The ALDH5A1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the human colorectal adenocarcinoma HT29 cell line, featuring targeted disruption of the ALDH5A1 gene. This loss-of-function model enables detailed investigation of succinate semialdehyde dehydrogenase (SSADH) function in an epithelial cancer context. The polyclonal format provides a heterogeneous knockout population ideal for functional studies without clonal selection.
The HT29 cell line, isolated from a primary colorectal tumor of a white female patient, is a widely used model of intestinal epithelium and colorectal adenocarcinoma. Its epithelial morphology and retention of colonic features make it suitable for studies of epithelial biology, oncogenic signaling, and metabolic pathways relevant to colorectal cancer.
ALDH5A1 encodes the mitochondrial enzyme SSADH, which irreversibly oxidizes succinate semialdehyde to succinate using NAD+ as a cofactor. This reaction bridges GABA catabolism to the TCA cycle: GABA is first transaminated by GABA transaminase to succinate semialdehyde, which then enters the SSADH reaction. Genetic disruption of ALDH5A1 leads to accumulation of succinate semialdehyde and its derivative 4-hydroxybutyric acid, while reducing succinate production and potentially altering redox homeostasis.
In HT29 colorectal cancer cells, ALDH5A1 knockout provides a valuable model to study how GABA shunt activity impacts mitochondrial metabolism and cancer cell physiology. Disruption of this pathway can lead to metabolic reprogramming, affecting energy production, proliferation, and oxidative stress responses. This model is also relevant for succinic semialdehyde dehydrogenase deficiency (SSADH deficiency), a neurometabolic disorder, and for exploring the role of GABA metabolism in colorectal cancer.
This knockout cell model supports diverse applications including metabolic flux analysis by LC-MS metabolomics, SSADH enzyme activity assays, gene expression analysis via RT-qPCR and Western blotting, and functional studies such as cell viability assays under metabolic stress. Redox status and mitochondrial function can be evaluated using fluorescent probes. Immunofluorescence can assess mitochondrial morphology and enzyme localization. For further information, please contact Ascent Research.
