The PANX1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from Raji B lymphoblastoid cells, designed to disrupt the PANX1 gene. This heterogeneous pool of edited alleles provides a robust loss-of-function model without clonal selection, enabling studies of PANX1-dependent processes in a lymphoid background with reduced clonal bias.
Raji cells originate from an EBV-positive Burkitt lymphoma and serve as a well-established model for B lymphocyte biology, EBV latency, and lymphomagenesis. Their lymphoblastoid phenotype, high proliferative capacity, and expression of key purinergic receptors make them highly suitable for gene disruption experiments aimed at dissecting signaling pathways relevant to hematological malignancies and immune cell communication.
PANX1 encodes a large-pore channel that mediates ATP release, functioning as a hemichannel and, in some settings, forming gap junctions. Its activity is triggered by extracellular ATP via the P2X7 receptor, mechanical stress, and by caspase-3/7 cleavage during apoptosis. Downstream, PANX1-mediated ATP egress activates purinergic P2Y/P2X receptors, driving NLRP3 inflammasome assembly with ASC, leading to caspase-1 activation and IL-1?? secretion. PANX1 interacts with actin and assembles into homomeric complexes. In apoptotic cells, caspase cleavage of PANX1 sustains ATP release, generating a ??find-me?? signal for phagocyte recruitment.
In Raji B cells, PANX1 knockout disrupts ATP release and purinergic signaling, allowing precise dissection of inflammasome activation and apoptotic clearance mechanisms. The cells express endogenous purinergic receptors and inflammasome components, providing a relevant context to study how PANX1 deficiency influences the NLRP3/IL-1?? axis, hemichannel-mediated communication, and tumor microenvironment interactions. This model also offers a non-neuronal platform for studying PANX1-associated neuropathic pain and migraine pathways.
Applications include quantitative ATP release luciferase assays, dye uptake measurements, IL-1?? ELISA, caspase-1 activity assays, western blot, RT-qPCR, and immunofluorescence. The polyclonal knockout cells support drug screening for PANX1 modulators, apoptosis signaling studies, cancer cell communication research, and investigation of ischemia-reperfusion injury. For technical assistance and custom experimental design, contact Ascent Research.
