The KEAP1 Knockout 786-O Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal cell population featuring targeted disruption of KEAP1 in the 786-O human renal cell adenocarcinoma cell line. This pooled knockout format provides a heterogeneous loss-of-function model without the need for single-cell clone isolation, facilitating robust functional genomics studies. The polyclonal nature reduces potential clonal artifacts and is well-suited for investigating KEAP1-dependent signaling, drug response profiling, and high-throughput screening applications.
The 786-O host cell line is a classic model of clear cell renal cell carcinoma (ccRCC) derived from a primary tumor. It harbors a biallelic VHL mutation, leading to constitutive stabilization of hypoxia-inducible factors (HIFs) and partial activation of HIF-driven transcriptional programs. This VHL-deficient background offers a clinically relevant platform to explore crosstalk between the hypoxic response and the KEAP1?CNRF2 axis, as both pathways are frequently dysregulated in ccRCC and contribute to tumor progression and therapy resistance.
KEAP1 acts as a substrate adaptor for the CUL3?CRBX1 E3 ubiquitin ligase complex, which ubiquitinates NRF2 (NFE2L2) to promote its proteasomal degradation under homeostatic conditions, thereby suppressing the antioxidant response. Oxidative or electrophilic stress triggers modification of KEAP1 cysteine residues by agents such as sulforaphane or reactive oxygen species, or PKC-mediated phosphorylation, disrupting the KEAP1?CNRF2 interaction. Stabilized NRF2 translocates to the nucleus and induces expression of cytoprotective genes??including NQO1, HMOX1, GCLM, and TXNRD1??via the antioxidant response element. KEAP1 also engages with p62/SQSTM1, PGAM5, and IKK??, linking oxidative stress sensing to autophagy, mitochondrial function, and inflammatory signaling.
In 786-O cells, inactivation of KEAP1 is expected to result in NRF2 hyperactivation, driving constitutive upregulation of antioxidant and detoxification enzymes. This may confer enhanced resistance to oxidative stress and chemotherapeutic agents, facilitating tumor cell survival and proliferation. Given the VHL-mutant background, dual dysregulation of HIF and NRF2 pathways could synergistically promote metabolic reprogramming and redox balance, underscoring the significance of this knockout model for studying aggressive ccRCC phenotypes and testing NRF2-targeted interventions.
This polyclonal knockout cell product is designed for applications such as dissecting NRF2 pathway hyperactivation, oxidative stress response mechanisms, and drug resistance in renal cell carcinoma. It supports screening of NRF2 inhibitors and functional assessment using assays like Western blotting for KEAP1 and NRF2, quantitative PCR for NRF2 targets (NQO1, HMOX1), NRF2 luciferase reporter systems, NRF2 nuclear translocation immunofluorescence, and cell viability under oxidative challenge. The model is a valuable tool for cancer biology and drug discovery. For further details or technical assistance, please contact Ascent Research.
