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Human MDM2 Knockout Cell Line-HEK293T

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
Description This cell is a stable cell line with a homozygous knockout of human MDM2 using CRISPR/Cas9.
Target Gene MDM2
Host Cell HEK293T
Host Cell Species Homo sapiens (Human)
Size Form 1 vial (>10^6 cell/vial)
Shipping Dry ice package
Storage Liquid nirtogen
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

Publications

Q & A

Customer Reviews

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.
Customer Q&As
What is the recommended growth medium? Does it require antibiotic selection?

A: DMEM supplemented with 10% fetal bovine serum.
It is not required to add the selection antibiotics when culturing the KO cells.

How is the knockout cell line validated?

A: The knockout cell product is validated by PCR amplification and Sanger Sequencing to confirm the mutation at the genomic level. Please find the detailed mutation info in the datasheet.

Is the product a single clonal cell or mixed cell pool?

A: Single clonal cell.

Can I confirm gene knockout by RT-qPCR?

A: No. This knockout cell product is generated using the CRISPR/Cas9 system to induce small insertions or deletions (indels) resulting in frameshift mutations. Although these frameshift mutations typically disrupt the coding gene, there is a possibility that the non-functional transcript may still be transcribed. Consequently, this could potentially yield misleading results when analyzed by RT-qPCR.

How can I store the cell product?

A: The cell line should be stored in liquid nitrogen for long-term preservation.

Is it possible to get multiple knockout clones for my GOI?

A: For most cases, we often keep at least 2 clones with different frameshift mutations. Please feel free to contact us to check if there are additional available clones.

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Customer Reviews
Excellent product!

This Human MDM2 Knockout Cell Line is robust and easy to maintain. It has provided us with reliable data, making it an invaluable tool in our laboratory.

United Kingdom

08/25/2024

Time-Saving

The knockout efficiency is outstanding, and the cells exhibit excellent growth and viability. This cell line has significantly accelerated our experimental timelines.

Canada

04/17/2022

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