The ALDH3A2 Knockout HT29 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ALDH3A2 gene, enabling loss-of-function studies in a human colorectal adenocarcinoma background. This product consists of a heterogeneous pool of HT29 cells that have undergone CRISPR/Cas9-mediated gene disruption of ALDH3A2, serving as a versatile model for investigating fatty aldehyde metabolism and its roles in cancer biology and oxidative stress responses.
The host cell line HT29 originates from a 44-year-old female with colorectal adenocarcinoma and exhibits an adherent, epithelial phenotype. These cells form a heterogeneous population capable of spontaneous differentiation, reflecting key features of intestinal epithelium. Their well-characterized signaling pathways and tumorigenic properties make HT29 cells a widely used platform for colorectal cancer research, including studies on cell proliferation, differentiation, and lipid metabolism.
ALDH3A2 encodes a fatty aldehyde dehydrogenase that catalyzes the NAD+-dependent oxidation of long-chain fatty aldehydes to corresponding fatty acids, a critical step in fatty aldehyde detoxification and lipid homeostasis. Its expression is regulated by upstream factors such as PPAR?? and the oxidative stress-responsive transcription factor NFE2L2 (Nrf2). Downstream, the generated fatty acids are activated to fatty acyl-CoA by fatty acyl-CoA synthetase and channeled into triglyceride synthesis or lipid mediator production. The enzyme functions within a network that includes sphingosine-1-phosphate lyase and fatty alcohol oxidase, preventing accumulation of cytotoxic aldehydes and subsequent oxidative damage.
Knockout of ALDH3A2 in HT29 cells impairs fatty aldehyde clearance, leading to elevated aldehyde levels and increased oxidative stress. This disruption likely alters lipid metabolic flux, affecting membrane integrity, energy storage, and signaling lipid pools. The model offers a relevant system to examine how defective fatty aldehyde metabolism impacts colorectal cancer cell behavior, including proliferation, survival under stress, and metastatic potential, and may shed light on the metabolic adaptations of tumor cells.
This polyclonal knockout population is well-suited for Sj?gren-Larsson syndrome modeling, given the syndrome’s link to ALDH3A2 mutations, and for studying fatty aldehyde metabolism in cancer drug detoxification and oxidative stress resistance. Researchers can employ assays such as western blotting and RT-qPCR to confirm expression changes, fatty aldehyde LC-MS for metabolite profiling, aldehyde dehydrogenase activity measurements, and cell viability tests under oxidative challenge. Additional readouts include lipid droplet staining and migration/invasion assays to assess phenotypic consequences. For further details, please contact Ascent Research.
