Human TNFRSF1A Knockout Cell Line-HeLa
Cat.No. : CSC-RT1286
Host Cell: HeLa Target Gene: TNFRSF1A
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT1286 |
| Cell Line Information | A stable cell line with a homozygous knockout of human TNFRSF1A using CRISPR/Cas9. |
| Target Gene | TNFRSF1A |
| Host Cell | HeLa |
| Shipping | 10^6 cells/tube |
| Storage | Liquid nitrogen |
| Species | Human |
| Gene ID | 7132 |
| 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 1:4-1:6. |
| 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
TNFRSF1A, also known as tumor necrosis factor receptor superfamily member 1A, is a gene encoding a TNF receptor that plays a key role in inflammation and apoptosis. TNFRSF1A is prominently expressed in many cell types, including immune cells, endothelial cells, and neuronal cells. The encoded protein is a key component in the TNF signaling pathway. TNFRSRF1A participates in mediating the effects of the proinflammatory cytokine tumor necrosis factor (TNF) by binding to TNF ligands. This interaction triggers a series of downstream signaling pathways that lead to a variety of cellular responses ranging from cell survival, apoptosis to inflammation.
Mutations in the TNFRSF1A gene are associated with TNF receptor-associated periodic syndrome (TRAPS), an autoinflammatory disease characterized by recurrent fever, abdominal pain, and rash. These mutations often result in abnormal folding and function of the receptor, leading to dysregulated inflammation and immune responses. In addition, TNFRSF1A has been implicated in other pathological conditions, including autoimmune diseases such as rheumatoid arthritis, multiple sclerosis, and inflammatory bowel disease. Therefore, understanding the mechanisms of TNFRSF1A is critical for developing targeted therapies for these diseases. For example, biologics that inhibit the interaction of TNF with TNFRSF1A have shown efficacy in treating rheumatoid arthritis and Crohn's disease.
Microtubule-targeting agents (MTAs) are one of the most widely used classes of chemotherapeutic drugs, and their mechanism of action has long been assumed to be blocking mitosis in rapidly dividing tumor cells. Contrary to this view, here researchers show that - in many cancer cell types - MTAs act by triggering membrane TNF (memTNF)-mediated cell-to-cell killing, which is very different from other non-MTA cell cycle blockers. Both memTNF and solTNF bind to TNF receptor 1 (TNFR1, also known as tumor necrosis factor receptor superfamily member 1A (TNFRSF1A)), thereby activating downstream cell death pathways. In cells that do not express RIP3 or have defective RIP3 signaling, TNFR1 activates the initiator protease caspase-8, which subsequently cleaves and activates the downstream executioner proteases caspase-3/7, leading to apoptosis. Here, researchers uncover a highly specific feature of the MTA family of drugs that distinguishes them from other mitotic blockers: both microtubule depolymerizers and stabilizers can directly induce TNF signaling-mediated tumor cell death, either by apoptosis in RIP3-deficient cancer cells or by necroptosis in RIP3-expressing cancer cells. Surprisingly, this MTA-induced cancer cell death was dependent on JNK/c-Jun-regulated accumulation of membrane TNF (memTNF) but not solTNF, through which it induces cancer cell killing by memTNF.
Most human cancer cells, such as HeLa cells, are resistant to necroptosis but sensitive to apoptosis due to the lack of expression of the necroptosis-essential gene RIP3. Here, the researchers tested the cell death response of human cancer cell lines lacking RIP3 to MTA. The study found that MTA also induced TNF-dependent apoptosis in HeLa cells. Blocking soluble TNF shedding by inhibiting TACE activity did not interfere with MTA-induced apoptosis; when TNFR1 was knocked out, cell death was abolished (Figure 1a-c). Ectopic expression of TNFR1 in TNFR1 (TNFRSF1A) knockout HeLa cells restored the apoptotic response to MTA (Figure 4d). The researchers next confirmed that MTA induced apoptosis in a variety of human cancer cell lines lacking RIP3 (Figure 1e-g). In fact, the mRNA levels of JUN and membrane-bound TNF in these human cancer cells were also upregulated by MTA treatment (Figure 1h,i).
Figure 1. MTAs induce memTNF-mediated apoptosis in RIP3-deficient human carcinoma cell lines. (Zhang, Jing, et al. 2020)
Cancer Research: The Human TNFRSF1A Knockout Cell Line-HeLa is essential for cancer research as it facilitates the study of the role of TNFRSF1A in tumorigenesis and cancer progression. By knocking out this receptor, researchers can observe changes in cell proliferation, apoptosis, and other cancer-related pathways.
Drug Development: This cell line is essential for screening potential therapeutics targeting the TNF signaling pathway. It enables the evaluation of drug efficacy and safety in the absence of TNFRSF1A, providing insight into alternative mechanisms of action and off-target effects.
Inflammatory Disease Models: This knockout cell line can be used to understand the role of TNFRSF1A in inflammatory processes.
Signal Transduction Studies: The TNFRSF1A Knockout Cell Line-HeLa serves as a model system to dissect the TNF signaling pathway. Researchers can study downstream signaling events, compensatory mechanisms, and crosstalk with other signaling pathways in detail, providing a comprehensive understanding of cellular communication processes.
Apoptosis Studies: By studying this knockout cell line, researchers can gain insight into the role of TNFRSF1A in programmed cell death.
For research use only. Not intended for any clinical use.
