The CLCN7 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of Raji B lymphocytes featuring targeted disruption of the CLCN7 gene. This heterogeneous knockout model avoids clonal artifacts while ensuring efficient gene depletion across the population, enabling robust loss-of-function studies in a B?lymphocyte background. The polyclonal format preserves the genetic diversity of the parental line, making it suitable for functional assays that require population-level readouts.
The parental Raji cell line is a human Burkitt??s lymphoma-derived B lymphoblastoid line that is Epstein-Barr virus (EBV) positive. Raji cells are widely used to study B lymphocyte biology, including antibody production, antigen presentation, and immune responses. Their malignant B cell background makes them a relevant model for B cell malignancies, where lysosomal and autophagy pathways play critical roles in cell survival and drug resistance. CLCN7 knockout in this context offers a unique system to investigate these processes in a clinically relevant setting.
CLCN7 encodes a voltage-gated Cl?C/H+ antiporter that forms a complex with OSTM1 on lysosomal and osteoclast ruffled border membranes. This complex collaborates with the V-ATPase to maintain acidic pH, essential for lysosomal hydrolase activity and autophagy flux. CLCN7 expression is regulated by MITF, TFE3, and TFEB, downstream of mTORC1 and RANKL signaling. Knockout disrupts Cl?C/H+ exchange, impairing lysosomal acidification, reducing cathepsin activity, and blocking autophagy, evidenced by decreased LC3-II turnover. Downstream effects include altered lysosomal pH and degradative maturation.
In Raji B lymphocytes, CLCN7 knockout compromises lysosomal function and autophagy, processes critical for protein turnover and metabolic adaptation. As B cell malignancies often depend on autophagy for survival under stress, CLCN7 disruption may sensitize cells to apoptosis or chemotherapeutics. This polyclonal knockout population enables studies on lysosomal acidification in B cell proliferation, immune signaling, and tumorigenesis without clonal bias. It also provides a model for exploring osteopetrosis-related lysosomal dysfunction in a hematopoietic environment.
Applications include lysosomal dysfunction modeling, autophagy research, and drug screening for B cell malignancies. Key assays are western blotting for CLCN7 and OSTM1, immunofluorescence for LAMP1/2, LysoSensor pH measurement, cathepsin activity, LC3-II turnover, flow cytometry for apoptosis, and RT-qPCR for autophagy genes. These tools facilitate studies of osteopetrosis, lysosomal storage disorders, and neurodegeneration in a hematopoietic context. For more information, please contact Ascent Research.
