Cat.No. : CSC-SC004308-RH Host Cell: HEK293
| Cat. No. | CSC-SC004308-RH |
| Description | This cell line is engineered to stably overexpress rat iodothyronine deiodinase 1 (Dio1, also known as 5DI, ITDI1, TXDI1). |
| Gene | Dio1 |
| Host Cell | HEK293 |
| Host Cell Species | Homo sapiens (Human) |
| Stability | Validated for at least 10 passages |
| Application | 1. Gene expression studies 2. Signaling pathway research 3. Drug screening and toxicology 4. Disease research |
| 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 |
The goal of the study was to examine the regulatory pathways that underlie the biological effects of thyroxine (T4). Because type 1 iodothyronine deiodinase (Dio1) is highly expressed in the liver, special attention was paid to this enzyme. They investigated the function of hepatocyte nuclear factor 4 alpha (HNF4α) in transcriptional regulation using the Dio1 cell line. Their research showed that the livers of HNF4α-null mice had significantly lower levels of Dio1 mRNA and enzyme activity, which led to higher serum concentrations of T4 and reverse T3 (rT3) but not T3. The transcriptional activation of the mouse Dio1 gene is significantly influenced by HNF4α, as confirmed by promoter analysis. Additionally, they showed that the HNF4α response element (HNF4α-RE) and the thyroid hormone-inducible Krüppel-like factor 9 (KLF9) effectively stimulate the mouse Dio1 promoter through two CACCC sequences. Furthermore, it was discovered that the synergistic activation of the Dio1 promoter required physical interactions between KLF9, HNF4α, and GATA4, indicating an indirect regulatory mechanism of Dio1 by thyroid hormone in mice. Together, these data imply that HNF4α, in cooperation with GATA4 and KLF9, transcriptionally modulates the mouse Dio1 gene to regulate thyroid hormone homeostasis.
Figure 1. The researchers subcloned the DNA fragment containing the 5'-FR of the mouse Dio1 gene into the pGL3 basic vector and transfected HEK293 and HepG2 cells to determine whether HNF4α can activate the Dio1 promoter. (Ohguchi H, et al., 2008)
A: Rat Dio1 Stable Cell Line-HEK293 has been tested for multiple passages and the expression of Dio1 remains stable. We conducted multiple passage experiments to verify the expression level of Dio1 through qPCR and Western Blot, and the results showed no significant changes.
A: We used qPCR and Western Blot to verify the expression of Dio1. The qPCR results showed that there was no significant difference in the expression levels of Dio1 mRNA in different passages. Western Blot results also showed that the expression level of Dio1 protein in each passage was consistent. Detailed experimental data and maps can be provided.
A: Rat Dio1 Stable Cell Line-HEK293 is suitable for thyroid hormone metabolism research, drug screening and related physiological function research. Functional validation included thyroid hormone deiodinase activity assays, which showed that Dio1 has the expected enzymatic activity in these processes. We also conducted relevant thyroid hormone level measurement experiments to confirm its functionality.
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The HEK293 Rat Dio1 stable cell line of this platform is really great, with consistent cell growth and stable gene expression, which is very suitable for my experimental needs.
The supplier's Rat Dio1 stable cell line is very good. The cell state is stable and the growth rate is fast, which makes my experiment progress smoothly.
The HEK293 cell line provided by the merchant has excellent performance, simple culture, strong cell viability, and high consistency of gene expression.
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