Cat.No. : CSC-RT0451
Host Cell: HEK293T Target Gene: TYK2
Size: >1x10^6 cells/vial Validation: Sequencing
| Cat. No. | CSC-RT0451 |
| Cell Line Information | This cell line is a stable cell line with a homozygous knockout of human TYK2 using CRISPR/Cas9. |
| Target Gene | TYK2 |
| Gene ID | 7297 |
| Genotype | TYK2 (-/-) |
| Host Cell | HEK293T |
| Cell Type | Epithelial |
| Size | >1x10^6 cells/vial |
| Sequencing Result | Homozygous: 1 bp insertion in exon2 |
| Species | Homo sapiens (Human) |
| 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. |
| Media Type | Cells were cultured in DMEM supplemented with 10% fetal bovine serum. |
| 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. |
| Freeze Medium | Complete medium supplemented with 10% (v/v) DMSO |
| 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 |
Deciphering the complex dynamic events that control type I interferon (IFN) signaling is critical to uncovering key regulatory mechanisms in host antiviral defense. Here, researchers used TurboID-based proximity tagging combined with affinity purification mass spectrometry to comprehensively map the proximal human proteome of all seven canonical type I IFN signaling cascade members under basal and IFN-stimulated conditions. This revealed 103 high-confidence protein networks tightly associated with core members IFNAR1, IFNAR2, JAK1, TYK2, STAT1, STAT2, and IRF9, and validated several known constitutive protein assemblies, while also revealing novel stimulus-dependent and stimulus-independent associations between key signaling molecules. Mechanistically, PJA2 interacts with TYK2 and JAK1, promotes their non-degradative ubiquitination, and limits activating phosphorylation of TYK2, thereby inhibiting downstream STAT signaling. These high-resolution proximal protein maps provide global insights into the type I IFN signaling network and serve as a valuable resource for future exploration of its functional complexity.
TYK2 has been previously shown to stabilize IFNAR1 on the surface of unstimulated cells by preventing its internalization and turnover through endocytosis. In this study, the researchers used a HEK293T-based TYK2 KO cell line to confirm that the lack of TYK2 reduced the total level of IFNAR1 (Figure 1c). In addition, re-expression of TYK2 partially restored IFNAR1 protein levels, but further expression of PJA2-WT could not reverse this phenomenon (Figure 1c, d), indicating that PJA2-promoted TYK2 ubiquitination does not interfere with TYK2's ability to stabilize IFNAR1 levels.
Figure 1. c NE (non-edited) HEK293T cell clones or TYK2 KO (knockout) cell clones were transfected with the indicated plasmids, and total cell lysates were analyzed by SDS-PAGE and immunoblotting. d Quantification of IFNAR1 protein expression normalized to β-actin protein expression and made relative to one NE control clone in replicates of panel (c). (Schiefer, Samira, and Benjamin G. Hale. 2024)
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