The CMC2 Knockout Raji Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Raji B lymphocyte line, engineered to disrupt the CMC2 gene. This loss-of-function model is specifically designed for advanced studies of mitochondrial cytochrome c oxidase (COX) assembly and copper homeostasis within a physiologically relevant lymphoblastoid background. The polyclonal format provides a heterogeneous knockout cell pool, enabling robust functional analyses without clonal selection biases, and is suitable for investigating bulk cellular responses to CMC2 deficiency.
Raji cells are an immortalized B lymphocyte line originally isolated from a Burkitt lymphoma patient. These cells are Epstein-Barr virus (EBV) positive and exhibit characteristic B-cell surface markers, including CD19 and CD20, making them a widely accepted model for B-cell malignancies. Their lymphoblastoid nature facilitates studies of B-cell receptor signaling, lymphomagenesis, and EBV-associated pathogenesis. This host background is particularly valuable for examining how mitochondrial perturbations intersect with oncogenic metabolism in B-cell contexts.
CMC2 encodes a mitochondrial intermembrane space protein that acts as a chaperone for the assembly of cytochrome c oxidase (Complex IV), the terminal enzyme of the mitochondrial electron transport chain. It is critically involved in the copper insertion step required for COX holoenzyme formation. The CMC2 protein interacts directly with copper chaperones COX11, SCO1, and SCO2, as well as with the mitochondrially encoded core subunits MT-CO1 and MT-CO2. Its expression is under the transcriptional regulation of PGC-1?? and NRF1, which are central nodes in mitochondrial biogenesis signaling. Disruption of CMC2 consequently leads to defective COX assembly, diminished electron transport through complexes I?CV, reduced mitochondrial respiration, and decreased ATP synthesis, thereby impairing oxidative phosphorylation.
In the Raji B-cell malignancy context, CMC2 knockout provides a highly relevant platform to probe the interplay between mitochondrial energy metabolism and the Warburg effect. Burkitt lymphoma cells typically exhibit high glycolytic rates, but residual oxidative phosphorylation remains essential under certain conditions; loss of CMC2 enables dissection of this dependency. Furthermore, this model mimics molecular features of mitochondrial complex IV deficiency disorders, such as Leigh syndrome and mitochondrial encephalomyopathy, offering a tractable system to study copper-dependent COX biogenesis defects in a transformed B-cell background with defined surface markers.
Key research applications include mitochondrial dysfunction studies, oxidative phosphorylation analysis, and drug screening for COX deficiencies. Researchers can employ a combination of functional assays: Western blotting for COX subunits and RT-qPCR for MT-CO1 and MT-CO2 to assess complex IV expression; Seahorse respirometry to quantify oxygen consumption rates; ATP luminescence assays to gauge energy output; TMRE/TMRM staining for mitochondrial membrane potential; ROS detection for oxidative stress evaluation; and direct complex IV enzyme activity measurements. These approaches make the product suitable for cancer metabolism research, mitochondrial disease modeling, and metabolic inhibitor testing. For further technical details or custom requests, please contact Ascent Research.
