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PKD1/PKD2 Double Knockout Cell Line-HEK293

PKD1/PKD2 Double Knockout Cell Line-HEK293

Cat.No. :  CSC-RT2612

Host Cell:  HEK293 Target Gene:  PKD1/PKD2

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

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Cell Line Information

Cell Culture Information

Safety and Packaging

Cat. No. CSC-RT2612
Description This cell is a stable cell line with a homozygous double knockout of human PKD1 and PKD2 using CRISPR/Cas9.
Target Gene PKD1/PKD2
Host Cell HEK293
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

Polycystic kidney disease 1 (PKD1) and polycystic kidney disease 2 (PKD2) are key genes in the development of autosomal dominant polycystic kidney disease (ADPKD), one of the most common inherited kidney diseases. The disease is characterized by the development of large, fluid-filled cysts in the kidneys, which can cause enlargement of these organs and eventually lead to kidney failure. PKD1 is located on chromosome 16 (16p13.3) and is responsible for encoding the protein polycystin-1. Polycystin-1 is an integral membrane protein composed of 4302 amino acids that is thought to play an important role in cell-cell and cell-matrix interactions. It is primarily localized in renal tubular epithelial cells and certain tissues outside the kidney, such as the heart and liver. Polycystin-1 is involved in a variety of cellular processes, including cell proliferation, differentiation, and apoptosis. Dysfunctional polycystin-1 due to PKD1 mutations disrupts these processes, leading to the formation and enlargement of kidney cysts. PKD2, on the other hand, is located on chromosome 4 (4q21) and encodes the protein polycystin-2. Unlike polycystin-1, polycystin-2 is a calcium-permeable, non-selective cation channel consisting of 968 amino acids. This protein is also expressed in the kidney tubules but is more widely distributed in various epithelial and muscle tissues. Polycystin-2 interacts with polycystin-1 to form a complex that regulates intracellular calcium levels, which is essential for a variety of cellular functions, including signaling pathways that control cell growth and division. PKD2 mutations disrupt the function of the polycystin-2 protein, further impairing calcium signaling and leading to cyst formation.
PKD1/PKD2 double knockout (KO) cell lines using HEK293 cells provide a powerful tool for scientific research and drug development. The following are the main applications of PKD1/PKD2 double knockout (KO) cell lines: Polycystic kidney disease (PKD) research: These cell lines are essential for studying autosomal dominant polycystic kidney disease (ADPKD) and help understand the molecular mechanisms of the disease, which involves mutations in the PKD1 and PKD2 genes. Drug screening and development: Double KO cell lines can be used to identify compounds that can alleviate or reverse the effects of PKD1 and PKD2 gene deletion, thereby providing potential therapeutic agents. Gene function analysis: By analyzing cellular changes upon PKD1 and PKD2 deletion, researchers can elucidate the role of these genes in cellular processes such as signal transduction, cilia formation, and ion transport. Genetic interaction studies: This cell line can be used to study genetic interactions and compensation mechanisms between PKD1, PKD2, and other genes, thereby revealing new pathways and biological processes affected by these genes.
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
User-Friendly

The PKD1/PKD2 Double Knockout Cell Line-HEK293 arrived well-packaged and was easy to thaw and culture. The instructions provided were clear and concise, allowing even my junior lab members to handle them with ease.

United States

02/22/2020

Worked very well

I’ve been using the Creative Biogene's PKD1/PKD2 Double Knockout Cell Line-HEK293 for my research on polycystic kidney disease and have found the results to be exceptionally consistent. The knockout efficiency is impressive, which has greatly enhanced the reliability of my experimental data.

United Kingdom

11/26/2022

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