The ANKRD13A Knockout HT29 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the ANKRD13A gene has been disrupted to generate a loss-of-function model in the human HT29 colorectal adenocarcinoma cell line. This polyclonal product retains the heterogeneous genetic background inherent to HT29 cells while abrogating ANKRD13A expression through targeted genome editing, enabling functional studies of this adaptor protein without clonal isolation artifacts.
The HT29 host cell line is derived from a primary human colorectal adenocarcinoma and exhibits epithelial morphology with adherent growth properties. HT29 cells carry well-characterized mutations in the APC and TP53 tumor suppressor genes, mimicking key genetic lesions found in colorectal cancer. As an established intestinal epithelial model, HT29 cells are widely employed in studies of colorectal cancer biology, drug response, and signal transduction, providing a physiologically relevant context for investigating EGFR pathway regulation.
ANKRD13A is an adaptor protein that recognizes ubiquitinated EGFR and facilitates its sorting into the ESCRT pathway, targeting the receptor for lysosomal degradation. Upon EGF stimulation, ligand-bound EGFR is ubiquitinated and bound by ANKRD13A, which interacts with the ESCRT-0 complex components Hrs and STAM, as well as ubiquitin. This interaction channels the receptor cargo to TSG101 and ESCRT-III, promoting lysosomal delivery and signal termination. Consequently, ANKRD13A-mediated degradation attenuates downstream MAPK/ERK and PI3K/AKT signaling. Loss of ANKRD13A impairs EGFR downregulation, leading to sustained receptor activation and potentiated downstream signaling.
In the context of HT29 cells, which harbor APC and TP53 mutations, disruption of ANKRD13A is highly relevant. EGFR signaling is frequently elevated in colorectal cancer, and sustained activation due to impaired receptor degradation can drive proliferation and resistance to cetuximab. The ANKRD13A knockout model enables dissection of how defective lysosomal EGFR clearance affects cancer cell behavior and drug response, providing insight into the interplay between tumor suppressor loss and endocytic trafficking defects.
Researchers can use these cells to investigate EGFR degradation mechanisms and sustained signaling in colorectal cancer. EGF time-course experiments with western blotting for EGFR, phospho-EGFR, AKT, and ERK quantify signal duration. Immunofluorescence enables visualization of EGFR trafficking, and bafilomycin A1 treatment confirms lysosomal accumulation. Cell proliferation and colony formation assays assess tumorigenic potential, while cetuximab sensitivity testing evaluates therapeutic responses. RT-qPCR may probe transcriptional targets. These applications make the cells a versatile tool for dissecting ANKRD13A function in colorectal cancer. For further information, please contact Ascent Research.
