Human KEAP1 Knockout Cell Line-A549
Cat.No. : CSC-RT2794
Host Cell: A549 Target Gene: KEAP1
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT2794 |
| Cell Line Information | This cell is a stable cell line with a homozygous knockout of human KEAP1 using CRISPR/Cas9. |
| Target Gene | KEAP1 |
| Host Cell | A549 |
| Size Form | 1 vial (>10^6 cell/vial) |
| Shipping | Dry ice package |
| Storage | Liquid Nitrogen |
| Revival | Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media. |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
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Background
Applications
KEAP1 (Kelch-like ECH-associated protein 1) is a key gene involved in the cellular defense mechanism against oxidative stress. KEAP1 functions primarily as a negative regulator of NRF2. Under basal conditions, KEAP1 binds to NRF2, promoting its ubiquitination and subsequent proteasomal degradation. This interaction ensures that NRF2 levels remain low under normal physiological conditions. However, under the action of oxidative stress or electrophilic compounds, KEAP1 undergoes conformational changes that prevent its binding to NRF2, allowing NRF2 to accumulate and translocate into the nucleus. Once in the nucleus, NRF2 activates the expression of target genes involved in antioxidant defense, detoxification, and cell survival.
Mutations and dysregulation of the KEAP1 gene are associated with various diseases, especially cancer. Loss-of-function mutations in KEAP1 can lead to uncontrolled activation of NRF2, resulting in excessive cell proliferation and survival. Such mutations are common in non-small cell lung cancer (NSCLC), liver cancer, and several other malignancies. Increased NRF2 activity provides cancer cells with a survival advantage, making them more resistant to chemotherapy and radiation therapy, posing a major challenge to cancer treatment. In addition, KEAP1 mutations have also been associated with neurodegenerative diseases, in which impaired oxidative stress responses may lead to neuronal damage.
Applications of Human KEAP1 Knockout Cell Line (A549)
1. Cancer Research: The KEAP1 knockout cell line (A549) is a valuable tool for studying the mechanisms of lung cancer. By knocking out the KEAP1 gene, researchers can investigate how the cellular pathways change, leading to cancer progression. This can lead to the identification of novel therapeutic targets specific to KEAP1 mutations or deletions.
2. Oxidative Stress Studies: KEAP1 is a crucial regulator of the antioxidant response. In KEAP1 knockout cells, the NRF2 pathway is constitutively activated. This makes this cell line an excellent model for studying oxidative stress and the cellular response to various oxidative agents, potentially leading to new insights into oxidative stress-related diseases.
3. Drug Screening: This cell line can be used in high-throughput drug screening to identify compounds that selectively target KEAP1-deficient cells. Such compounds can be promising candidates for the development of new cancer treatments, particularly for patients with KEAP1 mutations.
4. Pathway Dissection: With KEAP1 knocked out, it becomes possible to dissect the downstream effects and interactions within various cellular signaling pathways. This can shed light on how KEAP1 interacts with other proteins and pathways, contributing to a more comprehensive understanding of cellular homeostasis and response to stress.
5. Biomarker Discovery: By comparing the KEAP1 knockout cell line with the wild-type A549 cells, scientists can identify potential biomarkers that are specifically associated with the KEAP1-NRF2 pathway.
For research use only. Not intended for any clinical use.
