Cat.No. : CSC-RR0082 Host Cell: HEK293
| Cat. No. | CSC-RR0082 |
| Introduction | The cell line demonstrates strong GFP fluorescent signal in normal culture condition as the constitutive CMV promoter drives the GFP high expression to the stop codon. The downstream RFP ORF was not expressed because of the stop codon after the GFP ORF. Once the CRE protein was present in nuclear, the CRE excises / deletes the DNA fragment between two loxP sites, which removes the stop codon. As a result, the RFP ORF is then expressed under the CMV promoter, and the cell line switches to RFP fluorescent. The RFP signal can be easily monitored via fluorescent cell sorting (for the ratio between GFP and RFP cells), or visualized under microscope, or measured the fluorescent intensity by a meter or reader with RFP filter set. |
| Gene | GFP/RFP |
| Host Cell | HEK293 |
| Host Cell Species | Homo sapiens (Human) |
| Morphology | Epithelial |
| 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. |
| 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 |
RNA serves as a central information carrier within cells, providing a robust interface for both observing and directing cellular behaviors. Sequencing RNA allows for precise readout and understanding of cell states. The loxP/GFP/RFP Color-Switch Reporter Stable Cell Line-HEK293 was created to enable dynamic RNA export from live cells. This technique makes use of viral mechanism-inspired genetically encoded RNA exporters. It permits target RNA molecules to be specifically packaged and secreted from mammalian cells in protective nanoparticles. It enables non-destructive monitoring of cell dynamics at clonal resolution through the exportation and sequencing of RNA barcodes. By facilitating cell-to-cell RNA transmission, fusogen incorporation promotes hybrid cell and gene therapies by facilitating the delivery, expression, and functional activity of exported mRNA in destination cells. Known as COURIER (Controlled Output and Uptake of RNA for Interrogation, Expression, and Regulation), this technology facilitates the delivery of tailored RNA programs as well as real-time cell monitoring.
Figure 1. HEK293 Color-Switch loxP/GFP/RFP cells (referred to as Cre reporter cells) from Creative Biogene were seeded into 24-well plates at a density of 50,000 cells per well. The researchers demonstrate the utility of the loxP/GFP/RFP Color-Switch Reporter Stable Cell Line-HEK293 in visualizing RNA delivery dynamics. By co-culturing sender cells expressing a Cre mRNA cargo and optimized delivery system with receiver cells, they observe efficient RNA transfer. This system enhances delivery across the receiver cell population, facilitating robust cellular response monitoring. (Horns F, et al., 2023)
A: Our loxP/GFP/RFP CRE Reporter cell line has been rigorously screened and verified to ensure stable expression of GFP and RFP. We verified the expression levels of GFP and RFP through fluorescence microscopy observation and flow cytometry, and performed repeated tests at different time points and in different batches. Cells express GFP and RFP under appropriate culture conditions and maintain stable expression levels.
A: Our loxP/GFP/RFP CRE Reporter cell line has been rigorously screened and verified to ensure activation of the CRE gene. We verified the activation level of the CRE gene through methods such as PCR and fluorescence microscopy observation, and conducted repeated tests at different time points and in different batches. Cells maintain the activation state of the CRE gene under appropriate culture conditions and exhibit stable activation levels.
A: Our loxP/GFP/RFP CRE Reporter cell line has undergone a rigorous selection and optimization process. We use specific screening criteria, including fluorescence microscopy and gene expression levels, to ensure that the stability and expression levels of the cell lines meet our quality standards. We also performed long-term culture and expansion of the cell line to verify its stability and reproducibility under different conditions.
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I am very pleased with the use of this loxP/GFP/RFP (Puromycin) Color-Switch cell line. In my experiments, these cells showed impressive stability. They exhibit consistent growth characteristics and stable gene expression levels both in culture and in experimental operations. By tracking and measuring the fluorescence of GFP and RFP, I successfully observed the occurrence of genetic recombination events, which provided an important visual reference for my research. These cells also possess durable anemiamycin resistance, making it easier to select and maintain stable transgenic cells in experiments. The use of this cell line makes my experiments smoother and more efficient.
HEK293 cells are ideal experimental materials because they are easy to culture and transfect, and they grow well under conventional tissue culture conditions. This allows me to be flexible in my experiments and easily introduce exogenous genetic material for functional studies. Through this cell line, I successfully explored the gene recombination process mediated by Cre recombinase, which provided valuable data support for my research.
I would like to emphasize the great help that loxP/GFP/RFP (Puromycin) Color-Switch has brought to my research. The stability and operability of this cell line are excellent. In my experiments, I observed that these cell lines not only maintained stable growth during culture, but also showed excellent consistency at the gene expression level. By tracking the fluorescence signals of GFP and RFP, I successfully studied genetic recombination events, which provided intuitive data support for my experiments. Furthermore, these cells are also anemicin resistant, allowing me to easily select and maintain stable transgenic cells.
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