The ACE2 Knockout PaTu 8988t Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal cell population with targeted disruption of the ACE2 gene in PaTu 8988t cells. This heterogeneous knockout pool enables loss-of-function studies while maintaining a genetic diversity that mimics native tumor heterogeneity. The cells are supplied as a ready-to-use model for investigating ACE2-dependent signaling and viral entry mechanisms without requiring clone isolation.
The parental PaTu 8988t line is an epithelial cell line derived from a liver metastasis of human pancreatic ductal adenocarcinoma (PDAC). It harbors KRAS G12V and TP53 mutations, central drivers of pancreatic cancer, and is widely used to study metastasis and tumorigenesis. These cells retain invasive and proliferative features characteristic of metastatic PDAC, providing a clinically relevant genetic background.
ACE2 encodes a transmembrane carboxypeptidase that serves as the receptor for SARS-CoV and SARS-CoV-2 spike proteins, with viral entry facilitated by cofactors TMPRSS2 and ADAM17. Within the RAS, ACE2 converts angiotensin II to angiotensin-(1-7), which binds Mas receptor to counterbalance AT1R-mediated signaling. ACE2 also forms a complex with SLC6A19 (B0AT1) for amino acid transport. Its expression is regulated by IFN-alpha, IFN-beta, androgen receptor, IL-1, TNF-alpha, and circadian components. Angiotensin-(1-7)?CMas receptor activation stimulates Akt and eNOS phosphorylation and anti-inflammatory cytokine release. Interacting partners such as integrins further modulate ACE2 output. Key RAS components??angiotensinogen, renin, angiotensin I, ACE, angiotensin II, AT1R, ACE2, angiotensin-(1-7), and Mas receptor??define the signaling cascade influenced by ACE2 knockout.
In PaTu 8988t cells, ACE2 knockout ablates the SARS-CoV-2 entry receptor and blocks conversion of angiotensin II to angiotensin-(1-7), leading to enhanced AT1R signaling and diminished Mas receptor activation. This shift may impact pancreatic cancer cell proliferation, invasion, and inflammatory responses, given the known roles of RAS in tumor progression. The oncogenic KRAS and TP53 mutant context allows interrogation of crosstalk between ACE2-regulated pathways and established tumorigenic drivers, offering insights into whether ACE2 loss exacerbates PDAC aggressiveness or alters the tumor microenvironment.
This knockout model enables diverse applications: studying SARS-CoV-2 infection in pancreatic cancer via pseudovirus entry assays and immunofluorescence; dissecting RAS signaling through western blotting, RT-qPCR, and RNA-seq; and testing ACE2-targeting drugs using angiotensin conversion assays. Proliferation (CCK-8) and invasion (transwell) assays assess functional impact, and cytokine ELISA profiles inflammatory mediators. The model also aids investigation of COVID-19-associated pancreatic complications and identification of therapeutic targets in PDAC. For further information, please contact Ascent Research.
