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Human MC4R Knockout Cell Line-HEK293T

Human MC4R Knockout Cell Line-HEK293T

Cat.No. :  CSC-RT2778

Host Cell:  HEK293T Target Gene:  MC4R

Size:  1x10^6 cells/vial, 1mL Validation:  Sequencing

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Cell Line Information

Cell Culture Information

Safety and Packaging

Cat. No. CSC-RT2778
Description This cell is a stable cell line with a homozygous knockout of human MC4R using CRISPR/Cas9.
Target Gene MC4R
Host Cell HEK293T
Host Cell Species Homo sapiens (Human)
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.
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

The melanocortin 4 receptor (MC4R) gene is located on human chromosome 18 and encodes the MC4R protein, a G protein-coupled receptor that is primarily expressed in the brain, particularly in regions such as the hypothalamus that are involved in appetite and energy balance. MC4R is a key component of the central melanocortin pathway, influencing energy expenditure and food intake. Activation of MC4R by its natural ligands, such as α-melanocyte stimulating hormone (α-MSH), results in decreased food intake and increased energy expenditure, thus promoting weight loss. Conversely, inhibition or dysfunction of MC4R leads to overeating (excessive eating) and decreased energy expenditure, resulting in obesity. MC4R gene mutations are one of the most common genetic causes of severe obesity. Both heterozygous and homozygous mutations have been found, with the latter generally resulting in a more severe phenotype. These mutations disrupt the normal function of the receptor, resulting in impaired melanocortin pathway signaling. As a result, individuals with MC4R dysfunction may be more susceptible to weight gain and may find weight control more challenging. In addition to its role in obesity, MC4R is involved in other physiological processes, including regulation of glucose metabolism, sexual function, and cardiovascular maintenance. Studies have shown that MC4R mutations may also be associated with other metabolic disorders, such as insulin resistance and type 2 diabetes.
The human MC4R knockout cell line - HEK293T is a valuable tool in scientific research and drug development. The following are the main applications of this cell line: Obesity and Metabolic Disorder Research: The MC4R receptor plays an important role in regulating body weight and energy homeostasis. Knockout of the MC4R gene in HEK293T cells helps researchers understand the molecular mechanisms behind obesity and metabolic disorders. Drug Screening and Development: HEK293T cells lacking MC4R can be used in high-throughput screening assays to identify potential agonists or antagonists against this receptor. Such compounds may lead to new treatments for obesity and related metabolic diseases. Signal Transduction Studies: These cells provide a controlled environment to study downstream signaling pathways activated by MC4R. This is essential for elucidating the role of the receptor in various physiological processes and identifying nodes that may be targeted by therapeutic agents. Genetic Studies: By comparing the behavior of MC4R knockout cells with wild-type cells, researchers can study the functions and interactions of genes. This comparative approach can reveal insights into gene networks and molecular players involved in energy balance. Pharmacological Studies: MC4R knockout HEK293T cells are essential for studying the pharmacodynamics and pharmacokinetics of drugs targeting this receptor. These studies help understand drug-receptor interactions, drug efficacy and potential side effects, and guide the dosing and administration strategies of new therapies.
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
Worked very well

The Human MC4R Knockout Cell Line-HEK293T has demonstrated exceptional quality and reliability in our obesity research. The knockout efficiency is impressive, and the cells have shown consistent results across multiple experiments.

Germany

10/07/2021

Helpful

The knockout model accurately represents the loss-of-function studies we aimed to perform, thus enabling rapid discovery and validation of potential therapeutic targets.

French

05/25/2020

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