The CCS Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited population of human NCI-H1299 lung adenocarcinoma cells carrying a disrupted CCS gene. This polyclonal knockout model enables loss-of-function studies of the copper chaperone CCS, facilitating investigation of copper-dependent antioxidant defense and oxidative stress pathways. The cells are provided as a heterogeneous polyclonal pool, ready for immediate use in functional assays.
The NCI-H1299 cell line originates from a lymph node metastasis of a non-small cell lung carcinoma and is p53-deficient. This p53-null background is characteristic of aggressive lung tumors and provides a relevant setting for examining cellular responses to oxidative and genotoxic stress, independent of canonical p53-mediated pathways. The metastatic origin of the cells further adds value for studying invasive cancer mechanisms.
CCS plays a central role in copper distribution by specifically donating copper to superoxide dismutase 1 (SOD1), the enzyme responsible for superoxide radical detoxification. CCS activity is modulated by upstream regulators including cellular copper levels, oxidative stress, and the metal-responsive transcription factor MTF1. Upon copper loading, CCS interacts directly with SOD1 and is thought to also associate with the copper transporter CTR1 and the apoptosis inhibitor XIAP. Disruption of CCS reduces SOD1 activity, causing superoxide accumulation that can activate redox-sensitive transcription factors NF-??B and AP-1, ultimately leading to oxidative damage of lipids, proteins, and DNA.
In the p53-deficient NCI-H1299 context, CCS knockout is particularly valuable for modeling oxidative stress vulnerabilities in lung cancer. The impairment of SOD1 function is expected to heighten baseline ROS levels and sensitize cells to oxidative insults, making this model suitable for studying tumor cell adaptation to redox imbalance. It also serves as a platform to dissect the interplay between copper homeostasis and antioxidant defenses in a metastatic cancer background.
These CCS knockout cells support a wide range of experimental approaches, including Western blotting for protein expression, SOD activity assays, and DCFDA-based ROS detection. They are applicable to copper uptake studies, clonogenic survival under oxidative stress, immunofluorescence, and quantitative gene expression analysis by RT-qPCR or RNA-seq. Research applications span investigation of SOD1 function in cancer, oxidative stress response mechanisms, copper metabolism, and screening of antioxidant or pro-oxidant therapeutic candidates. For additional technical inquiries, please contact Ascent Research.
