The KEAP1 Knockout A2780 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A2780 human ovarian carcinoma cell line. This product provides a loss-of-function model for the KEAP1 gene, generated through CRISPR/Cas9-mediated gene disruption. The polyclonal nature of these cells preserves genetic heterogeneity, making them suitable for studies requiring mixed knockout populations rather than clonal isolates. This KEAP1 knockout model is an essential tool for investigating the KEAP1-NRF2 signaling axis in ovarian cancer biology.
The A2780 cell line is a well-established human ovarian epithelial carcinoma model isolated from an untreated patient. It is widely employed in research on cisplatin resistance, a major challenge in ovarian cancer treatment. A2780 cells retain key oncogenic signaling properties, and their epithelial origin makes them a relevant system for studying tumor cell responses to chemotherapeutic stress. Their use in combination with KEAP1 knockout provides a physiologically contextualized platform for dissecting resistance mechanisms and redox regulation.
KEAP1 acts as a critical sensor of oxidative and electrophilic stress by functioning as an adaptor for the CUL3-RBX1 E3 ubiquitin ligase complex, which targets NRF2 for constitutive proteasomal degradation. Under basal conditions, KEAP1 binds NRF2 and facilitates its ubiquitination. Oxidative or electrophilic stimuli, including reactive oxygen species and electrophilic compounds, modify KEAP1 cysteine residues, disrupting NRF2 ubiquitination and enabling NRF2 stabilization. Accumulated NRF2 translocates to the nucleus, where it binds antioxidant response elements and drives the expression of detoxifying and antioxidant genes such as NQO1, HMOX1, GCLC, GCLM, and TXNRD1. KEAP1 also interacts with CUL3, RBX1, p62/SQSTM1, PGAM5, and IKK??, integrating diverse stress signals. In the knockout model, loss of KEAP1 results in constitutive NRF2 stabilization and sustained activation of cytoprotective programs, offering a system to dissect persistent NRF2 signaling.
In the A2780 ovarian carcinoma background, KEAP1 disruption profoundly alters cellular redox balance and drug responsiveness. Constitutive NRF2 activity elevates antioxidant and detoxification capacity, reducing sensitivity to cisplatin. This is especially relevant since cisplatin resistance often involves upregulated glutathione metabolism and redox buffering. Thus, the KEAP1 knockout A2780 cells serve as a model to examine NRF2-driven drug susceptibility changes, proliferation under oxidative stress, and metabolic reprogramming, revealing potential therapeutic vulnerabilities.
These polyclonal knockout cells are suitable for a range of assays including western blotting for NRF2, NQO1, and HMOX1; RT-qPCR of antioxidant genes; glutathione quantification; and ROS measurement. They facilitate viability studies with cisplatin to assess chemoresistance and immunofluorescence for NRF2 localization. Researchers can also employ the cells in high-throughput screening for NRF2 modulators or metabolic studies. For further inquiries, please contact Ascent Research.
