The CLCN2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Raji B lymphocyte line, featuring disruption of the CLCN2 gene that encodes the voltage-gated chloride channel ClC-2. This loss-of-function model provides a tool to investigate chloride transport, cell volume regulation, and related signaling in a human Burkitt??s lymphoma context. As a polyclonal pool, it reflects genetic heterogeneity, avoiding clonal selection artifacts and enabling robust functional studies.
The Raji host cell line is an EBV-positive Burkitt??s lymphoma-derived B lymphocyte line widely used to model B cell biology and lymphomagenesis. These suspension-adapted cells maintain key pathways of humoral immune responses and are amenable to electrophysiological recordings and high-throughput applications. The EBV-driven proliferation sustains a physiological environment for studying ion channel function.
CLCN2 encodes ClC-2, a voltage-gated chloride channel activated by hyperpolarization, cell swelling, and acidic pH. SGK1 kinase phosphorylates and enhances ClC-2 activity, while NEDD4-2 ubiquitinates it for internalization. The channel interacts with HEPACAM and the actin cytoskeleton and is modulated by AMPK and PI3K/AKT signaling. Downstream, ClC-2 mediates chloride efflux for regulatory volume decrease and apoptotic volume decrease, linking cell volume homeostasis to apoptosis.
In Raji B lymphocytes, ClC-2 contributes to volume regulation during proliferation and apoptosis. Cell swelling triggers SGK1-mediated ClC-2 activation, promoting volume recovery. Disruption of CLCN2 may impair volume homeostasis and alter apoptotic sensitivity, offering insights into chloride channel roles in B cell survival and immune function. This model also aids study of ClC-2-related channelopathies such as leukoencephalopathy and epilepsy.
These knockout cells enable patch clamp analysis of chloride currents, cell volume measurements by Coulter counter, and flow cytometric apoptosis assays with Annexin V/PI. Western blot, RT-qPCR, and immunofluorescence confirm CLCN2 disruption, while chloride-sensitive dyes assess channel function. They are suitable for screening ClC-2 modulators, investigating B cell activation, and performing proliferation assays. For additional information, please contact Ascent Research.
