogeneity inherent to a mixed edited population, enabling robust functional studies without clonal selection artifacts, while avoiding the constraints of single-cell-derived lines. The CRISPR/Cas9-mediated gene disruption eliminates full-length CYB5R1 protein expression, generating a null allele that abolishes canonical electron transfer from NADH to cytochrome b5, thereby impairing downstream microsomal redox reactions.
The host Raji cell line is an immortalized male B lymphocyte line derived from a Nigerian patient with Burkitt’s lymphoma, latently infected with Epstein-Barr virus (EBV). Widely employed to model EBV-driven lymphomagenesis and immune cell interactions, Raji cells exhibit a lymphoblastoid phenotype and maintain expression of B-cell surface markers, making them a suitable platform for studying B-cell biology and oncogenic transformation. Their EBV-positive status permits exploration of viral?Chost metabolic crosstalk, while their rapid proliferation facilitates high-throughput functional genomics and pharmacological screening in a lymphoma context.
CYB5R1 encodes an NADH-dependent cytochrome b5 reductase that catalyzes the transfer of electrons from NADH to cytochrome b5 (CYB5A), a key electron donor for microsomal desaturases, elongases, and cytochrome P450 enzymes. As part of the electron transport chain on the cytoplasmic surface of the endoplasmic reticulum, CYB5R1 directly interacts with CYB5A and functionally partners with downstream effectors such as stearoyl-CoA desaturase (SCD1), fatty acid desaturase 2 (FADS2), and CYP51A1. Its activity is transcriptionally regulated by SREBP transcription factors and peroxisome proliferator-activated receptors (PPARs), and it is modulated by oxidative stress response elements. Disruption of CYB5R1 therefore uncouples NADH oxidation from cytochrome b5 reduction, perturbing fatty acid desaturation, cholesterol biosynthesis, and phase I drug metabolism, while altering cellular NADH/NAD+ ratios.
In the Raji lymphoblast model, loss of CYB5R1 is expected to dysregulate lipid remodeling essential for membrane biogenesis and signaling in rapidly dividing B cells. Given the reliance of lymphoma cells on fatty acid and cholesterol synthesis for proliferation, CYB5R1 knockout may compromise the production of unsaturated fatty acids and sterol intermediates, potentially affecting lipid raft integrity, EBV latent gene expression, and B-cell receptor signaling. This model also provides a tool to dissect how redox imbalances driven by CYB5R1 deficiency influence immune cell survival, apoptosis susceptibility, and metabolic adaptation in a cancer cell context.
Key applications include mechanistic dissection of lipid metabolism in B-cell lymphoma, evaluation of cytochrome b5 reductase dependency in EBV-transformed cells, and assessment of chemosensitivity mediated by microsomal P450 enzymes. Experimentally, the model supports Western blotting and RT-qPCR for knockout validation, GC-MS fatty acid profiling, quantitative cholesterol measurement, NADH/NAD+ ratio assays, and flow cytometric analysis of proliferation and apoptosis. These approaches enable systematic investigation of CYB5R1??s role in lymphomagenesis, metabolic reprogramming, and immune cell function. For additional information or technical support, please contact Ascent Research.
