Human MDM2 Knockout Cell Line-HEK293T
Cat.No. : CSC-RT2718
Host Cell: HEK293T Target Gene: MDM2
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT2718 |
| Cell Line Information | This cell is a stable cell line with a homozygous knockout of human MDM2 using CRISPR/Cas9. |
| Target Gene | MDM2 |
| Host Cell | HEK293T |
| Size Form | 1 vial (>10^6 cell/vial) |
| Shipping | Dry ice package |
| Storage | Liquid nirtogen |
| Species | Human |
| 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 1:3-1:6. |
| 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 |
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Background
Applications
MDM2, also known as E3 ubiquitin protein ligase Mdm2, is a key gene in humans that plays an important role in regulating the p53 tumor suppressor protein. The gene was originally discovered in mice and is known as "murine double differential 2" (mdm2). MDM2 is both an oncogene and an E3 ubiquitin ligase. As an oncogene, MDM2 promotes tumor formation and growth by inhibiting p53, which is responsible for initiating cell cycle arrest and apoptosis in response to stress signals.
Mdm2 inhibits the transcriptional activity of p53 by binding to the N-terminal transactivation domain of p53, thereby preventing its ability to activate target genes. In addition, MDM2 mediates the ubiquitination and subsequent proteasomal degradation of p53, resulting in a decrease in intracellular p53 levels. This feedback loop ensures that p53 levels remain low under normal conditions, thereby preventing unnecessary cell cycle arrest or apoptosis.
The role of MDM2 in tumorigenesis is well established, with elevated levels of Mdm2 observed in various human cancers, including soft tissue sarcomas, osteosarcomas, and breast tumors. Overexpression of MDM2 can promote oncogenic transformation in concert with other oncogenes, such as Ras, leading to tumor formation.
1. Cancer Research: MDM2 is a well-known negative regulator of the p53 tumor suppressor. Knockout of MDM2 in HEK293T cells allows researchers to study the role of p53 in tumorigenesis, enabling the identification of novel therapeutic targets and the development of p53-based cancer therapies.
2. Drug Screening: MDM2 knockout cell lines can be used for high-throughput screening of potential anticancer drugs. Drugs targeting the p53-MDM2 interaction can be evaluated for efficacy and toxicity, providing valuable data for the drug development pipeline.
3. Genomic Studies: Researchers can use MDM2 knockout cell lines to study genetic and epigenetic changes that occur as a result of loss of MDM2 function. This can provide a better understanding of gene regulatory networks and molecular pathways involved in cell cycle regulation and apoptosis.
4. Protein Interaction Studies: The effects of MDM2 on various p53-dependent and -independent pathways can be explored by studying protein-protein interactions in knockout cell lines. This helps identify new interactions and understand the broader impact of MDM2 function beyond p53 regulation.
5. Cell Cycle Analysis: By knocking out MDM2 in HEK293T cells, researchers can analyze changes in the cell cycle, specifically the mechanisms that control cell arrest, DNA repair, and apoptosis. This can elucidate the mechanisms by which MDM2 affects cell proliferation and survival.
For research use only. Not intended for any clinical use.
