This product comprises a heterogeneous population of HAP1 cells with CRISPR/Cas9-mediated disruptions in the ACAA1 gene, creating a polyclonal loss-of-function model for peroxisomal 3-ketoacyl-CoA thiolase. The polyclonal format, produced by bulk gene editing, yields diverse loss-of-function alleles, minimizing clonal artifacts and enhancing experimental robustness for metabolic studies.
HAP1 is a near-haploid human myeloid cell line derived from the KBM-7 chronic myeloid leukemia (CML) line. Its predominantly haploid karyotype allows single-allele disruption to ablate gene function, facilitating efficient knockout generation. Retaining CML oncogenic signaling, HAP1 cells serve as a platform for haploid genetic screening and as a model for leukemia and metabolic disease research.
ACAA1 encodes the peroxisomal 3-ketoacyl-CoA thiolase that catalyzes the terminal step of fatty acid ??-oxidation, converting 3-ketoacyl-CoA into acetyl-CoA and chain-shortened acyl-CoA. This enzyme is critical for degrading very long-chain and branched-chain fatty acids. ACAA1 transcription is activated by PPARA and PPARD, and is induced by adiponectin and PPARGC1A. The protein interacts with ACOX1, HSD17B4, and SCP2 within the peroxisomal ??-oxidation machinery, and its import depends on PEX5. Acetyl-CoA generated by ACAA1 fuels the citric acid cycle, lipogenesis, and ketogenesis, while shortened acyl-CoA continues through ??-oxidation or enters bile acid synthesis.
Loss of ACAA1 in HAP1 cells blocks peroxisomal ??-oxidation, leading to accumulation of 3-ketoacyl-CoA and very long-chain fatty acids, recapitulating features of peroxisomal 3-ketoacyl-CoA thiolase deficiency (pseudo-Zellweger syndrome) and broader Zellweger spectrum disorders. The near-haploid background ensures a complete loss-of-function, aiding clear metabolic phenotyping. This model also enables investigation of the interplay between peroxisomal dysfunction and CML signaling, revealing metabolic vulnerabilities in leukemia and potential therapeutic targets.
These polyclonal knockout cells support fatty acid oxidation flux assays with radiolabeled palmitate, mass spectrometry lipidomics for very long-chain fatty acid quantification, and peroxisomal enzyme activity measurements. Knockout validation can be performed by ACAA1 immunoblotting, while peroxisomal morphology is assessed by immunofluorescence. RT-qPCR of PPAR target genes further characterizes the metabolic impact. The polyclonal population is ideal for drug screening and disease modeling. For detailed product information and inquiries, please contact Ascent Research.
