Cat. No. ARG0086
KEAP1 Knockout A-549 is a human CRISPR/Cas9-edited lung adenocarcinoma epithelial cell line with disruption of the KEAP1 gene in the widely used A-549 NSCLC background. KEAP1 normally recruits NRF2/NFE2L2 to the CUL3-RBX1 ubiquitin ligase complex for proteasomal degradation; knockout therefore provides a relevant model for studying NRF2 stabilization, antioxidant response signaling, xenobiotic detoxification, glutathione metabolism, ferroptosis regulation, and drug resistance mechanisms. This model is well suited for redox biology, lung cancer research, western blotting, RT-qPCR, RNA-seq, ARE reporter assays, ROS measurements, metabolic assays, and drug sensitivity studies.
| Host Cell | A-549 |
| Morphology | Epithelial-like |
| Age | 58 years |
| Sex of Donor | Male |
| Gene Name | Keap1 |
| Gene Identifier | NCBI Gene ID 9817 |
| Temperature | 37°C |
| Atmosphere | 5% CO₂ |
| Sterility testing | Daily monitoring confirms that the cells are free from bacterial, yeast, and fungal contamination. |
| Mycoplasma testing | Negative for mycoplasma through PCR analysis |
| Pathogens | Cells tested negative for HIV-1, HBV, and HCV. |
Intended Use: This product is intended for laboratory in vitro use only. lt is not intended for diagnostic, therapeutic, or clinical applications.
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This product is provided "AS IS". For Research Use Only. Not for human or animal therapeutic use.
The KEAP1 Knockout A-549 Cell Line is a human CRISPR/Cas9-engineered cell model in which the KEAP1 gene has been disrupted to eliminate functional KEAP1 expression. This stable in vitro knockout system is generated in the A-549 background, a human lung adenocarcinoma epithelial cell line, and is intended for mechanistic studies of redox-regulated signaling, cancer cell adaptation, and therapeutic response. As a gene-edited derivative of a widely used pulmonary tumor model, it provides a defined platform for examining the cellular consequences of KEAP1 loss in a lung epithelial-like cancer context.
A-549 cells are derived from human non-small cell lung cancer and exhibit epithelial characteristics relevant to lung adenocarcinoma biology. This host line is extensively used in studies of pulmonary cell biology, xenobiotic metabolism, oxidative stress signaling, and drug sensitivity, in part because it recapitulates key features of alveolar epithelial-like tumor cells and supports robust molecular and pharmacologic interrogation. The model is therefore suitable for investigating disease-relevant processes in lung cancer, as well as broader questions in toxicology, redox homeostasis, and adaptation to environmental or therapeutic stress.
KEAP1 is a redox-sensitive substrate adaptor that forms a functional complex with CUL3 and RBX1 to promote constitutive ubiquitination and proteasomal degradation of NFE2L2/NRF2 under basal conditions. Its activity is regulated by reactive oxygen species, electrophiles, lipid peroxidation products, and NRF2-inducing oxidants, and is further modulated by interacting factors including SQSTM1/p62, PGAM5, PALB2, IKBKB, and HSP90. Disruption of KEAP1 relieves repression of NRF2, resulting in NRF2 stabilization, nuclear accumulation, and transcriptional activation of ARE-containing target genes such as HMOX1, NQO1, GCLC, GCLM, SLC7A11, TXNRD1, GSTP1, AKR1C1, and FTH1. Through these outputs, the KEAP1-NRF2 axis acts upstream of antioxidant defense, glutathione metabolism, xenobiotic detoxification, reactive oxygen species homeostasis, ferroptosis regulation, and metabolic reprogramming.
In the A-549 background, KEAP1 loss is particularly relevant for studying lung adenocarcinoma-associated redox adaptation, treatment resistance, and pathway dependency. Because A-549 cells are commonly used to model oxidative stress responses and pulmonary xenobiotic biology, KEAP1 knockout provides a useful system for defining how sustained NRF2 activation reshapes transcriptional programs, stress tolerance, and sensitivity to chemotherapeutic agents, radiation, or ferroptosis-inducing conditions in non-small cell lung cancer.
This cell line can be applied in western blot and immunofluorescence workflows to assess NRF2 stabilization and nuclear localization; RT-qPCR, RNA-seq, and ARE reporter assays to quantify activation of HMOX1, NQO1, GCLC, SLC7A11, and related target genes; and co-immunoprecipitation or ubiquitination assays to study disruption of KEAP1-CUL3-RBX1-mediated substrate regulation. It is also suitable for ROS measurements, glutathione assays, flow cytometry, metabolic profiling, colony formation assays, ferroptosis assays, and drug sensitivity studies designed to interrogate chemoresistance, redox liabilities, and synthetic lethal interactions. Researchers may contact Ascent Research for additional technical information, product details, or related gene-edited cell models.
