ACE2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human colorectal adenocarcinoma HT29 cell line. This loss-of-function model disrupts the ACE2 gene, enabling robust investigation of ACE2-dependent cellular processes without generating a monoclonal line. The polyclonal nature preserves genetic heterogeneity, reflecting a more physiologically relevant knockout context.
The HT29 cell line is a well-characterized human colorectal adenocarcinoma model originally established from a primary tumor of a male donor. These cells exhibit an intestinal epithelial phenotype and are extensively employed in colorectal cancer research, studies of intestinal barrier function, and inflammatory bowel disease models. Their adherent growth and responsiveness to extracellular stimuli make them suitable for a wide range of biochemical and cell-based assays.
ACE2 encodes a carboxypeptidase that cleaves angiotensin II to angiotensin-(1-7), thereby counterbalancing the pro-inflammatory and vasoconstrictive angiotensin II/AT1 receptor axis through activation of the Mas receptor. Additionally, ACE2 acts as the primary receptor for SARS-CoV-2 viral entry via binding the spike protein, facilitated by TMPRSS2, and functions as a chaperone for the neutral amino acid transporter B(0)AT1 (SLC6A19). ACE2 expression and activity are regulated by interferon signaling, HNF4??, GATA4, angiotensin II, TNF-??, IL-6, and bile acids, while downstream mediators include nitric oxide, anti-inflammatory cytokines, and AT2 receptor signaling. ACE2 interacts with ADAM17, calmodulin, and integrins, linking its function to diverse pathways such as mTOR and amino acid transport.
In the context of HT29 cells, ACE2 modulates local angiotensin signaling that can influence colorectal tumor cell proliferation, migration, and inflammatory responses. Given the intestinal origin, this knockout model is particularly relevant for dissecting ACE2’s role in epithelial barrier integrity, gut amino acid absorption, and host-pathogen interactions at the mucosal surface. Loss of ACE2 may shift the balance between pro- and anti-inflammatory signals, impacting tumor microenvironment dynamics and intestinal homeostasis.
This polyclonal knockout cell population is a versatile tool for multiple research applications. It is ideal for studying SARS-CoV-2 infection mechanisms through pseudovirus entry assays and for evaluating ACE2-targeted therapeutics. The cells support detailed analysis of ACE2-dependent signaling in colorectal cancer using western blot, RT-qPCR, immunofluorescence, and angiotensin II conversion assays. Further uses include drug screening for renin-angiotensin system modulators, co-immunoprecipitation studies with spike protein or endogenous ACE2 partners, and functional assays such as migration/invasion and flow cytometry for surface ACE2 expression. RNA-seq can be performed to assess transcriptome-wide effects of ACE2 disruption. For more information, please contact Ascent Research.
