The AMDHD2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the AMDHD2 gene in the HT29 human colorectal adenocarcinoma cell line. This polyclonal pool comprises a heterogeneous mixture of edited cells, each carrying distinct gene disruptions generated via CRISPR/Cas9-mediated targeting, providing a robust loss-of-function model without single-cell cloning. The product is supplied as a live cell population suitable for immediate culture and experimental use, enabling streamlined assessment of AMDHD2 function in a disease-relevant epithelial context.
The HT29 cell line, derived from a primary colorectal adenocarcinoma of a 44-year-old female, is a widely used model of intestinal epithelial biology. These cells exhibit typical epithelial morphology and can be induced to differentiate into mucin-secreting goblet cells upon treatment with agents such as methotrexate or sodium butyrate. HT29 cells are extensively employed in studies of intestinal barrier function, mucus production, and colorectal cancer pathogenesis. Their capacity for differentiation makes them particularly suited for investigating how metabolic enzyme alterations impact cellular differentiation and cancer progression.
AMDHD2 encodes a putative amidohydrolase domain-containing enzyme predicted to catalyze a hydrolytic step in amino acid or purine metabolism. Based on its domain architecture, AMDHD2 likely requires cofactors such as NAD+ or divalent metal ions for enzymatic activity. The gene’s expression is regulated by metabolic transcription factors ATF4 and NRF2, key sensors of cellular stress and nutrient status, and is responsive to oxidative stress signals. Within the histidine degradation pathway, AMDHD2 is proposed to function alongside histidine ammonia-lyase, urocanate hydratase, imidazolonepropionase, and formiminoglutamate hydrolase, ultimately contributing to the production of N-formimino-L-glutamate. Knockout of AMDHD2 in HT29 cells is hypothesized to redirect metabolic flux, potentially impacting downstream metabolites that influence energy homeostasis and redox balance.
In the HT29 colorectal cancer background, loss of AMDHD2 function is anticipated to perturb central carbon and nitrogen metabolism, creating a unique experimental platform for probing metabolic vulnerabilities specific to colorectal adenocarcinoma. Given HT29??s capacity for differentiation, this knockout model enables investigation of how AMDHD2-mediated metabolic changes intersect with epithelial differentiation programs and mucin secretion. The interplay between AMDHD2-dependent metabolism and key stress-response pathways regulated by ATF4 and NRF2 can be systematically dissected, offering insights into adaptive mechanisms that support tumor growth under metabolic stress.
This polyclonal knockout cell population is ideally suited for a broad range of applications, including functional characterization of AMDHD2 in cancer metabolism via transcriptomic (RNA-seq) and metabolomic (LC-MS) profiling, screening for small-molecule inhibitors using cell viability and proliferation assays, and exploring differentiation-dependent metabolic reprogramming through Seahorse flux analysis and flow-cytometry-based metabolic marker detection. Researchers can validate gene disruption by western blotting for AMDHD2 protein levels and RT-qPCR for mRNA expression. The model also supports co-culture and barrier function studies. For further information, please contact Ascent Research.
