Description

The TP53 Knockout Hep-G2 Cell Line is a CRISPR/Cas9-edited knockout cell line that provides a stable loss-of-function model of the TP53 tumor suppressor gene. Through CRISPR/Cas9-mediated gene disruption, this product enables researchers to study the consequences of p53 ablation in a human hepatocellular carcinoma background. The cell line serves as a robust tool for investigating p53-dependent signaling pathways and their roles in cell cycle regulation, apoptosis, and DNA damage responses.

The Hep-G2 cell line, derived from the liver hepatocellular carcinoma of a 15-year-old Caucasian male, is a widely used hepatic parenchymal model that recapitulates many aspects of liver metabolism and hepatocyte function. These cells retain key features of hepatic tissue, making them suitable for studying liver-specific gene functions and disease mechanisms. The TP53 knockout in this context effectively eliminates tumor suppressor activity, providing a clinically relevant model for hepatocellular carcinoma research.

TP53 encodes the transcription factor p53, a central tumor suppressor that integrates diverse cellular stress signals. Under DNA damage, upstream kinases ATM and ATR phosphorylate and activate p53, which is negatively regulated by MDM2-mediated degradation. Activated p53 transcriptionally upregulates key target genes: CDKN1A (p21) for cell cycle arrest, BAX and BBC3 (PUMA) for apoptosis, and GADD45A for DNA repair. p53 also interacts with cofactors p300/CBP and ASPP proteins to modulate transcriptional specificity. This pathway, involving ATM, ATR, CHK2, p53, MDM2, p21, BAX, and PUMA, constitutes a critical tumor-suppressive network frequently disrupted in cancer.

In Hep-G2 cells, TP53 knockout abrogates DNA damage checkpoint control, leading to unchecked proliferation and apoptosis resistance??phenotypes that mirror the loss of p53 function in hepatocellular carcinoma. This model enables dissection of p53-dependent tumor suppressor mechanisms in a hepatic context and evaluation of therapeutic strategies such as MDM2 inhibitors (e.g., Nutlin-3a), which require functional p53. It also provides a platform for studying synthetic lethal interactions and signaling rewiring in the absence of p53, contributing to liver cancer biology and drug discovery.

The TP53 Knockout Hep-G2 Cell Line is suitable for studying p53-dependent tumor suppression, screening p53 pathway modulators, and conducting liver cancer research. Representative assays include western blotting for p53 and targets, RT-qPCR for p21 and BAX, flow cytometry for apoptosis and cell cycle analysis, and drug sensitivity assays with compounds like Nutlin-3a. This line also supports xenograft studies to assess tumorigenicity and drug response in vivo. For protocols or inquiries, contact Ascent Research.