The LRRC8A Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B lymphoblastoid cell line, designed for functional loss-of-function studies of the LRRC8A gene. This product consists of a heterogeneous mixture of cells carrying targeted disruptions in the LRRC8A locus, providing a robust model for investigating volume-regulated anion channel (VRAC) biology without the influence of wild-type protein. The polyclonal format facilitates rapid experimental deployment while maintaining sufficient knockout penetrance for most cell-population-based assays.
The parental Raji cell line is a well-characterized human B lymphocyte model established from an Epstein-Barr virus (EBV)-positive Burkitt lymphoma. These cells express classical B cell markers such as CD19, CD20, CD22, and surface immunoglobulin M (IgM), and they retain key features of B cell signaling and apoptosis. Raji cells are extensively used in cancer research, immunological studies, and drug development, making them a relevant host for dissecting molecular mechanisms underlying B cell malignancies, immune responses, and volume homeostasis.
LRRC8A encodes a crucial subunit of VRAC, which forms heteromeric channels with other LRRC8 family members (LRRC8B?CE). These channels are activated by cell swelling in response to osmotic stress and are regulated by intracellular ATP, reactive oxygen species, and phosphorylation by Src family kinases and protein kinase C alpha (PKC??). Upon activation, VRAC mediates chloride efflux and organic osmolyte transport, driving regulatory volume decrease (RVD). Additionally, LRRC8A-containing channels facilitate ATP release and membrane potential depolarization, linking volume regulation to purinergic signaling. The protein interacts with integrin beta 1 (ITGB1) and growth factor receptor-bound protein 2 (GRB2), integrating volume sensing with cytoskeletal dynamics and downstream signaling cascades. Disruption of LRRC8A impairs these processes, offering a platform to study VRAC-dependent cellular functions.
In Raji B lymphocytes, LRRC8A-mediated volume regulation is critical for cell survival during osmotic challenges encountered in lymphoid tissues and during immune responses. Loss of LRRC8A permits direct examination of apoptotic volume decrease (AVD) and its connection to apoptosis induction. Furthermore, VRAC activity has been linked to chemoresistance against cisplatin and doxorubicin, positioning this knockout model for drug resistance studies relevant to lymphoma therapy. The association of LRRC8A mutations with agammaglobulinemia type 3 and B-cell deficiencies further highlights the model’s translational relevance for immunodeficiency research.
Researchers can utilize these cells in a variety of assays: whole-cell patch clamp electrophysiology to measure VRAC currents under hypotonic stimulation, flow cytometry or Coulter-based cell volume analyses to monitor RVD kinetics, and Western blotting or RT-qPCR to confirm LRRC8A disruption. The cells are also suitable for Annexin V-based apoptosis assays and drug sensitivity screening with cisplatin or doxorubicin. Additionally, high-throughput screening of VRAC modulators can be performed to identify channel inhibitors or activators. These applications facilitate mechanistic studies of lymphocyte volume control, purinergic signaling, and drug resistance mechanisms. For further technical information, please contact Ascent Research.
