The ACE2 Knockout HCT 116 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population of HCT 116 cells with targeted disruption of the ACE2 gene. This polyclonal knockout pool provides a heterogeneous gene-disrupted background, enabling functional studies of angiotensin-converting enzyme 2 without the selection pressures inherent to clonal isolation. Loss of ACE2 protein and transcript can be routinely assessed by Western blotting and RT-qPCR, respectively, offering a flexible model for investigating ACE2-dependent processes in a colorectal carcinoma context.
The HCT 116 host cell line is a well-characterized human colorectal carcinoma epithelial model, notable for microsatellite instability-high (MSI-H) status and oncogenic mutations, including KRAS G13D, ??-catenin S45 deletion, and PIK3CA H1047R. These genetic alterations drive constitutive activation of key proliferative and survival pathways, making HCT 116 a robust system for dissecting intersecting signaling networks. The epithelial origin and colorectal background are particularly relevant for exploring ACE2 biology in gastrointestinal tissues, where ACE2 is normally expressed in the intestinal epithelium.
ACE2 is a carboxypeptidase that catalytically converts angiotensin II to angiotensin-(1-7), thereby negatively regulating the renin-angiotensin system. It also functions as the primary receptor for SARS-CoV and SARS-CoV-2 spike glycoproteins, mediating viral entry in cooperation with TMPRSS2 and furin. ACE2 interacts with the neutral amino acid transporter SLC6A19 (B0AT1) and integrin ITGA5, and its activity triggers downstream effectors such as the Mas receptor, PI3K/Akt pathway, ERK1/2 signaling, and eNOS activation. Acetylation, interferon signaling, hypoxia, and transcription factors including FoxA2, HNF1??, and GATA4 regulate ACE2 expression, while miRNAs like miR-421 and miR-200c post-transcriptionally modulate its levels.
Disruption of ACE2 in HCT 116 cells eliminates its enzymatic and receptor functions, creating a loss-of-function model to evaluate its contribution to tumor cell behavior. Given the mutational landscape of HCT 116 cells, the knockout may influence cellular proliferation, migration, and survival through altered angiotensin signaling crosstalk with oncogenic pathways. Additionally, this model allows interrogation of ACE2??s role in amino acid transport and gut homeostasis without the confounding effects of endogenous ACE2 activity, facilitating dissection of its non-catalytic functions in colorectal cancer biology.
This knockout tool is applicable to a variety of experimental scenarios, including SARS-CoV-2 pseudovirus entry assays for viral infection studies, angiotensin II/angiotensin-(1-7) ELISA for monitoring renin-angiotensin system modulation, and co-immunoprecipitation to map ACE2-SLC6A19 interactions. Further applications encompass drug screening for ACE2-targeted compounds, phospho-signaling analysis (Akt, ERK) to assess pathway activation, and migration/invasion assays to explore metastatic potential. Researchers can leverage these polyclonal cells for COVID-19 pathogenesis research and investigations into ACE2-dependent signaling in colorectal cancer. For additional details or to request a quote, please contact Ascent Research.
