Human FGFR2 Knockout Cell Line-HEK293T
Cat.No. : CSC-RT0663
Host Cell: HEK293T Target Gene: FGFR2
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT0663 |
| Cell Line Information | A stable cell line with a homozygous knockout of human FGFR2 using CRISPR/Cas9. |
| Target Gene | FGFR2 |
| Host Cell | HEK293T |
| Shipping | 10^6 cells/tube |
| Storage | Liquid nitrogen |
| Species | Human |
| Gene ID | 2263 |
| 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
Case Study
Applications
Fibroblast growth factor receptor 2 (FGFR2) is a member of the fibroblast growth factor receptor family, which is known to play a key role in cell differentiation, growth, and regulation of various tissues. The FGFR2 gene is located on chromosome 10q26, and the protein it encodes is a receptor tyrosine kinase, which means that it has intrinsic enzymatic activity that transfers a phosphate group from ATP to a tyrosine residue in proteins. This role is critical for initiating downstream signaling pathway cascades that are important for cellular function.
Mutations and dysregulation of FGFR2 have been associated with a variety of human diseases. Germline mutations in FGFR2 can lead to a variety of developmental disorders, such as Crouzon syndrome, Apert syndrome, and Pfeiffer syndrome, all of which are types of craniosynostosis—disorders characterized by premature fusion of skull bones. These mutations often result in alterations in the receptor structure and function, leading to aberrant signaling. In addition to its role in developmental disorders, FGFR2 has been implicated in tumorigenesis. Somatic mutations, amplifications, and translocations of FGFR2 have been identified in a variety of cancers, including breast, endometrial, and gastric cancers. These alterations often result in constitutive activation of the receptor, promoting uncontrolled cell proliferation, survival, and migration. As a result, FGFR2 has become a target of interest for the development of novel cancer therapeutics, and several FGFR inhibitors are currently in clinical trials.
NF2 is a tumor suppressor gene that is frequently mutated in malignant pleural mesothelioma (MPM). Here, we found that cell growth, clonogenic activity, migration activity, and invasion activity of the NF2-knockout human mesothelial cell line MeT-5A (NF2-KO) were significantly increased compared with NF2-WT cell clones. Complementary DNA microarray analysis clearly revealed differences in the global gene expression profiles between NF2-WT and NF2-KO cell clones. Quantitative PCR analysis and Western blot analysis showed that upregulation of fibroblast growth factor receptor 2 (FGFR2) was concurrent with increased phosphorylation levels of JNK, c-Jun, and retinoblastoma (Rb) in NF2-KO cell clones. These increases were all abolished by exogenous NF2 expression in NF2-KO clones. Furthermore, disruption of FGFR2 in NF2-KO cell clones inhibited cell proliferation as well as the phosphorylation levels of JNK, c-Jun, and Rb. These findings suggest that NF2 deficiency may play a role in tumorigenesis of human mesothelial cells by mediating FGFR2 expression. FGFR2 will be a candidate molecule for the development of therapeutic and diagnostic strategies for NF2-deficient MPM.
MTT assays showed that the cell growth rate in NF2 and FGFR2 double knockout cell clones (NF2/FGFR2-DKO) was significantly reduced compared with NF2-KO clones (Figure 1A). In contrast, there was no significant change in the growth rate between FGFR2 knockout cells (FGFR2-KO) and parental cells (Figure 1B). In addition, disruption of FGFR2 in NF2-KO cells suppressed the NF2 knockout-induced migration and wound healing activities of NF2/FGFR2-DKO cells (Figures 1C,D). In addition, Western blot analysis showed that the phosphorylation levels of JNK and c-Jun were downregulated in NF2/FGFR2-DKO clones (Figure 1E). The researchers also found that the protein level of CDK2 and the phosphorylation level of Rb were reduced in NF2/FGFR2-DKO clones (Figure 1E). These results suggest that FGFR2 may play an important role in the proliferation of NF2-mutated mesothelioma cells.
Figure 1. Knockout of FGFR2 gene retards cell proliferation in the absence of NF2 gene. (Wahiduzzaman, Md, et al. 2019)
Knockout of the FGFR2 gene in HEK293T cells allows researchers to study the specific roles and mechanisms of FGFR2 in various cellular processes. Here are some key applications of the human FGFR2 knockout cell line - HEK293T:
Cancer Research: FGFR2 knockout cell lines are essential for studying the role of FGFR2 in cancer progression, as FGFR2 mutations and alterations have been implicated in various cancers. Researchers can assess how the loss of this receptor affects cellular behaviors such as proliferation, migration, and response to anticancer therapies.
Drug Development: These cell lines can be used to screen and validate new drugs targeting the FGFR2 pathway.
Signal Transduction Studies: FGFR2 is a key player in multiple signaling pathways. Knockout cell lines can be used to study how disruption of FGFR2 affects downstream signaling cascades, providing insights into cellular processes such as differentiation, survival, and metabolism.
Functional Genomics: Researchers use FGFR2 knockout cells to explore gene functions and interactions.
Tissue Engineering and Regenerative Medicine: FGFR2 is involved in tissue development and repair processes. Using knockout cell lines, scientists can develop better models to study tissue engineering, understand the molecular basis of regenerative medicine, and create new therapies for tissue injury and degeneration.
Disease Modeling: FGFR2 knockout cell lines are valuable models for studying congenital diseases and conditions associated with FGFR2 deficiency or dysregulation, such as craniosynostosis syndrome. It helps to elucidate the molecular basis of these conditions and test potential therapeutic strategies.
For research use only. Not intended for any clinical use.
