The HFE Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. This polyclonal pool contains a heterogeneous mixture of cells carrying targeted disruptions in the HFE gene, generating a loss-of-function model without clonal isolation. The product is designed for researchers studying iron homeostasis, intestinal epithelial biology, and related diseases.
The parental HT29 cell line, established from a human female colorectal adenocarcinoma, serves as a widely utilized model of intestinal epithelium. HT29 cells can be induced to differentiate into mucin-secreting goblet-like cells, enabling investigations of barrier function, nutrient absorption, and mucosal responses. Their epithelial origin and malignant phenotype make them suitable for colorectal cancer studies as well.
HFE encodes a homeostatic iron regulator that modulates cellular iron uptake by competing with transferrin for binding to transferrin receptor (TFRC). HFE also associates with transferrin receptor 2 (TFR2) and hemojuvelin (HJV) to enhance bone morphogenetic protein (BMP)-SMAD signaling. In this pathway, BMP6 binding to BMP receptors triggers phosphorylation of SMAD1/5/8, which transcriptionally upregulates hepcidin (HAMP) expression. Hepcidin, in turn, promotes internalization and degradation of ferroportin (SLC40A1), the sole iron exporter. Consequently, HFE disruption reduces hepcidin transcription, leading to increased ferroportin activity and altered cellular iron efflux. Downstream, iron storage proteins ferritin light chain (FTL) and ferritin heavy chain (FTH1) may be dysregulated, reflecting disrupted iron homeostasis.
In the HT29 intestinal epithelial context, HFE knockout provides a physiologically relevant platform to dissect iron handling mechanisms at the mucosal interface. Since intestinal epithelial cells are the primary site of dietary iron absorption, loss of HFE function in this model disrupts the normal regulatory loop linking cellular iron status to hepcidin production. This perturbation can be exploited to investigate how dysregulated iron homeostasis contributes to colorectal cancer progression, as iron accumulation promotes oxidative stress and proliferation. Moreover, the polyclonal nature preserves heterogeneous genetic backgrounds, allowing robust comparison of HFE-proficient and -deficient populations within the same experimental setting.
Typical applications include examining intestinal iron absorption kinetics using radiolabeled 59Fe uptake assays or calcein quenching fluorescence; quantifying hepcidin (HAMP) and ferroportin (SLC40A1) transcript levels by RT-qPCR; profiling iron-responsive proteins by western blotting for HFE, TFRC, and ferritin subunits; and assessing cell viability under iron overload conditions. These polyclonal knockout cells serve as a versatile tool for screening iron chelators or modulators and for RNA-seq-based transcriptome analyses to uncover novel iron-regulated genes. For further information or technical support regarding this product, please contact Ascent Research.
