Human MDM2 Knockout Cell Line-HeLa
Cat.No. : CSC-RT0516
Host Cell: HeLa Target Gene: MDM2
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
Inquire for Price
Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT0516 |
| Cell Line Information | A stable cell line with a homozygous knockout of human MDM2 using CRISPR/Cas9. |
| Target Gene | MDM2 |
| Host Cell | HeLa |
| Shipping | 10^6 cells/tube |
| Storage | Liquid nitrogen |
| Species | Human |
| Gene ID | 4193 |
| 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. |
| Media Type | Cells were cultured in DMEM supplemented with 10% fetal bovine serum. |
| Growth Properties | Cells are cultured as a monolayer at 37°C in a humidified atmosphere with 5% CO2. Split at 80-90% confluence, approximately 1:4-1:6. |
| Freeze Medium | Complete medium supplemented with 10% (v/v) DMSO |
| 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 |
Inquiry
Background
Case Study
Applications
MDM2 (Mouse Double Minute 2 homolog) is a key protein that regulates the p53 tumor suppressor pathway and plays a critical role in cell cycle control and apoptosis. MDM2 is an E3 ubiquitin ligase, which means that it tags specific proteins with ubiquitin to target them for degradation via the proteasome, an important process for maintaining cellular homeostasis. Under normal conditions, MDM2 binds to p53, inhibiting its activity and leading to its degradation. This interaction is critical because p53 is a potent transcription factor that promotes the expression of genes involved in cell cycle arrest and apoptosis in response to cellular stress and DNA damage. By regulating p53 levels, MDM2 ensures that p53 activity is kept in check, preventing unnecessary cell cycle arrest or cell death.
However, in many cancers, this balance is disrupted. Overexpression of MDM2, often due to gene amplification or other mechanisms, results in reduced p53 activity, allowing cells with damaged DNA to proliferate uncontrollably and escape apoptosis. Studies of small molecule inhibitors of MDM2 designed to disrupt its interaction with p53 have shown promise in preclinical models and clinical trials. These inhibitors are intended to reactivate p53 function, thereby restoring its tumor suppressor activity and providing a new strategy for cancer treatment.
MDM2 (murine double minute 2) is a key inhibitor of p53-mediated tumor suppressor functions, including cellular senescence. Werner syndrome (WS) is caused by mutations in the WRN gene and is an autosomal recessive disorder. Loss of WRN function induces senescence in human cancer cells. Here, the researchers show that MDM2 acts as an E3 ligase for the WRN protein. MDM2 interacts with WRN both in vivo and in vitro. MDM2 induces WRN ubiquitination and significantly downregulates the levels of WRN protein in human cells. During the DNA damage response, WRN is translocated to the nucleoplasm to promote its DNA repair function. However, it is degraded by the MDM2-mediated ubiquitination pathway. Furthermore, the senescence phenotype induced by DNA damaging agents, such as etoposide, is at least partially mediated by MDM2-dependent WRN degradation, as ectopic expression of WRN can significantly attenuate this degradation. These results demonstrate that MDM2 plays a critical role in regulating WRN function through ubiquitin-dependent degradation and reveal an unexpected role for MDM2 in promoting cellular senescence in a p53-independent manner.
Here, the researchers performed cycloheximide (CHX) chase experiments to examine whether MDM2 could regulate the half-life of WRN protein by transfecting HCT116 cells with a combination of plasmids producing MYC-WRN and MDM2. When MDM2 was co-expressed, the half-life of WRN decreased dramatically (Figure 1a). In addition, the half-life of WRN was increased in the MDM2 knockout cell line compared to the wild-type cell line (Figure 1b).
Figure 1. a When MDM2 was coexpressed, the half-life of WRN decreased dramatically. B WRN half-life was increased in MDM2 knockout cell lines compared with that in wild-type cell lines. (Liu, Boya, et al. 2019)
The Human MDM2 Knockout Cell Line-HeLa is a powerful research tool with multiple applications in the fields of cancer biology, molecular genetics, and drug development. Here are some key applications:
1. Cancer Research: MDM2 is a key regulator of the p53 tumor suppressor protein. By using HeLa cells with MDM2 knockout, researchers can study the role of MDM2 in tumor progression and p53-mediated cellular processes.
2. p53 Pathway Studies: Since MDM2 negatively regulates p53, MDM2 knockout HeLa cells provide an excellent model for studying the activation and function of the p53 pathway. Researchers can explore how the loss of MDM2 affects p53 stability, transcriptional activity, and its ability to induce cell cycle arrest or apoptosis in response to DNA damage or other stress signals.
3. Drug Screening and Development: The Human MDM2 Knockout Cell Line-HeLa can be used to screen for compounds that activate or inhibit the p53 pathway.
4. Genetic and Molecular Studies: These cells can be used to dissect the genetic and molecular interactions between MDM2, p53, and other signaling pathways.
5. Apoptosis and Cell Cycle Regulation Studies: Researchers can explore how loss of MDM2 affects apoptosis and cell cycle regulation in HeLa cells. This is particularly important for understanding the checks and balances of cell division and programmed cell death processes, which are often dysregulated in cancer.
For research use only. Not intended for any clinical use.
