PHKA1 Knockout Raji Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of Raji B lymphocytes with targeted disruption of the PHKA1 gene. This product provides a genetically heterogeneous pool bearing diverse loss-of-function alleles, generated by transient delivery of Cas9 and a specific guide RNA, followed by selection to enrich for edited cells. The polyclonal format minimizes clonal selection bias and preserves phenotypic variability, making it suitable for studies requiring bulk cell behavior. The knockout leads to ablation of the phosphorylase kinase alpha regulatory subunit, impairing functional enzyme complex assembly.
The Raji parental line is a human Burkitt??s lymphoma B-cell line, isolated from a patient with non-Hodgkin lymphoma and characterized by EBV positivity. Raji cells serve as a widely used model for B-cell malignancies, antigen presentation, and EBV biology, exhibiting constitutive NF-??B activity and metabolic adaptations typical of lymphoma. Their robust growth and well-characterized signaling networks facilitate investigation of gene function in a lymphoma context.
PHKA1 encodes the alpha subunit of phosphorylase kinase, which activates glycogen phosphorylase (PYGL and PYGB) to mobilize glucose from glycogen. The enzyme complex is regulated by calcium through calmodulin (CALM1) and by cAMP-dependent protein kinase A (PKA) downstream of ??-adrenergic receptors. The alpha subunit provides a scaffold for beta (PHKB) and gamma (PHKG1) subunits, integrating hormonal and metabolic signals. Therefore, PHKA1 functions upstream of PYGL/PYGB and is essential for glycogenolysis, bridging calcium and PKA signaling to glucose-1-phosphate production. Knocking out PHKA1 disrupts complex formation, preventing phosphorylation of glycogen phosphorylase and halting glycogen breakdown.
In the Raji B-cell lymphoma background, PHKA1 knockout offers a model to study metabolic dependencies of malignant B cells. Lymphoma cells often rewire metabolism to support proliferation; glycogen can serve as a carbon reserve or contribute to nucleotide sugar synthesis. Loss of glycogenolysis may force reliance on alternative energy sources and sensitize cells to metabolic stress. Moreover, since phosphorylase kinase is a calcium-responsive enzyme, the knockout allows dissection of calcium signaling cross-talk with glycogen metabolism in apoptosis and immune signaling. This model also permits exploration of glycogen storage disease type IXd, caused by PHKA1 deficiency, in a human B-cell context.
Representative applications include metabolic flux analysis using glucose release assays and glycogen content quantification, complemented by PAS staining. The cells are suitable for Western blotting to verify PHKA1 loss and assess associated subunits, RT-qPCR for transcript monitoring, and phosphorylase kinase activity assays. Calcium flux experiments and apoptosis assays can examine the impact on cell death pathways. For further information, please contact Ascent Research.
