The CLCN6 Knockout Raji Polyclonal Cells are a heterogeneous pool of Raji B lymphocytes with CRISPR/Cas9-mediated disruption of CLCN6, which encodes an endolysosomal voltage-gated chloride channel. This polyclonal knockout population provides a loss-of-function model to study chloride channel function in lysosomal acidification and related processes, preserving genetic diversity absent in monoclonal lines.
The Raji cell line, derived from a Burkitt’s lymphoma patient, is EBV-positive and retains B-cell features including immunoglobulin production and MHC class II antigen presentation. Widely employed in immunology and oncology, Raji cells serve as a robust model for B-cell malignancies and immune signaling, offering a relevant context to examine lysosomal protein functions within antigen-presenting cells.
CLCN6 encodes a voltage-gated chloride channel mediating Cl?/H? exchange in endolysosomal membranes to control luminal pH, a function essential for lysosomal hydrolase activity and autophagy. It is regulated by luminal pH, membrane potential, and chloride levels, and influences downstream targets including TFEB nuclear translocation, autophagosome-lysosome fusion, and mTORC1 activation. The channel interacts with LAMP proteins and the V-ATPase, and its disruption is expected to perturb lysosomal acidification and autophagy flux, impacting pathways involving LC3, LAMP1, and lysosomal hydrolases.
In Raji B cells, lysosomal function is critical for antigen processing and MHC-II-restricted presentation. CLCN6 loss may impair lysosomal proteolysis, altering peptide loading and potentially affecting T-cell responses. Furthermore, autophagy defects could influence cell survival and lymphoma progression. This knockout model thus enables dissection of chloride channel roles in antigen presentation, B-cell lymphoma biology, and lysosomal dysfunction relevant to storage disorders and neurodegeneration.
Applications include lysosomal pH measurement with LysoSensor, autophagy assessment via LC3/p62 western blotting or LC3/LAMP1 immunofluorescence, and antigen presentation assays using MHC-II flow cytometry. The model supports drug screening for lysosomal modulators and exploration of mTORC1/TFEB signaling. RT-qPCR of autophagy genes extends the toolkit. For technical inquiries, contact Ascent Research.
