Cat.No. : CSC-RR0573 Host Cell: HEK293
| Cat. No. | CSC-RR0573 |
| Description | The overexpression cassette containing GFP reporter gene driven by a tetracycline inducible promoter (Tet-On system) was stably integrated into the HEK293 genome. GFP expression is rapidly induced in the presence of tetracycline (Tc) or tetracycline-derivatives like doxycycline (dox). |
| Gene | Tet-On GFP |
| Host Cell | HEK293 |
| Host Cell Species | Homo sapiens (Human) |
| 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. |
| Media Type | Cells were cultured in DMEM supplemented with 10% fetal bovine serum. |
| Freeze Medium | Complete medium supplemented with 10% (v/v) DMSO |
| 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. |
| 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. |
| 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 |
In the realm of cellular biology, the intricate interactions between intracellular proteins play pivotal roles in both normal physiology and various diseases. Traditional methods aimed at modifying these pathological protein-protein interactions often face significant challenges. Researchers are now turning to innovative solutions like bioPROTACs—engineered proteins that facilitate targeted degradation of therapeutic targets by fusing a target-binding domain with an E3 ligase. This study highlights the adaptability of bioPROTACs in targeting proteins such as proliferating cell nuclear antigen (PCNA), showcasing their potential for therapeutic applications and biological research.
Figure 1. The researchers employed flow cytometric analysis using H2B-GFP/HEK293 Tet-On 3G cells to evaluate the efficacy of various GFP binders. By inducing mCherry expression with doxycycline, they aimed to assess the flexibility of bioPROTAC design in facilitating targeted protein degradation. (Lim S, et al., 2020)
Creative Biogene's Tet-On GFP Stable Cell Line - HEK293 offers a versatile platform for similar targeted degradation studies. With its ability to stably express GFP-tagged proteins, this cell line enables researchers to easily visualize and analyze protein interactions and degradation processes.
A: The Tet-On GFP Stable Cell Line - HEK293 allows researchers to control the expression of GFP through the addition or removal of tetracycline or its derivatives, such as doxycycline. This system enables the study of protein localization and trafficking by providing a means to temporally regulate GFP fusion proteins. Researchers can observe the effects of GFP expression on cellular processes without the need for labor-intensive and time-consuming protein tagging for each individual study, thus streamlining the research process and allowing for more dynamic analysis of protein behavior within living cells.
A: The Tet-On GFP Stable Cell Line - HEK293 can be engineered to express genes of interest under the control of the inducible promoter. By inducing GFP expression, researchers can monitor cell cycle progression and proliferation rates in real-time. The system can be further adapted to express proteins that are known to affect cell cycle regulation, allowing for a detailed analysis of how these proteins influence cell cycle dynamics. This approach can lead to a better understanding of the molecular mechanisms that govern cell growth and division.
A: The Tet-On GFP Stable Cell Line - HEK293 serves as an excellent model for evaluating the safety and efficacy of inducible gene expression systems for potential gene therapy applications. By using this cell line, researchers can test the responsiveness of the inducible promoter system to various inducers and assess the stability and long-term expression of the transgene. Additionally, the system can be used to evaluate potential off-target effects and the impact of gene expression on cellular function, which are critical considerations for the development of safe and effective gene therapies.
A: The Tet-On GFP Stable Cell Line - HEK293 can be utilized to investigate how different cellular conditions, such as stress, differentiation, or exposure to various chemical agents, affect gene regulation and epigenetic modifications. By inducing GFP expression and monitoring changes in chromatin structure and histone modifications, researchers can gain insights into the epigenetic mechanisms that control gene expression. This knowledge can contribute to the understanding of how environmental factors influence gene regulation and cellular behavior.
A: The Tet-On GFP Stable Cell Line - HEK293 can be used to fine-tune inducible expression systems for biotechnological and industrial purposes. By testing the responsiveness of the cell line to different inducers and varying concentrations, researchers can optimize the expression system for maximum efficiency and minimal toxicity. This cell line can also be used to screen for novel inducers or to engineer cells with improved inducible expression capabilities, which can be beneficial for the production of recombinant proteins or the development of cell-based biosensors.
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The Tet-On GFP Stable Cell Line - HEK293 offers inducible GFP expression, allowing us to control when GFP is expressed by adding or removing doxycycline. This control is crucial for temporal studies where the timing of gene expression is critical to the experimental outcomes.
With GFP as a reporter, the Tet-On GFP Stable Cell Line - HEK293 enables visual tracking of gene expression. This visualization is invaluable for confirming the presence and location of gene expression in real time, facilitating immediate observational studies.
HEK293 cells are known for their high transfection efficiency, making the Tet-On GFP Stable Cell Line - HEK293 ideal for introducing additional genetic materials. This characteristic is beneficial for experiments requiring the co-expression of other genes alongside GFP.
The Tet-On GFP Stable Cell Line - HEK293 benefits from the robust growth and easy maintenance characteristics of HEK293 cells, which supports a wide range of experimental conditions and reduces the complexity of cell culture management.
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