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Human PKD2 Knockout Cell Line-HEK293

Human PKD2 Knockout Cell Line-HEK293

Cat.No. :  CSC-RT2611

Host Cell:  HEK293 Target Gene:  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-RT2611
Description This cell is a stable cell line with a homozygous knockout of human PKD2 using CRISPR/Cas9.
Target Gene 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

The PKD2 gene, also known as polycystic kidney disease 2, is an important gene that plays a key role in human physiology and pathology. The PKD2 gene is located on chromosome 4q22.1 and encodes a membrane protein called polycystin-2, which is involved in the formation and function of primary cilia and calcium signaling pathways. The main role of the PKD2 gene is closely related to its function in calcium ion transport and its interaction with other proteins such as polycystin-1 encoded by the PKD1 gene. Both polycystin-1 and polycystin-2 are essential for maintaining the structure and function of the renal tubules. PKD2 gene mutations are associated with autosomal dominant polycystic kidney disease (ADPKD), a common inherited disease characterized by the presence of numerous cysts in the kidneys. The disease can lead to progressive renal failure, hypertension, and other systemic complications. Although PKD2 mutations cause a smaller proportion of ADPKD cases compared to PKD1 mutations, the clinical manifestations of PKD2-related ADPKD tend to be milder and occur later. In addition to its role in ADPKD, studies suggest that the PKD2 gene may have other physiological functions and may be involved in other diseases involving ciliary function and calcium signaling.
PKD2 encodes polycystin-2, a key protein involved in kidney function and cellular processes. Creation of a PKD2 knockout model in HEK293 cells has helped to better understand the molecular mechanisms behind PKD and aid in drug discovery, gene function studies, and therapeutic development. Below are the main applications of the human PKD2 knockout cell line - HEK293. Polycystic Kidney Disease (PKD) Research: PKD2 knockout HEK293 cells provide a simplified in vitro model to study the pathophysiology of polycystic kidney disease. By examining the cellular and molecular consequences of PKD2 deficiency, researchers can identify disease mechanisms, progression, and key regulatory pathways. Functional Analysis of Polycystin-2: Loss of PKD2 in HEK293 cells highlights the role of polycystin-2 in cellular processes such as calcium signaling, cell proliferation, and apoptosis. This helps to study how polycystin-2 interacts with other proteins and regulates cellular homeostasis. Drug Screening and Development: Using PKD2 knockout HEK293 cells, researchers can screen for potential therapeutic compounds that can compensate for loss of polycystin-2 function or correct its downstream effects. This cell line provides a high-throughput system to test drug efficacy and toxicity, thereby accelerating the identification of promising drug candidates. Molecular and Cellular Pathway Elucidation: By comparing PKD2 knockout cells to wild-type HEK293 cells, researchers can identify alterations in cellular signaling pathways, gene expression profiles, and metabolic processes. This comparative analysis helps delineate pathways affected by PKD2 loss and may reveal new therapeutic targets.
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
Invaluable tool

The PKD2 knockout cell generation technique is a valuable tool for investigating the mechanisms involved in PKD pathogenesis. I recommend Creative Biogene'cell line.

United States

02/23/2023

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