Cat.No. : CSC-RR0534 Host Cell: A20
| Cat. No. | CSC-RR0534 |
| Description | This cell line is engineered to stably overexpress mCherry and luciferase reporter genes. A20 is a murine lymphoma cell line, which has been widely used in laboratory research. This cell line is a useful tool for both fluorescent and bioluminescent tracking of A20 cells. |
| Gene | mCherry/Luciferase |
| Host Cell | A20 |
| Host Cell Species | Mus musculus (Mouse) |
| Stability | Validated for at least 10 passages |
| Application | 1. Gene expression studies 2. Protein localization 3. Drug screening and toxicology 4. Live cell imaging |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| 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. |
| 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 |
The canonical NF-κB pathway is activated by LUBAC (linear ubiquitin chain assembly complex) through the linear polyubiquitination of NEMO (NF-κB essential modulator, also known as IKKγ) and RIP1. Researchers demonstrate that the Luciferase reporter cell line - A20 plays a crucial role in understanding the regulatory mechanisms of LUBAC-mediated NF-κB activation. Through binding linear polyubiquitin via the C-terminal seventh zinc finger (ZF7), A20 suppresses LUBAC-induced NF-κB activation. Crystal structure analysis reveals how A20 ZF7 recognizes linear diubiquitin, shedding light on genetic mutations linked to B cell lymphomas. Functional analysis highlights the importance of A20 ZF7 in recruiting A20 into TNFR signaling complexes, ultimately suppressing NF-κB activation.
Figure 1. The crucial role of A20 ZF7 in the suppression of LUBAC-mediated NF-κB activation was demonstrated. Various A20 mutants were examined, revealing that NF-κB activation induced by LUBAC was inhibited by A20 and CYLD, but not Cezanne. A20 ZFs, particularly ZF7, were found to significantly contribute to this suppression across different cellular contexts, including TNF-α-stimulated cells and MEFs overexpressing LUBAC, emphasizing the essential function of A20 ZF7 in NF-κB regulation. (Tokunaga F, et al., 2012)
A: A20 cells were chosen for establishing the mCherry/Luciferase Reporter Cell Line due to their relevance to lymphoma research and their well-characterized genetic background. Additionally, A20 cells are widely used in studies of B-cell biology and lymphoid malignancies, making them suitable for investigating the role of mCherry and luciferase in these cellular contexts.
A: Stable transfection techniques were employed to integrate the mCherry and luciferase reporter genes into the A20 cell genome, ensuring stable and consistent expression levels. The clonal selection was then performed to isolate cell lines with stable mCherry and luciferase expression. Regular monitoring of mCherry fluorescence and luciferase activity using appropriate assays ensured sustained expression levels over time.
A: Functional characterization of mCherry and luciferase expression in the A20 mCherry/Luciferase Reporter Cell Line involved studying their responsiveness to regulatory elements and their involvement in cellular processes relevant to lymphoma biology. This included investigating the activation of specific regulatory elements and monitoring changes in mCherry fluorescence and luciferase activity in response to signaling pathways dysregulated in lymphoma. Additionally, the roles of mCherry and luciferase in cellular processes such as proliferation, apoptosis, and drug response were assessed using molecular and cellular assays.
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The mCherry/Luciferase Reporter Cell Line in A20 cells combines mCherry fluorescence and luciferase bioluminescence, offering comprehensive insights into cancer biology.
The mCherry/Luciferase Reporter Cell Line simplifies experimental workflows, allowing for efficient data collection and analysis in cancer research. From studying tumor-immune interactions to evaluating treatment responses, this cell line serves as an invaluable resource for elucidating cancer biology and identifying novel therapeutic strategies.
With both reporters, this cell line enables detailed investigation of tumor growth, immune responses, and therapeutic efficacy, advancing our understanding of cancer progression. Its dual-reporter system provides multifaceted data, empowering deeper understanding of tumor dynamics and therapeutic targets.
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