The LONP2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Raji B lymphoblastoid cell line, with targeted disruption of the LONP2 gene. This loss-of-function model is supplied as a heterogeneous pool of edited cells, enabling robust functional studies without clonal selection bias. LONP2 encodes the peroxisomal Lon protease, an ATP-dependent enzyme critical for the selective degradation of oxidized and misfolded proteins within peroxisomes, and its disruption provides a platform to investigate peroxisomal proteostasis and metabolism.
Raji cells are an Epstein-Barr virus (EBV)-positive B lymphoblastoid line derived from a Burkitt lymphoma, exhibiting characteristics of mature B lymphocytes. They serve as a well-established model for B-cell biology, including immunoglobulin synthesis, antigen presentation, and signal transduction, and are extensively employed in immunology and hematological malignancy research. The presence of functional peroxisomes in Raji cells makes them suitable for studying peroxisomal metabolism in the context of lymphocyte physiology, and the LONP2 knockout provides a tool to dissect the role of peroxisomal quality control in these processes.
LONP2 functions as the primary ATP-dependent protease within the peroxisomal matrix, mediating the selective clearance of oxidized and misfolded proteins to maintain peroxisomal proteostasis and optimal fatty acid ??-oxidation. It interacts with PEX5 and PEX14 during substrate import and collaborates with Hsp70 to recognize damaged polypeptides. Key substrates include ACOX1 and D-bifunctional protein; their accumulation upon LONP2 loss impairs very-long-chain fatty acid breakdown. LONP2 is transcriptionally regulated by PPAR?? and PGC-1?? and responsive to reactive oxygen species, positioning it at the intersection of peroxisomal quality control and metabolic signaling.
In Raji B lymphocytes, LONP2 knockout causes accumulation of proteotoxic peroxisomal substrates, impairing fatty acid catabolism and elevating ROS. This metabolic stress perturbs B-cell functions such as survival, proliferation, and antibody production. Given the high metabolic rate of Burkitt lymphoma cells, this model is ideal for studying how peroxisomal dysfunction influences oncogenic pathways. It also mimics features of peroxisomal biogenesis disorders, offering a human B-cell system for disease modeling and therapy assessment.
Researchers can use this knockout model to investigate LONP2-dependent protein quality control by immunoblotting and immunofluorescence for peroxisomal markers catalase and PMP70. Functional readouts??including fatty acid ??-oxidation assays and DCFDA-based ROS detection??reveal metabolic consequences. Apoptosis profiling with Annexin V staining and metabolic flux analysis link peroxisomal defects to cellular bioenergetics. Applications extend to lymphoma drug sensitivity screening and modeling peroxisomal biogenesis disorders. For technical support, contact Ascent Research.
