Cat.No. : CSC-RR0493 Host Cell: HeLa
| Cat. No. | CSC-RR0493 |
| Description | HeLa-GFP/Luc is constructed by stable delivery of two vectors into HeLa, one vector containing Firefly luciferase reporter gene and puromycin resistance gene, the other vector containing EGFP reporter gene and neomycin resistance gene. HeLa cell line has been widely used in research. This cell line is an ideal positive control for use in both fluorescence and bioluminescence assays. |
| Gene | GFP/Luc |
| Host Cell | HeLa |
| Host Cell Species | Homo sapiens (Human) |
| Stability | Validated for at least 10 passages |
| Reporter Type | Fluorescent protein |
| 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 |
Lassa virus (LASV) can cause severe Lassa fever, for which there is currently no FDA-approved vaccine and limited treatment options. Previous studies have shown that the small matrix Z protein of LASV inhibits RIG-I-like receptors (RLRs), whereas the Z protein of the nonpathogenic Pichinde virus (PICV) does not, but its biological significance in infection remains unclear. In this study, the researchers developed a stable HeLa cell line (HeLa-iRIGN) that rapidly and robustly expresses the RIG-I N-terminal effector and produces type I interferons (IFN-Is) upon doxycycline (Dox) induction. They also generated recombinant tri-segmented PICVs (rP18tri-LZ and rP18tri-PZ), which encode LASV Z and PICV Z, respectively, along with a nonessential mScarlet fusion protein. After infection, rP18tri-LZ consistently exhibited higher viral gene expression and infection levels in HeLa-iRIGN cells compared to rP18tri-PZ, especially upon Dox induction. LASV Z significantly enhanced and prolonged viral gene expression in IFN-competent A549 cells. The study provides crucial insights into the biological role of LASV Z-mediated RIG-I inhibition and suggests that LASV Z may act as a virulence factor.
Figure 1. The researchers developed a HeLa-iRIGN that expresses RIG-I N-terminal effectors upon Dox induction, which allows for robust production of type I interferons. They employed an NDV-GFP-based assay to measure IFN levels by quantifying GFP expression in Vero cells treated with supernatants from Dox-induced HeLa-iRIGN cells. Additionally, they used reporter assays to evaluate the impact of LASV Z protein versus PICV Z protein on RIG-I inhibition, assessing the relative luciferase activity in response to different concentrations of these proteins. (Di D, et al., 2022)
Using Creative Biogene's GFP/Luc Reporter Cell Line - HeLa greatly simplifies similar experimental methods. The cell line has stable GFP and Luciferase reporter genes, which makes detecting IFN secretion and RIG-I inhibition easier and more efficient. Researchers can use integrated fluorescence and luminescence assays to swiftly and accurately measure protein function and interferon levels under a variety of circumstances, decreasing experimental time and complexity. With this cell line, you may easily acquire trustworthy data to help your research.
A: The GFP/Luc Reporter Cell Line - HeLa, with its ability to express both the green fluorescent protein (GFP) and the luciferase enzyme (Luc), allows researchers to simultaneously monitor two different biological processes. For instance, GFP expression can be used to track cellular localization or protein expression, while luciferase activity can report on the activity of specific promoters or signaling pathways. To ensure accurate and reliable data, it is essential to optimize the experimental conditions for both fluorescence and luminescence measurements. This includes selecting appropriate excitation/emission wavelengths for GFP, using luciferase substrates that do not interfere with GFP fluorescence, and normalizing the luciferase readings against GFP expression levels to account for variations in transfection efficiency or cell density.
A: The GFP/Luc Reporter Cell Line - HeLa is a valuable tool for assessing the efficiency of viral vectors in gene therapy research. By introducing a vector carrying the luciferase gene under the control of a specific promoter, researchers can measure transduction efficiency by the luminescence signal produced by the cells. The GFP expression serves as a visual confirmation of successful viral entry and gene expression. Critical parameters to control during these evaluations include the multiplicity of infection (MOI), the time post-transduction for analysis, and the background signal levels. Additionally, it is important to ensure that the viral vectors do not affect the cellular viability or the basal levels of GFP or luciferase expression.
A: To investigate the effects of small molecule inhibitors on cellular signaling pathways using the GFP/Luc Reporter Cell Line - HeLa, researchers can first establish a baseline of signaling activity by measuring the luciferase activity in untreated cells. Then, cells can be treated with the small molecules of interest, and the impact on signaling can be assessed by changes in luciferase activity. GFP expression can be used to monitor cell health and confluence throughout the experiment. For high-content screening, the assay can be scaled up to a 96- or 384-well format, allowing for the rapid evaluation of numerous compounds. Automated imaging systems and liquid handling robots can be employed to enhance throughput and accuracy.
A: The GFP/Luc Reporter Cell Line - HeLa can be used to study the regulation of gene expression by transcription factors by introducing a reporter construct containing the luciferase gene under the control of a promoter or enhancer element of interest. The activity of the transcription factor can then be measured by changes in luciferase activity. To validate these findings, molecular biology techniques such as chromatin immunoprecipitation (ChIP) assays, electrophoretic mobility shift assays (EMSAs), or RNA interference (RNAi) can be employed. These techniques can confirm the direct interaction of the transcription factor with the promoter/enhancer region and its effect on gene expression.
A: The GFP/Luc Reporter Cell Line - HeLa has significant potential applications in systems biology, where the goal is to understand the complex interactions and dynamics within cellular systems. By providing quantitative data on gene expression and signaling pathways, this cell line can inform the development of computational models that simulate cellular behavior in response to various stimuli. The dual-reporter system allows for the simultaneous analysis of multiple parameters, which is essential for building comprehensive models that can predict cellular responses under different conditions. The data generated can be integrated with other 'omics' data, such as proteomics and metabolomics, to create a more holistic representation of the cell's functional state and facilitate the identification of key regulatory nodes and potential therapeutic targets.
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This cell line incorporates both GFP and luciferase, allowing simultaneous fluorescence and luminescence reporting. This dual functionality enhances the utility of the GFP/Luc Reporter Cell Line - HeLa for complex biological assays.
The GFP/Luc Reporter Cell Line - HeLa can rapidly respond to external stimuli, enabling real-time observation of cellular reactions to treatments or environmental changes.
GFP and luciferase provide stable signal outputs in the GFP/Luc Reporter Cell Line - HeLa, which is crucial for longitudinal studies requiring consistent reporter activity over extended periods.
HeLa cells are known for their high transfection efficiency, making the GFP/Luc Reporter Cell Line - HeLa ideal for genetic manipulation experiments where introduction of additional genetic material is required.
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