The CBARP Knockout Raji Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout population derived from the human Raji B lymphocyte cell line, harboring a targeted disruption of the CBARP gene. This loss?of?function tool permits investigation of CBARP?mediated regulation of voltage?gated calcium channel trafficking and activity within a model of malignant B cells.
The Raji line is an Epstein?CBarr virus?positive Burkitt lymphoma?derived B lymphoblast that carries a t(8;14) MYC/immunoglobulin heavy?chain translocation. It exhibits germinal center B?cell characteristics, expresses functional B?cell receptors, and is capable of antibody production, making it a well?established system for immunological and hematological malignancy research.
CBARP (CACNB?associated regulatory protein) serves as an adaptor that binds to voltage?gated calcium channel ?? subunits (CACNB1?C4) and couples them to the A?kinase anchoring protein AKAP7, thereby localizing PKA to the channel complex. It is regulated by cAMP?dependent PKA, Ca2+/calmodulin?dependent protein kinase II (CaMKII), and ???adrenergic agonists. CBARP modulates the trafficking and surface expression of L?type (CaV1.2, encoded by CACNA1C) and N?type (CaV2.2) channels, thus controlling calcium influx that activates downstream effectors such as calcineurin and CaMKII. In the canonical signaling cascade, ADRB2 stimulation activates GNAS/adenylyl cyclase, generating cAMP that releases PKA catalytic subunits (PRKACA) to phosphorylate CBARP?associated targets, integrating GPCR?cAMP?PKA and calcium signaling.
In Raji B lymphocytes, CBARP disruption is predicted to uncouple PKA signaling from voltage?gated calcium entry, altering B?cell receptor?mediated calcium oscillations and downstream activation events. Dysregulated calcium signaling has been implicated in B?cell lymphoma survival and proliferation; therefore, this knockout model enables dissection of how CBARP?dependent channel modulation impacts malignant B?cell physiology and potentially identifies vulnerabilities in calcium?dependent growth pathways.
Researchers can employ calcium imaging with Fluo?4 to monitor real?time cytosolic Ca2+ dynamics and patch?clamp electrophysiology to assess channel currents. Biochemical characterization via western blotting for CACNB3 and Cav subunits, combined with co?immunoprecipitation, permits analysis of channel?complex composition. Functional studies may include GPCR agonist (isoproterenol) stimulation with phospho?PKA substrate analysis, flow cytometric detection of activation markers (CD69, CD86), and apoptosis or viability assays (Annexin V, MTT). The polyclonal population is also suited for immunological synapse calcium dynamics and drug target validation. For further technical details, please contact Ascent Research.
