TRIB3 Knockout Hep-G2 Cell Line

TRIB3 Knockout Hep-G2 Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the Hep-G2 human hepatocellular carcinoma cell line, providing a targeted gene disruption model for the stress-responsive pseudokinase TRIB3. This product enables researchers to study the loss-of-function consequences of TRIB3 in a well-characterized hepatic cellular context, without the need for transient knockdown approaches or pharmacological inhibition.

The Hep-G2 cell line originates from the liver hepatocellular carcinoma of a 15-year-old male and serves as a widely used hepatic epithelial model. These cells retain many characteristics of primary human hepatocytes, including expression of liver-specific enzymes and responsiveness to metabolic and toxic stimuli, making them valuable for investigations into liver metabolism, drug-induced hepatotoxicity, and viral hepatitis pathogenesis.

TRIB3 is a pseudokinase upregulated by endoplasmic reticulum (ER) stress and nutrient deprivation through the PERK-eIF2??-ATF4-CHOP signaling axis. As a scaffold protein, TRIB3 directly binds to and inhibits Akt1, Akt2, and Akt3, thereby impairing downstream insulin signaling and promoting gluconeogenesis via dephosphorylation of FOXO1 and GSK3??. TRIB3 also interacts with ATF4 and CHOP to enhance ER stress-induced apoptosis, and its expression is stimulated by inflammatory cytokines such as TNF?? and IL-6, linking cellular stress responses to insulin resistance and metabolic dysfunction.

In the Hep-G2 hepatocellular carcinoma context, knockout of TRIB3 disrupts the stress-induced inhibition of Akt, potentially restoring insulin signaling and attenuating ER stress-driven apoptosis. This makes the TRIB3 Knockout Hep-G2 Cell Line a powerful tool for dissecting the molecular crosstalk between ER stress, insulin resistance, and hepatic lipid metabolism. Researchers can utilize this model to examine how TRIB3 loss modulates gluconeogenic gene expression, lipid droplet accumulation, and cell survival under metabolic or pharmacologic stress.

This knockout cell line is suitable for a range of applications, including but not limited to exploring hepatic insulin resistance pathways by measuring Akt phosphorylation and glucose output; investigating ER stress responses through tunicamycin treatment followed by analysis of CHOP and ATF4 levels; examining hepatocellular carcinoma cell survival and apoptosis via Annexin V/PI staining; and assessing lipid metabolism by oil red O staining. Additionally, co-immunoprecipitation experiments can be performed to confirm altered protein interactions in the absence of TRIB3. For detailed product information, protocols, or technical support, please contact Ascent Research.