The C1GALT1C1 Knockout HT29 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, in which the C1GALT1C1 gene has been functionally disrupted. This loss-of-function model enables the study of C1GALT1C1-dependent processes without defined clonal selection, providing a heterogeneous population that recapitulates diverse editing outcomes. The polyclonal format is particularly suited for pooled screening approaches and studies where clonal variation may influence phenotypic outcomes.
HT29 cells are a widely used human colorectal adenocarcinoma epithelial model with a hypertriploid karyotype and a homozygous R273H mutation in the TP53 tumor suppressor gene, which impairs p53-mediated DNA damage responses. Under appropriate culture conditions, HT29 cells can undergo enterocytic differentiation, making them valuable for investigating intestinal epithelial biology, barrier function, and the molecular pathology of colorectal cancer. The mutant p53 background also provides a relevant context for studying tumor-associated signaling and glycosylation alterations.
C1GALT1C1 serves as an essential molecular chaperone for C1GALT1 (core 1 synthase), and its functional disruption prevents proper folding and activity of C1GALT1, thereby blocking the conversion of Tn antigen to T-antigen, the initial step of mucin-type core 1 O-glycosylation. Consequently, loss of C1GALT1C1 leads to accumulation of truncated Tn antigen on glycoproteins such as MUC1, MUC2, integrin ??1, and CD44, and abrogates synthesis of extended O-glycan structures. The expression of C1GALT1C1 is subject to epigenetic silencing through promoter methylation and transcriptional regulation, and its activity directly modulates downstream O-glycosylation events that influence cell adhesion and immune signaling. Key pathway components include ppGalNAc-transferases, C1GALT1, T-synthase, and ST3GAL1, which collectively govern the biosynthesis of mucin-type O-glycans.
Within the HT29 colorectal cancer context, C1GALT1C1 disruption models the glycosylation defects observed in Tn syndrome and colorectal malignancies, where aberrant Tn antigen expression is associated with immune evasion, altered cell adhesion, and tumor progression. The HT29 background, with its mutant p53 and capacity for enterocytic differentiation, provides a unique platform to dissect the crosstalk between O-glycosylation and oncogenic signaling. This knockout model is particularly relevant for investigating how loss of core 1 O-glycosylation affects mucin barrier function, integrin-mediated adhesion, and interactions with the tumor microenvironment.
Researchers can use this polyclonal knockout model to study the role of core 1 O-glycosylation in colorectal cancer glycobiology, immune escape, and tumor-stroma interactions. Typical readouts include lectin blotting with PNA and VVA for Tn antigen, flow cytometry for Tn and T-antigen, Western blot for C1GALT1C1 and C1GALT1, and O-glycan mass spectrometry. Functional assays such as cell adhesion, migration, and invasion delineate the impact of altered glycosylation on metastatic behavior, while RNA-seq profiles glycosylation-related pathways. The product also supports mucin quantification and glycan-based drug targeting investigations. For further details, contact Ascent Research.
