The PANK2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from Raji B lymphocytes, with targeted disruption of the PANK2 gene. This loss-of-function model eliminates mitochondrial pantothenate kinase 2, enabling investigation of CoA biosynthesis deficiency without monoclonal selection.
Raji cells, an EBV-positive Burkitt lymphoma line, serve as a widely used model for B-cell functions and oncogenesis. The integration of PANK2 knockout into this immortalized background allows interrogation of metabolic pathways within malignant B lymphocytes, expanding studies of PKAN-related mitochondrial dysfunction to a hematological context.
PANK2 catalyzes the rate-limiting phosphorylation of pantothenate to 4′-phosphopantothenate in mitochondrial CoA synthesis. It is regulated by insulin/IGF-1, mTORC1, PPARA, and feedback inhibition by CoA/acetyl-CoA, and interacts with COASY, PPCS, cardiolipin, and NCLX, forming homodimers. Downstream targets include CoA, acetyl-CoA, succinyl-CoA, propionyl-CoA, and mitochondrial respiratory chain complexes, linking to TCA cycle and histone acetylation. PANK2 disruption impairs CoA production, fatty acid oxidation, and oxidative phosphorylation, promoting mitochondrial dysfunction, oxidative stress, and iron dyshomeostasis reminiscent of PKAN.
In Raji B cells, PANK2 knockout affects energy metabolism and CoA-dependent processes, offering a tool to study B-cell metabolism, lymphoma biology, and neurodegeneration mechanisms. The model exhibits iron accumulation and lipid peroxidation, measurable by Perls’ staining and C11-BODIPY, mimicking PKAN pathology and enabling translational research in a non-neuronal system, while also elucidating metabolic reprogramming in lymphoma.
Applications include elucidating CoA deficiency in B cells, modeling PKAN, screening small molecules to restore CoA, studying lipid metabolism and oxidative stress in lymphoma, and investigating mitophagy and ferroptosis. Assays encompass Western blotting, RT-qPCR, LC-MS for CoA/acetyl-CoA, Seahorse respiration, Perls’ staining, C11-BODIPY, TMRE flow cytometry, apoptosis detection, and viability under pantothenate deprivation. Contact Ascent Research for details.
