The DNAJC6 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Burkitt’s lymphoma B lymphocyte line Raji, in which the DNAJC6 gene has been disrupted to abolish expression of functional auxilin. This polyclonal population retains diverse editing events across cells, providing a heterogeneous loss-of-function model that avoids clonal artifacts and reflects the complexity of gene disruption at the population level.
Raji cells are an Epstein-Barr virus (EBV)-positive, suspension-growing B lymphocyte line established from a Burkitt’s lymphoma. They carry the characteristic t(8;14) translocation that juxtaposes the c-MYC oncogene with the immunoglobulin heavy-chain locus, resulting in constitutive c-MYC overexpression. Widely employed as a model for B cell biology and lymphomagenesis, Raji cells express surface B cell receptors, transferrin receptors, and various signaling receptors, making them a robust platform for investigating endocytic trafficking and membrane dynamics.
The DNAJC6 gene encodes auxilin, a J-domain?Ccontaining co-chaperone that recruits Hsc70 (HSPA8) to clathrin-coated vesicles. Auxilin stimulates Hsc70??s ATPase activity, driving clathrin lattice disassembly after endocytosis. In clathrin-mediated endocytosis, auxilin functions downstream of receptor activation by ligands such as transferrin and EGF, which engage the AP2 adaptor complex and clathrin triskelion at the plasma membrane. Auxilin interacts directly with clathrin heavy chain and Hsc70, and its activity is essential for uncoating of endocytic vesicles carrying transferrin receptor and epidermal growth factor receptor. Dynamin mediates vesicle scission upstream of auxilin, completing the cycle of receptor internalization and clathrin recycling.
Knockout of DNAJC6 in Raji cells is predicted to impair clathrin-mediated endocytosis by blocking vesicle uncoating, leading to accumulation of coated pits, altered surface receptor levels, and reduced internalization of ligands such as transferrin and EGF. This defect likely impacts B cell receptor trafficking and downstream signaling pathways that depend on receptor endocytosis for attenuation or sustained activation. As mutations in DNAJC6 are linked to autosomal recessive juvenile Parkinson disease (PARK19), this knockout model provides a unique intersection between neurodegenerative disease mechanisms and B lymphocyte cell biology, enabling studies of auxilin??s role in endocytic homeostasis.
Researchers can employ the DNAJC6 Knockout Raji Polyclonal Cells as a versatile tool to dissect clathrin-mediated endocytosis and receptor trafficking. Representative applications include transferrin uptake assays to quantify endocytic efficiency, EGF internalization kinetics, immunofluorescence microscopy to visualize clathrin coat accumulation, flow-cytometric measurement of surface transferrin receptor and EGFR levels, and co-immunoprecipitation of Hsc70?Cclathrin complexes. The model also supports investigations into drug delivery mechanisms via endocytosis and can serve as a cellular system to study molecular events underlying Parkinson??s disease?Crelated neurodegeneration. For additional information or to discuss custom gene-editing projects, please contact Ascent Research.
