Human CFTR Knockout Cell Line-HEK293T
Cat.No. : CSC-RT2753
Host Cell: HEK293T Target Gene: CFTR
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT2753 |
| Cell Line Information | This cell is a stable cell line with a homozygous knockout of human CFTR using CRISPR/Cas9. |
| Target Gene | CFTR |
| Host Cell | HEK293T |
| Size Form | 1 vial (>10^6 cell/vial) |
| Shipping | Dry ice package |
| Storage | Liquid Nitrogen |
| 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
The cystic fibrosis transmembrane conductance regulator (CFTR) is a membrane protein and anion channel found in vertebrates and encoded by the CFTR gene. The CFTR gene belongs to the ATP-binding cassette (ABC) transporter family and encodes a protein that regulates the movement of chloride and bicarbonate ions across epithelial cell membranes. This regulation is essential for maintaining fluid balance in various organs such as the lungs and pancreas. The role of CFTR is not limited to the transport of chloride and bicarbonate ions. It is also involved in regulating other ion channels, affecting acid-base balance, and participating in cellular defense mechanisms. The gene is highly expressed in various tissues, including the respiratory tract, digestive system, and reproductive organs. Its complex structure with two transmembrane domains, two nucleotide binding domains, and a regulatory domain gives it a complex function that requires phosphorylation and ATP binding for activation.
Mutations in the CFTR gene disrupt its ion channel function, resulting in improper chloride and bicarbonate ion transport. Since its discovery, more than 700 CFTR mutations have been found, each resulting in a different expression and severity of the disease. The most common mutation, Delta F508, causes the CFTR protein to misfold and degrade before it reaches the cell membrane. As a result, the absence of a functional CFTR channel leads to the formation of thick mucus in organs such as the lungs and pancreas, which can cause recurrent lung infections, pancreatic insufficiency, malnutrition, and diabetes. Male patients may also be born with bilateral absence of the vas deferens, resulting in infertility.
1. Pathophysiological Studies of Cystic Fibrosis: CFTR knockout HEK293T cells provide a powerful model for studying the molecular and cellular mechanisms of cystic fibrosis. The lack of functional CFTR protein in these cells mimics the cellular environment of cystic fibrosis patients, allowing researchers to study disease pathophysiology in a controlled environment.
2. Drug Screening and Development: These cells are widely used for high-throughput screening of potential therapeutic compounds. By testing drugs on CFTR knockout HEK293T cells, scientists can identify molecules that may correct or compensate for the chloride transport defects that characterize cystic fibrosis, thereby facilitating the development of effective treatments.
3. Gene Therapy Studies: Researchers use CFTR knockout cell lines to evaluate the effectiveness of gene therapy strategies designed to correct the genetic defects of cystic fibrosis. These cells can evaluate a variety of vectors and delivery methods designed to restore CFTR function.
4. Mechanistic Studies of CFTR Function: Knockout cell lines are valuable for elucidating the functional role of CFTR in cellular processes. By comparing these cells to wild-type cells, scientists can gain insight into the physiological role of CFTR in various tissues and the consequences of its absence.
5. Biomarker Discovery: They aid in the identification of biomarkers of cystic fibrosis progression and treatment response. The unique cellular environment of CFTR knockout HEK293T cells can reveal novel biomarkers that are masked in the presence of functional CFTR.
For research use only. Not intended for any clinical use.
