The ACAT1 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human A-549 lung adenocarcinoma cell line, designed for ACAT1 loss-of-function studies. This polyclonal cell pool offers a robust model for exploring gene function in a heterogeneous population, suitable for pooled screening and bulk assays.
The A-549 cell line, established from human lung adenocarcinoma, exhibits epithelial morphology and serves as a model for type II alveolar epithelial cells. Widely used in cancer biology and drug metabolism, these cells maintain key oncogenic signaling and metabolic features of lung adenocarcinoma.
ACAT1 encodes mitochondrial acetoacetyl-CoA thiolase, a homotetrameric enzyme that catalyzes the thiolysis of acetoacetyl-CoA to generate acetyl-CoA, linking ketone body metabolism, isoleucine degradation, and the mevalonate pathway. Its expression is co-regulated by transcription factors such as PPARGC1A, HNF4A, and PPARA, and is modulated by insulin and glucagon. ACAT1 functions within a network that includes HMGCS2, HMGCL, BDH1, and OXCT1, where the acetyl-CoA product serves as a substrate for the TCA cycle and as a precursor for HMG-CoA, thereby connecting fatty acid oxidation to cholesterol synthesis.
In the A-549 lung adenocarcinoma model, knockout of ACAT1 impairs the canonical ketone body utilization pathway, leading to altered acetyl-CoA compartmentalization and reduced substrate availability for mitochondrial respiration and lipid biosynthesis. This metabolic disruption mirrors aspects of beta-ketothiolase deficiency and provides a platform to dissect how cancer cells rewire metabolic networks to sustain proliferation. Researchers can investigate shifts in glutamine utilization, fatty acid oxidation, and redox homeostasis in the absence of ACAT1 activity.
These polyclonal knockout cells are well-suited for stable isotope-resolved metabolic flux analysis, Seahorse extracellular flux profiling, and enzymatic activity measurements to confirm thiolase disruption. They enable cancer metabolism studies, ketone body utilization assays, drug metabolism evaluations, and high-content screens for lipid metabolism modulators. Downstream functional effects can be assessed through cell proliferation, migration/invasion, and lipid accumulation assays, complemented by mass spectrometry for acetyl-CoA quantification. Western blotting and RT-qPCR provide molecular validation. For further technical information or to discuss customized applications, please contact Ascent Research.
