The CCS Knockout AGS Polyclonal Cells constitute a CRISPR/Cas9-mediated gene-disrupted heterogeneous cell population derived from the human gastric adenocarcinoma line AGS. This polyclonal knockout pool targets the CCS gene, which encodes the copper chaperone for superoxide dismutase 1 (SOD1), and is supplied as an unsorted mix of edited cells. The product is designed to enable loss-of-function studies of CCS-dependent processes in a gastric epithelial context, without the need for single-cell cloning. It provides a versatile experimental system for interrogating copper chaperone biology and redox regulation in cancer cells.
AGS cells are an adherent epithelial cell line originally established from a human gastric adenocarcinoma and are widely employed to model gastric cancer pathobiology. These cells harbor an activating KRAS mutation while maintaining wild-type p53 status, offering a genetically defined background that mimics key oncogenic features of gastric adenocarcinoma. The epithelial nature of AGS cells supports physiologically relevant investigations of gastric mucosal cell functions, including those governing trace metal metabolism and oxidative stress responses. Their robust growth and ease of manipulation render them suitable for a broad spectrum of molecular and pharmacological assays.
CCS functions as a dedicated copper chaperone that specifically delivers copper ions to SOD1, an indispensable step for the enzymatic maturation and superoxide dismutase activity of SOD1, which catalyzes the dismutation of toxic superoxide radicals into oxygen and hydrogen peroxide. CCS activity is regulated by the metal-responsive transcription factor MTF1, cellular copper availability, and oxidative stress cues, placing it at the nexus of copper homeostasis and antioxidant defense. The copper delivery network additionally involves the high-affinity copper importer CTR1, the cytosolic copper carrier ATOX1, and reduced glutathione, all of which cooperate to ensure precise copper incorporation into SOD1. Disruption of CCS expression is predicted to impair SOD1 activation, leading to elevated intracellular reactive oxygen species (ROS) levels, altered glutathione redox cycling, and potential perturbation of downstream signaling pathways sensitive to redox balance.
In the AGS gastric cancer model, which carries an oncogenic KRAS mutation known to drive ROS production and adaptive antioxidant programs, CCS knockout introduces a unique paradigm for dissecting the interplay between copper-dependent SOD1 activation and redox homeostasis in tumorigenesis. KRAS-mutant carcinomas often exhibit heightened reliance on antioxidant defenses to mitigate oxidative stress; therefore, loss of CCS may expose vulnerabilities related to ROS detoxification or trigger compensatory mechanisms in copper handling and alternative antioxidant pathways. Furthermore, studying CCS disruption in a gastric epithelial background permits exploration of copper biology in a cancer-relevant context, where copper levels have been implicated in angiogenesis, proliferation, and metastasis. This model facilitates the dissection of CCS as a potential molecular switch linking copper metabolism to oncogenic ROS signaling.
This CCS knockout polyclonal AGS population supports a range of experimental applications, including the analysis of copper chaperone-mediated SOD1 activation by immunoblotting for CCS and SOD1 protein levels, superoxide dismutase activity assays, and intracellular ROS detection using fluorescent reporters such as H2DCFDA. Copper quantification and RT-qPCR for CCS and SOD1 transcripts further enable detailed examination of copper homeostasis and transcriptional responses. The model is well-suited for drug sensitivity screens with copper chelators like tetrathiomolybdate or ROS-inducing agents such as hydrogen peroxide, allowing assessment of redox-dependent cell viability and proliferation. It also provides a platform to investigate crosstalk between KRAS-driven oncogenic signals and copper-regulated oxidative stress pathways. For additional information or technical inquiries, please contact Ascent Research.
