The NPC1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from human Raji B lymphoblastoid cells. The product is supplied as a mixed population with targeted disruption of the NPC1 gene, preserving genetic heterogeneity without single-cell cloning. CRISPR/Cas9-mediated gene disruption enables efficient ablation of NPC1 protein, creating a versatile loss-of-function model for cholesterol trafficking research.
Raji cells are derived from a Burkitt lymphoma patient and are Epstein-Barr virus (EBV)-positive, offering a robust model for B lymphocyte biology. Widely used to investigate antibody production, antigen presentation, and immune signaling, their rapid growth and established protocols make them suitable for high-throughput genetic and pharmacological studies.
NPC1 encodes a late endosomal/lysosomal cholesterol transporter that works in concert with the soluble cholesterol-binding protein NPC2 and the small GTPase Rab7 to mediate the egress of unesterified cholesterol. Upstream regulators include LDL-derived cholesterol and oxysterols that activate liver X receptor (LXR) pathways. NPC1 loss disrupts this network, suppressing SREBP activation, reducing LDL receptor expression, and inhibiting mTORC1 signaling, while impairing autophagy. This leads to toxic cholesterol accumulation within lysosomes, lysosomal dysfunction, and altered lipid homeostasis, mirroring key pathological features of Niemann-Pick disease type C.
In Raji B lymphoblastoid cells, NPC1 knockout enables dissection of cholesterol trafficking in an immune context. Membrane cholesterol distribution is critical for B cell receptor signaling and antigen presentation; thus, NPC1 loss may perturb immune synapse formation and downstream effector functions. The EBV-transformed background adds a layer of viral manipulation of host lipid metabolism, making this model valuable for studying virus?Chost interactions. Furthermore, the polyclonal nature captures genetic variation, improving relevance for drug response profiling and modeling heterogeneous tumor populations.
This knockout model is ideally suited for research on Niemann-Pick disease type C, lysosomal storage disorders, and therapeutic discovery. Researchers can employ Filipin staining to visualize unesterified cholesterol accumulation, Western blotting to confirm NPC1 ablation, and lysosomal staining to assess organelle changes. Autophagy flux analysis and mTORC1 activity assays provide functional readouts, while flow cytometry-based LDL uptake quantifies trafficking deficits. The model also supports high-throughput screening of small molecules aimed at restoring cholesterol homeostasis. For further technical information, contact Ascent Research.
