The ACYP2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HT29 human colorectal adenocarcinoma cells, designed to disrupt ACYP2 gene function. This heterogeneous pool carries diverse loss-of-function mutations, enabling population-level studies of acylphosphatase-2 deficiency without clonal bias. The polyclonal format is suitable for pooled functional analyses including butyrate-induced differentiation and ion transport assays.
The HT29 cell line originates from a primary colorectal adenocarcinoma and contains a KRAS G13D mutation along with mutant APC and TP53. These genetic alterations make HT29 a standard model for colorectal cancer progression and epithelial differentiation. HT29 cells can be induced to differentiate along the enterocytic lineage by butyrate or 1,25-dihydroxyvitamin D3, a process that relies on functional acylphosphatase-2.
Acylphosphatase-2, encoded by ACYP2, hydrolyzes acyl phosphates and modulates Na+/K+-ATPase activity and cellular differentiation. In HT29, ACYP2 is regulated upstream by butyrate, CDX2, and 1,25-dihydroxyvitamin D3, and it acts downstream to promote alkaline phosphatase expression. ACYP2 physically interacts with Na+/K+-ATPase and carbamoyl phosphate, linking phosphate metabolism to ion homeostasis and enterocyte maturation. Knockout of ACYP2 eliminates this enzymatic activity, impairing ion pump regulation and altering proliferation-differentiation balance.
In the HT29 context, ACYP2 knockout provides a model to study its potential tumor suppressor role. Loss of ACYP2 is expected to attenuate butyrate-induced differentiation, evidenced by reduced alkaline phosphatase and villin, while possibly enhancing malignant features driven by KRAS and TP53 mutations. The polyclonal pool reflects tumor heterogeneity, adding relevance for differentiation-based therapy studies.
Applications include colorectal cancer research, intestinal differentiation studies, ion transport regulation, and drug response assays. Typical methods: western blotting, RT-qPCR, alkaline phosphatase activity, ion flux measurements, villin immunofluorescence, and proliferation assays under butyrate stimulation. For further details, contact Ascent Research.
