The ACAT2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human ACAT2 gene in the HAP1 cell line. This format provides a diverse pool of gene-disrupted cells that uniformly lack ACAT2 function, enabling robust loss-of-function studies without clonal selection artifacts. The polyclonal nature accommodates scaled experimental designs while maintaining genetic heterogeneity akin to primary cell populations.
The host cell line, HAP1, is a near-haploid chronic myeloid leukemia line derived from the KBM-7 line and adapted to adherent growth. Its predominantly haploid karyotype means that a single CRISPR-induced lesion can produce a functional knockout, bypassing the need for biallelic targeting. This genetic simplicity underlies the widespread use of HAP1 in functional genomics screens, particularly for metabolic pathways where gene copy number can influence phenotype.
ACAT2 encodes acetyl-CoA acetyltransferase 2, which catalyzes the condensation of two acetyl-CoA molecules to form acetoacetyl-CoA. This reaction represents an essential branch point in metabolism, feeding both the mevalonate pathway for cholesterol biosynthesis and the ketogenic pathway that yields acetoacetate and ??-hydroxybutyrate. ACAT2 is regulated by the sterol-sensing transcription factor SREBF2 and the fatty acid?Cactivated receptor PPARA. It interacts with ACAT1 and lies upstream of HMG-CoA synthase and HMG-CoA reductase; its product, acetoacetyl-CoA, is converted to HMG-CoA, then mevalonate, squalene, and ultimately cholesterol. Consequently, ACAT2 disruption blocks this early step, perturbing lipid and ketone body homeostasis.
Within the HAP1 cellular context, ACAT2 knockout allows precise dissection of acetyl-CoA flux because the haploid genome eliminates confounding allelic effects. HAP1 cells express key regulators such as SREBF2 and PPARA, making them inherently responsive to sterol and fatty acid cues. Loss of ACAT2 is expected to reduce levels of intermediate metabolites like acetoacetyl-CoA and HMG-CoA, while potentially diverting acetyl-CoA toward fatty acid elongation or butanoate metabolism. This configuration facilitates investigation of metabolic reprogramming and feedback mechanisms, with the polyclonal population providing reproducible results across multiple independent experiments.
This knockout product is applied in lipid metabolism research, cholesterol biosynthesis studies, and drug discovery. Representative assays include Western blotting and RT-qPCR for target validation, lipidomics profiling to quantify cholesterol esters and ketone bodies, Seahorse metabolic flux analysis for mitochondrial function, and RNA-seq for transcriptomic characterization. The cells are used to model atherosclerosis, hyperlipidemia, non-alcoholic fatty liver disease, and inborn errors of ketone body metabolism. Functional genomics screens and compound testing for lipid disorders further leverage this robust loss-of-function system. For additional technical information or custom requests, please contact Ascent Research.
