ACE2 Knockout HuH-7 Cell Line

The ACE2 Knockout HuH-7 Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the HuH-7 human hepatocellular carcinoma cell line. It is engineered for targeted disruption of the ACE2 gene, resulting in loss of functional ACE2 protein. This model provides a stable, isogenic platform for interrogating ACE2-dependent signaling, viral entry, and liver pathobiology in a disease-relevant context.

The parental HuH-7 cell line is a well-differentiated, epithelial hepatocellular carcinoma derived from a 57-year-old Japanese male in 1982. It harbors a p53 mutation and is extensively characterized for liver cancer research, hepatitis virus replication (including HCV and HBV), and drug metabolism assays. Its human hepatic origin and retention of liver-specific functions such as lipoprotein synthesis and xenobiotic metabolism render it a relevant host for studying ACE2-mediated liver fibrosis, inflammation, and viral interactions.

ACE2 is a type I transmembrane metalloprotease that functions as a critical modulator of the renin-angiotensin system (RAS). Its carboxypeptidase activity cleaves angiotensin II to generate angiotensin (1-7), a heptapeptide that signals through the Mas receptor to activate the Akt and ERK pathways, stimulate nitric oxide production, and oppose AT1 receptor-driven vasoconstriction, inflammation, and fibrogenesis. ACE2 expression is positively regulated by inflammatory cytokines: interferon-gamma and interferon-alpha induce transcription via STAT1, while IL-1?? and TNF-?? act through HNF1A and NF-??B-dependent mechanisms. Additionally, ACE2 serves as the obligate entry receptor for SARS-CoV-2, binding the viral spike glycoprotein and forming a complex with the amino acid transporter B0AT1 (SLC6A19) and integrin ??5??1.

In the HuH-7 knockout context, disruption of ACE2 eliminates both its enzymatic and receptor functions. Loss of angiotensin (1-7) production is expected to elevate local angiotensin II levels, shifting the RAS balance toward pro-inflammatory and pro-fibrotic AT1 receptor signaling??a key driver in hepatic stellate cell activation and liver fibrosis progression. Concurrent absence of the viral receptor confers resistance to SARS-CoV-2 entry, making these cells a valuable tool for studying host-cell responses to viral challenge without confounding infection, and for dissecting COVID-19-associated liver injury mechanisms.

The ACE2 Knockout HuH-7 Cell Line is well-suited for SARS-CoV-2 pseudovirus entry assays, angiotensin (1-7) quantification by ELISA, and expression analysis of ACE2 via western blotting and RT-qPCR. Researchers can also evaluate ACE2 enzymatic activity, monitor surface ACE2 by flow cytometry or immunofluorescence, and measure cytokine responses using multiplex profiling. Additionally, cell migration, invasion, and fibrosis marker expression can be assessed to study ACE2’s role in liver pathology. The line is amenable to drug screening for ACE2 inhibitors or antiviral compounds. For further product details or technical inquiries, contact Ascent Research.