The ECE1 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population engineered to disrupt the ECE1 gene in the human hepatocellular carcinoma cell line Huh-7. This pooled loss-of-function model enables investigation of endothelin-converting enzyme 1 (ECE1) biology without clonal selection biases. The polyclonal format maintains genetic diversity, making it a robust tool for population-level analyses of ECE1 signaling pathways. This product serves as a reliable resource for functional studies and drug screening in endothelin biology.
Huh-7 cells originate from a well-differentiated hepatocellular carcinoma of a 57-year-old Japanese male. They harbor a mutant p53 (Y220C) and are permissive for hepatitis C virus (HCV) replication, making them an essential model for hepatocarcinogenesis, virus?Chost interaction studies, and drug metabolism research. As a liver parenchymal cell line, Huh-7 provides a clinically relevant hepatocellular context for functional genomics, drug testing, and signal transduction studies.
ECE1 encodes a zinc metalloprotease that converts big endothelins (big ET-1, -2, -3) to active endothelin-1 (ET-1). ET-1 then binds the G protein-coupled receptors EDNRA and EDNRB, triggering downstream signaling via GNAQ, PLCB, calcium mobilization, and the MAPK/ERK cascade (MAPK1/MAPK3). Expression is induced by pro-inflammatory and fibrotic stimuli such as TNF??, IL-1??, TGF-??, and hypoxia, and ET-1 participates in autoregulatory feedback. Collectively, this signaling network controls vasoconstriction, cell proliferation, and matrix deposition??processes dysregulated in liver fibrosis and portal hypertension.
Disruption of ECE1 in Huh-7 cells abrogates endogenous ET-1 production, permitting direct dissection of the ET-1/EDNR signaling module in hepatocellular carcinoma. This knockout model is particularly valuable for investigating hepatic stellate cell activation and the fibrogenic gene expression programs central to liver fibrosis and portal hypertension. Furthermore, the HCV-permissive background of Huh-7 allows exploration of potential crosstalk between hepatitis C virus infection and the endothelin pathway, which may reveal viral modulatory effects on ECE1 activity or downstream fibrotic mediators.
This polyclonal ECE1 knockout pool supports diverse functional assays. ET-1 secretion can be quantified by ELISA, while EdU incorporation and wound healing assays assess proliferation and migration. qRT-PCR and western blotting detect changes in fibrotic markers such as COL1A1 and ??-SMA. MAPK/ERK activation is measured by phospho-kinase assays, and calcium flux assays monitor EDNR-mediated signaling. Additional tools include co-immunoprecipitation for substrate interactions, RNA-seq for transcriptomic profiling, and flow cytometry for apoptosis or cell cycle analysis. This versatile resource enables comprehensive investigation of endothelin biology in liver cancer, fibrosis, and HCV pathogenesis. For further information or to discuss custom knockout services, please contact Ascent Research.
