InvivoGen offers a collection of NF-κB–SEAP reporter HEK293 cells to study the ALPK1-TIFA signaling axis.
Alpha protein Kinase 1 (ALPK1) and TRAF interacting forkhead-associated protein A (TIFA) form an important cytoplasmic surveillance pathway of pathogenic Gram-negative bacteria such as Shigella flexneri and Helicobacter pylori [1,2].
ALPK1 is a critical cytoplasmic alpha kinase responsible for the recognition and binding of bacterial sugar metabolites. ALPK1 recognizes and binds directly to the metabolic intermediates in lipopolysaccharide (LPS) biosynthesis, the potent ADP-β-d-manno-heptose (ADP-Heptose) and the enzymatically converted d-glycero-β-d-manno-heptose-1,7-bisphosphate (HBP) [3]. ADP-Heptose and HBP can be delivered to the host cell cytoplasm in a number of different ways including type III and IV secretion systems (e.g. Yersinia pseudotuberculosis and H. pylori, respectively), as well as by endocytosed bacteria [2-3]. Notably, ADP-Heptose can freely penetrate the cell membrane, whereas HBP requires the use of pore-forming agents [3, 4.
Activated ALPK1 phosphorylates TRAF interacting forkhead-associated protein A (TIFA), inducing its oligomerization and the formation of 'TIFAsomes', which in turn recruits and activates TRAF6. Ultimately, the ALPK1-TIFA signaling axis results in the initiation of an NF-κB dependent pro-inflammatory response [1-4].
Targeting this pathway will be important in the development of novel treatments for modulating inflammation caused by bacterial infection as well as diseases associated with its dysregulation such as gastric cancer.
To foster research on this pathway, InvivoGen provides cellular tools that have been generated from the parental HEK-Blue™ Null1-v Cells, a human embryonic kidney (HEK)-293 derived cell line, through the stable knockout (KO) of the ALPK1 or TIFA gene. These cell lines express a secreted embryonic alkaline phosphatase (SEAP) under the control of an NF-κB-inducible promoter. Incubation of HEK-Blue™ Null1-v cells with ADP-Heptose induces the activation of the NF-kB pathway and the production of SEAP, whereas HEK-Blue™ KO-ALPK1 cells and HEK-Blue™ KO-TIFA cells are unresponsive to ADP-Heptose stimulation.
The collection comprises
- HEK-Blue™ KO-ALPK1 Cells – Knockout (KO) of the ALPK1 gene
- HEK-Blue™ KO-TIFA Cells – Knockout (KO) of the TIFA gene
- HEK-Blue™ Null1-v – Parental cell line
Key Features
- Verified KO of the ALPK1 or TIFA gene (DNA sequencing, PCR, and functional assays)
- Readily assessable NF-κB-dependent SEAP reporter activity
- Each lot is functionally tested and guaranteed mycoplasma-free
InvivoGen also offers ADP-L-Heptose, an AhR ligand.
