NR1H2 Knockout Raji Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population of Raji B lymphocytes with targeted disruption of the NR1H2 gene, encoding liver X receptor beta (LXR??). This polyclonal knockout model provides a heterogeneous pool of cells carrying CRISPR/Cas9-mediated loss-of-function mutations in NR1H2, enabling functional studies of LXR?? signaling in a B-cell lymphoma context without requiring clonal isolation.
The Raji cell line is an Epstein-Barr virus (EBV)-positive Burkitt lymphoma line with lymphoblastoid morphology, cultured in suspension. Originating from a patient with Burkitt lymphoma, Raji cells serve as a well-characterized model for B-cell biology, lymphomagenesis, and immune function. These immortalized B lymphocytes retain key signaling pathways relevant to cholesterol metabolism and inflammatory responses, making them a suitable host for dissecting the role of LXR?? in lipid homeostasis and B-cell pathophysiology.
NR1H2 (LXR??) encodes a ligand-activated nuclear receptor that heterodimerizes with retinoid X receptor (RXR??/??/??) to regulate gene expression. Upon binding of oxysterol ligands such as 22R-hydroxycholesterol and 24S-hydroxycholesterol, or synthetic agonists like T0901317 and GW3965, the LXR??/RXR complex activates transcription of target genes including ABCA1 and ABCG1, which mediate cholesterol efflux, SREBF1 and FASN involved in fatty acid synthesis, and APOE for lipid transport. NR1H2 also modulates inflammatory responses via transrepression of NF-??B p65 and AP-1, influencing cytokines like TNF and IL10. Coactivators NCOA1 (SRC-1) and PGC-1??, along with corepressors NCOR1 and SMRT, fine-tune LXR?? transcriptional activity.
In Raji B cells, NR1H2 disruption compromises the LXR/RXR transcriptional program, potentially altering cholesterol homeostasis, lipid metabolism, and immune-related gene expression. Loss of LXR?? function can reduce expression of cholesterol transporters ABCA1 and ABCG1, leading to intracellular lipid accumulation and membrane remodeling, which may impact B-cell receptor signaling and proliferation. Furthermore, dysregulation of targets such as MYC and BCL2 may affect cell survival and lymphomagenesis. This polyclonal knockout population provides a physiologically relevant system to study how LXR??-dependent pathways intersect with B-cell malignancies and metabolic reprogramming.
Researchers can employ these knockout cells in a variety of assays to dissect LXR?? function in B lymphocytes. Western blotting and RT-qPCR confirm NR1H2 loss and altered expression of downstream targets like ABCA1, ABCG1, and SREBF1. Cholesterol efflux assays and filipin staining assess lipid transport and intracellular cholesterol accumulation. Proliferation (MTT/BrdU) and apoptosis (Annexin V/7-AAD) assays evaluate the impact of NR1H2 disruption on cell growth and survival, while RNA-seq and ChIP-qPCR profile transcriptomic changes and LXR?? binding sites. Flow cytometry permits analysis of surface markers and lipid raft integrity. Treatment with LXR agonists (T0901317, GW3965) or antagonists enables pharmacological studies. This model is a valuable tool for atherosclerosis, type 2 diabetes, obesity, Alzheimer’s disease, and lymphoma research. For further information, please contact Ascent Research.
