ADI1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. This product provides a loss-of-function model for studying ADI1 (acireductone dioxygenase 1), a key enzyme in the methionine salvage pathway, within a colorectal cancer context. The polyclonal population consists of a heterogeneous pool of cells, each carrying distinct CRISPR/Cas9-mediated gene disruptions at the ADI1 locus, ensuring a robust knockout phenotype without single-cell clonal selection. This format is ideal for population-level studies such as drug sensitivity assays and metabolomics analyses.
HT29 is a well-established epithelial cell line derived from a primary colorectal adenocarcinoma of a 44-year-old female. These cells retain characteristics of intestinal epithelium and are widely used as models for colorectal cancer and intestinal barrier function. HT29 cells harbor mutations in APC, TP53, and KRAS, key drivers of colorectal oncogenesis, and can undergo enterocytic differentiation under appropriate conditions, making them a versatile in vitro system for tumor biology research. In the context of ADI1 knockout, this cell line facilitates the investigation of methionine metabolism and its interplay with colorectal cancer progression.
ADI1 encodes acireductone dioxygenase, which catalyzes the oxidation of acireductone to formate and 2-keto-4-methylthiobutyrate, a methionine precursor, thereby linking polyamine synthesis to the S-adenosylmethionine (SAM) cycle. ADI1 also functions as an apoptosis regulator and binds the cytoplasmic tail of MT1-MMP (MMP14), potentially modulating cell migration and invasion. Thus, ADI1 sits at the intersection of metabolic and oncogenic signaling, with its activity influenced by methionine levels, stress stimuli, and MT1-MMP-mediated pathways, and impacting SAM, methionine, and polyamine pools.
Disruption of ADI1 in HT29 cells perturbs methionine salvage, altering SAM homeostasis and polyamine biosynthesis, which are critical for rapidly proliferating cancer cells. This metabolic vulnerability provides a platform for studying the reliance of colorectal cancer on methionine metabolism and methylation reactions. Additionally, the ADI1?CMT1-MMP interaction connects metabolic regulation to invasive behavior, enabling exploration of how loss of ADI1 affects cell migration. The polyclonal nature of the knockout population offers a heterogeneous background that simulates physiological diversity while abolishing ADI1 function.
This polyclonal knockout product supports a spectrum of research applications, including LC-MS metabolomics for methionine/SAM quantification, Western blotting and RT-qPCR for pathway analysis, and functional assays such as Boyden chamber migration/invasion, colony formation, drug sensitivity, and Annexin V apoptosis assays. Researchers can employ these cells to dissect metabolism-oncology crosstalk, uncover therapeutic targets, and investigate resistance mechanisms in colorectal cancer. For additional information, please contact Ascent Research.
