The ACAD10 Knockout HT29 Polyclonal Cells comprise a heterogeneous population of HT29 colorectal adenocarcinoma cells engineered via CRISPR/Cas9-mediated disruption of the ACAD10 gene. This polyclonal knockout model provides a versatile tool for studying loss-of-function effects in a mixed genetic background, reflecting the diversity inherent in tumor cell populations.
The HT29 cell line originates from a human colorectal adenocarcinoma and is characterized by its epithelial morphology. It serves as a foundational model in cancer research for investigating colorectal tumorigenesis, intestinal epithelial biology, and cellular differentiation processes.
The ACAD10 gene encodes a mitochondrial enzyme that initiates the ??-oxidation of fatty acids by catalyzing the conversion of acyl-CoA to enoyl-CoA, coupled with the reduction of flavin adenine dinucleotide. This reaction is integral to mitochondrial energy production from lipids. ACAD10 operates within a network regulated by PPARA, PPARG, and PGC-1??, and interacts with electron transfer flavoproteins (ETFA, ETFB, ETFDH) to shuttle electrons to the respiratory chain. It functions alongside other dehydrogenases such as ACADM, ACADS, and ACADVL, and its activity is interconnected with downstream components of fatty acid oxidation including ECI1, HADHA, and HADHB. Disruption of ACAD10 may thereby impair the generation of Acetyl-CoA, NADH, and FADH2, altering cellular energy homeostasis and lipid metabolism.
In the HT29 colorectal adenocarcinoma background, ACAD10 knockout provides a relevant model for exploring the intersection of mitochondrial fatty acid metabolism and cancer cell physiology. Given the reliance of rapidly proliferating tumor cells on lipid oxidation for energy and biosynthetic precursors, disruption of ACAD10 may uncover vulnerabilities related to metabolic stress and mitochondrial dysfunction. This model is particularly suited to investigate the role of fatty acid ??-oxidation in colorectal cancer progression, intestinal epithelial homeostasis, and the cellular response to nutrient deprivation.
Researchers can employ these polyclonal knockout cells in metabolic flux analyses, oxygen consumption rate measurements to assess mitochondrial function, and lipid utilization studies using labeled fatty acids. The model is amenable to drug sensitivity screens under metabolic stress, as well as migration and invasion assays to examine metastatic behavior. Additional applications include Western blotting and RT-qPCR to confirm ACAD10 ablation and evaluate compensatory pathway changes, and metabolomics profiling to map metabolic adaptations. For further information, please contact Ascent Research.
