The NPC2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the Raji B-lymphocyte cell line. This product features targeted disruption of the NPC2 gene, which encodes the lysosomal cholesterol-binding protein NPC2. The polyclonal pool format avoids clonal selection, preserving genetic heterogeneity and providing a robust tool for studying NPC2 loss-of-function. Engineered through CRISPR/Cas9-mediated gene disruption, these cells enable precise interrogation of intracellular cholesterol transport pathways.
Derived from a Burkitt lymphoma patient, the Raji cell line is a widely used human B lymphoblastoid model that retains key characteristics of B lymphocytes, including antigen presentation and immunoglobulin production. As an Epstein-Barr virus (EBV)-immortalized line, Raji cells grow in suspension and are amenable to high-throughput genetic manipulation. Their well-documented cholesterol metabolism pathways make them particularly suitable for investigating lysosomal lipid trafficking and NPC2-dependent processes in a hematopoietic setting.
NPC2 functions as a soluble cholesterol transfer protein within the lysosomal lumen, where it binds cholesterol and delivers it to the NPC1 membrane protein for export to other organelles. This coordinated process, involving interactions with cholesterol and lysosomal integral membrane proteins, maintains cellular cholesterol homeostasis. Disruption of NPC2 abolishes cholesterol transfer to NPC1, leading to lysosomal cholesterol accumulation and downstream metabolic dysregulation. The NPC2/NPC1 axis is a central node in the lysosomal cholesterol egress pathway, and its dysfunction causes Niemann-Pick disease type C2.
Given that B lymphocytes require cholesterol for lipid raft formation, receptor signaling, and antigen presentation, NPC2 knockout in Raji cells is expected to induce lysosomal lipid storage and perturb cholesterol distribution. This model facilitates the study of NPC2 deficiency in immune cells, where altered cholesterol trafficking may impact MHC class II-mediated antigen presentation and B-cell receptor signaling. By providing a tractable system to examine lysosomal dysfunction in a lymphoid context, these cells complement traditional neuronal and hepatic NPC2 models.
Typical applications include Niemann-Pick disease C2 research, lysosomal cholesterol trafficking studies, and drug screening for small molecules that restore cholesterol efflux. The cells are compatible with a range of assays: filipin staining detects unesterified cholesterol accumulation; Amplex Red quantifies total cholesterol; Western blotting and RT-qPCR monitor NPC2 protein and mRNA knockdown; immunofluorescence assesses LAMP1 localization; and cholesterol efflux assays measure export capacity. This polyclonal knockout model offers a versatile human cell platform for mechanistic and therapeutic studies in lysosomal lipid disorders. For additional product information, please contact Ascent Research.
