The CCS Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of K-562 human chronic myelogenous leukemia (CML) cells carrying targeted disruption of the CCS gene. This polyclonal knockout model provides a loss-of-function tool for investigating copper chaperone CCS-dependent processes without clonal isolation, preserving population-level genetic heterogeneity. The product is designed for researchers studying copper homeostasis, oxidative stress, and SOD1 activation in a leukemic background.
K-562 is a suspension cell line derived from a 53-year-old female CML patient in blast crisis. The cells harbor the Philadelphia chromosome, expressing the BCR-ABL fusion oncoprotein, which drives constitutive tyrosine kinase signaling. K-562 cells exhibit erythroleukemia features and serve as a widely used model for myeloid leukemia, particularly suited for examining oncogene-driven redox perturbations and stress responses.
CCS encodes a copper chaperone that shuttles copper from the plasma membrane importer CTR1 (SLC31A1) to apo-SOD1, activating superoxide dismutase for superoxide radical detoxification. Downstream, hydrogen peroxide produced by SOD1 is metabolized by catalase and glutathione peroxidase. CCS directly interacts with SOD1 and is functionally connected to ATOX1 and the copper-transporting ATPase ATP7A. Its expression is regulated by metal-responsive transcription factor MTF-1, copper availability, and HIF-1. Knockout of CCS blocks copper delivery to SOD1, impairing enzymatic activity and leading to accumulation of superoxide radicals, thereby disrupting cellular redox homeostasis. This mechanism is highly relevant to neurodegenerative diseases like amyotrophic lateral sclerosis (ALS) and copper dysregulation disorders.
In K-562 leukemia cells, CCS knockout exacerbates the endogenous oxidative stress driven by BCR-ABL signaling. Leukemic cells maintain high ROS levels and depend on robust antioxidant defenses. Disabling CCS-mediated SOD1 activation sensitizes these cells to oxidative damage, offering a platform to study copper-dependent survival mechanisms, redox vulnerability in CML, and potential synergy with BCR-ABL inhibitors or pro-oxidant therapies.
This knockout model supports diverse applications including copper uptake assays with 64Cu, western blotting and enzymatic assays for SOD1 activation, flow cytometry for ROS detection (DCFDA), RT-qPCR for oxidative stress gene profiling, and immunofluorescence to examine SOD1 localization. Functional studies may involve cell viability and apoptosis assays under hydrogen peroxide stress. It is suitable for drug screening targeting copper chaperones or antioxidant pathways. For additional details, please contact Ascent Research.
