The COX7A2L Knockout Raji Polyclonal Cells constitute a CRISPR/Cas9-mediated loss-of-function model in which the COX7A2L gene has been disrupted in a polyclonal population of Raji B lymphocytes. This polyclonal knockout cell pool, generated through CRISPR/Cas9 gene editing, provides a heterogeneous collection of edited cells for studying COX7A2L-dependent functions without clonal selection. The product enables investigation of cytochrome c oxidase (Complex IV) biology in a human B-cell background, offering a versatile tool for mitochondrial respiratory chain research.
The Raji host cell line is derived from a Burkitt lymphoma patient and is Epstein-Barr virus (EBV)-positive, exhibiting a lymphoblastoid phenotype. As a B-lymphocyte cell line, Raji cells are widely employed to study antibody production, antigen presentation, and adaptive immune responses. Their transformed nature and robust growth characteristics facilitate biochemical and functional analyses of mitochondrial respiration, making them a suitable host for interrogating the metabolic dependencies of lymphoma-derived cells.
COX7A2L encodes a subunit of cytochrome c oxidase (Complex IV) that is essential for electron transfer from cytochrome c to oxygen, coupled with proton translocation across the inner mitochondrial membrane. The protein interacts with core Complex IV subunits COX1, COX2, and COX3, as well as with assembly factors such as SCO1, SCO2, COX10, and COX15. COX7A2L expression is regulated by transcriptional activators PPARGC1A (PGC-1??), NRF1, TFAM, and ESRRA, while being repressed by HIF1A under hypoxia. Loss of COX7A2L disrupts supercomplex assembly, impairing electron flow through NADH dehydrogenase (Complex I), succinate dehydrogenase (Complex II), cytochrome bc1 (Complex III), and ultimately cytochrome c oxidase (Complex IV), leading to diminished ATP synthesis by ATP synthase (Complex V), elevated reactive oxygen species (ROS) production, and altered apoptosis signaling.
In the Raji B-cell context, COX7A2L knockout is expected to compromise oxidative phosphorylation, forcing a metabolic shift that mimics cancer metabolic reprogramming frequently observed in lymphomas. This model system enables dissection of mitochondrial dysfunction in malignant B cells, shedding light on how Complex IV defects affect cell viability, proliferation, and sensitivity to chemotherapeutic agents. By altering mitochondrial membrane potential and ROS levels, the knockout may modulate B-cell receptor signaling and apoptotic thresholds, providing insights into lymphoma pathogenesis.
Researchers can use these polyclonal knockout cells for cancer metabolism studies, mitochondrial dysfunction analysis in lymphoma, and oxidative phosphorylation profiling in B lymphocytes. Functional assays include oxygen consumption rate (OCR) measurement, Complex IV activity assay, ATP quantification, mitochondrial membrane potential analysis (JC-1), ROS detection (DCFDA), and apoptosis assays. Western blotting and RT-qPCR validate COX7A2L disruption, while metabolomic profiling reveals metabolic shifts. For further information, contact Ascent Research.
