The ANKZF1 Knouckout HT29 Polyclonal Cells provide a stable CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 colorectal adenocarcinoma cell line, targeting the ANKZF1 gene. This heterogeneous pool of gene-disrupted cells enables functional studies in an intestinal epithelial background, allowing robust analysis of ANKZF1 loss-of-function effects without clonal selection.
The HT29 cell line is an adherent human colorectal adenocarcinoma line derived from a female donor, widely used as a model for intestinal epithelial biology and colon carcinoma research. Its well-characterized epithelial phenotype and tumorigenic properties provide a relevant platform for investigating gene function in cancer.
ANKZF1 encodes a mitochondrial ribosome quality control factor with peptidyl-tRNA hydrolase activity that resolves stalled mitoribosomes by releasing nascent chains from peptidyl-tRNA. This activity is activated by mitochondrial stress and the integrated stress response, where ATF4 transcriptionally upregulates ANKZF1. ANKZF1 functions downstream of these stress signals to interact with mitochondrial ribosomal proteins and the mitoribosome, facilitating the release of stalled peptides and maintaining mitochondrial translation fidelity. Its action directly impacts the synthesis of OXPHOS complex subunits, preserving mitochondrial protein homeostasis and oxidative phosphorylation capacity.
In HT29 colorectal cancer cells, ANKZF1 disruption impairs mitochondrial ribosome quality control, potentially leading to the accumulation of stalled mitoribosomes, diminished synthesis of OXPHOS subunits, and mitochondrial dysfunction. Given the elevated metabolic demands of tumor cells, this model is instrumental for studying how mitochondrial proteostasis defects influence cancer cell viability, metabolic reprogramming, and stress responses. It provides a physiologically relevant system to explore the links between mitochondrial translation, ribosome stalling, and oncogenic phenotypes in colon carcinoma.
Researchers can apply this knockout model in assays such as western blotting of OXPHOS subunits, Seahorse mitochondrial stress tests, mitochondrial ribosome profiling, and puromycin incorporation to assess mitochondrial translation. Additional readouts include RT-qPCR of mitochondrial-encoded genes and apoptosis or viability assays under stress conditions. These tools support investigations into mitochondrial translation fidelity, ribosome stalling pathways, mitochondrial dysfunction in cancer, and ATF4-mediated stress response mechanisms. For further technical information, please contact Ascent Research.
