The AFTPH Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human colorectal adenocarcinoma HT29 cell line, engineered to disrupt the AFTPH gene. This pool provides a genetically heterogeneous loss-of-function model for investigating clathrin-mediated endocytosis and associated intracellular trafficking pathways in a colorectal cancer background. The polyclonal nature offers a robust population-level knockout without clonal selection, enabling studies of AFTPH-dependent processes with reduced clonal artifacts.
HT29 is a well-characterized epithelial cell line originally isolated from a colorectal adenocarcinoma of a 44-year-old female. These cells are widely employed as a model system for intestinal epithelial cell biology, differentiation, and colorectal cancer research. They retain key features of intestinal enterocytes, including the ability to polarize and form tight junctions, making them suitable for examining endocytic trafficking in the context of polarized epithelial cells and intestinal barrier function.
AFTPH encodes a clathrin adaptor protein that plays a critical role in cargo sorting and receptor-mediated endocytosis. It functions by interacting with the AP2 complex (specifically AP2B1) and clathrin heavy chain to promote the recruitment of clathrin coats to the plasma membrane and facilitate the internalization of transmembrane receptors. AFTPH operates downstream of receptor tyrosine kinase signaling and clathrin coat assembly, and its activity influences downstream targets such as the MAPK/ERK signaling cascade. Knockout of AFTPH disrupts the endocytic machinery, impairing the internalization of diverse cargo including transferrin and epidermal growth factor receptor (EGFR), thereby altering receptor recycling, signal transduction, and cellular responses.
In the context of HT29 colorectal adenocarcinoma cells, disruption of AFTPH-mediated endocytosis provides a powerful tool for dissecting how altered membrane trafficking contributes to cancer phenotypes. Colorectal cancer cells often exhibit dysregulated receptor signaling and endocytic pathways, and this knockout model enables systematic interrogation of the role of clathrin adaptor proteins in controlling receptor surface levels, proliferative signaling, and drug delivery. The availability of this polyclonal population facilitates the study of endocytosis-related disorders and the identification of trafficking-dependent vulnerabilities in colorectal cancer cells.
This product is suitable for a wide range of experimental applications, including the mechanistic investigation of clathrin-mediated endocytosis in colorectal cancer, assessment of intracellular trafficking and receptor recycling, and studies of drug delivery vehicle uptake. Representative assays include transferrin and EGFR internalization assays, immunofluorescence staining for clathrin and AP-2 localization, co-immunoprecipitation with AP2, flow cytometry for receptor surface expression, and confocal microscopy for endosome tracking. High-throughput screening campaigns for endocytosis modulators can also be supported. Researchers are encouraged to contact Ascent Research for further product details and technical support.
