The P2RX4 Knockout Raji Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B lymphocyte line, carrying a targeted disruption of the P2RX4 gene. This loss-of-function model provides a robust experimental system for dissecting ATP-gated purinergic signaling in B cell biology without the biases of single-cell cloning. The polyclonal pool retains the heterogeneity of the starting Raji line while enabling unambiguous interrogation of P2RX4-dependent processes. The product is supplied as a ready-to-use knockout resource for advanced biomedical research.
The Raji cell line is an Epstein-Barr virus (EBV)-positive Burkitt lymphoma-derived B lymphocyte line widely employed as a model for B cell biology and lymphomagenesis. Raji cells lack surface immunoglobulin but express characteristic B cell markers, maintaining an immortalized state that supports studies of EBV latency and oncogenic transformation. These cells have been extensively characterized for receptor expression and signaling competence, making them a highly relevant host for examining purinergic receptor functions in malignant B cells.
P2RX4 encodes a ligand-gated cation channel that opens upon binding extracellular ATP, triggering calcium influx and downstream signaling cascades. The channel is activated by ATP and ADP, and its activity can be modulated by kinases including PKC, PKA, and SRC family kinases, as well as by inflammatory cytokines such as TNF-?? and IL-1??. Calcium entry through P2RX4 stimulates calcium/calmodulin-dependent kinases (CaMKII), leading to activation of transcription factors including CREB and NFAT. P2RX4 also interacts with P2RY receptors, Pannexin-1, TLR4, SRC kinases, and caveolin-1, forming a signaling nexus that integrates purinergic and inflammatory pathways. Notably, P2RX4-mediated calcium influx promotes assembly of the NLRP3 inflammasome, resulting in ASC-dependent caspase-1 activation and proteolytic maturation and release of IL-1??, linking ATP sensing to innate immune responses and pain signaling.
In Raji cells, which lack surface immunoglobulin yet retain B cell receptor-related signaling adaptations, P2RX4 knockout provides a unique tool to dissect ATP-driven calcium-dependent pathways in Burkitt lymphoma. Given the role of purinergic signaling in B cell activation and the contribution of NLRP3 inflammasome to lymphoma microenvironment cross-talk, disruption of P2RX4 may uncover mechanisms of lymphomagenesis and EBV latency regulation. Furthermore, because Raji cells are of malignant origin, this model permits investigation of how extracellular nucleotides influence tumor cell viability, immune evasion, and pro-inflammatory cytokine secretion in a B cell malignancy context.
Researchers can employ this knockout model to investigate ATP-mediated signaling in B lymphocytes, assess NLRP3 inflammasome function, evaluate effects on EBV latency, and screen for P2RX4 inhibitors in cancer. Representative assays include Western blotting and RT-qPCR for P2RX4, flow cytometry for surface P2RX4, ATP-induced calcium flux measurement, ELISA for IL-1?? release, Western blotting for NLRP3 and caspase-1, co-immunoprecipitation to probe P2RX4 interactions, phospho-CREB analysis, and cell viability assays. These polyclonal knockout cells thus serve as a versatile platform for studying purinergic signaling in lymphomagenesis and neuroinflammation-associated pathologies. For technical inquiries or additional support, please contact Ascent Research.
