The ACACA Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt acetyl-CoA carboxylase alpha (ACACA) function for metabolic and signaling research. Using CRISPR/Cas9-mediated gene disruption in a pool of HAP1 cells, this product provides a heterogeneous loss-of-function model without single-cell cloning, enabling robust pooled analyses and high-throughput applications. The polyclonal format maintains genetic diversity while effectively reducing target gene expression, making it suitable for studying ACACA-dependent processes in a near-haploid background.
HAP1 is a fibroblast-like, near-haploid cell line derived from a patient with chronic myeloid leukemia. Its adherent growth and stable karyotype simplify CRISPR/Cas9 editing and subsequent maintenance of knockout populations. The haploid genome ensures that disruption of a single allele yields functional knockout, eliminating the complexity of heterozygous mutations and facilitating unambiguous genotype-phenotype correlation. HAP1 cells are widely adopted in functional genomics, drug target screening, and metabolic studies due to their scalability and compatibility with imaging-based assays.
ACACA encodes acetyl-CoA carboxylase alpha, which catalyzes the ATP-dependent carboxylation of acetyl-CoA to malonyl-CoA, the rate-limiting step in de novo fatty acid synthesis. This enzyme is regulated by upstream signals: AMPK-mediated phosphorylation inhibits activity, allosteric activation is driven by citrate, and transcriptional induction by SREBP1c occurs downstream of insulin and glucagon. The malonyl-CoA product serves as a substrate for fatty acid synthase (FASN) and allosterically inhibits carnitine palmitoyltransferase 1 (CPT1), thereby blocking mitochondrial fatty acid oxidation. ACACA operates within a lipogenic complex involving covalent biotin attachment, FASN, and ATP citrate lyase (ACLY), positioning it as a central coordinator of lipid and carbohydrate metabolism.
In the HAP1 context, ACACA disruption abrogates malonyl-CoA synthesis, enabling direct assessment of impaired lipogenesis and its metabolic consequences. The near-haploid state permits clear phenotypic linkage to ACACA loss, and the fibroblast-like morphology is ideal for lipid droplet staining and high-content imaging. This model allows dissection of how ACACA integrates hormonal and nutrient signals??such as those from AMPK, insulin, and citrate??to control metabolic flux. Given the involvement of lipogenesis in cancer proliferation, the knockout pool serves as a platform for studying context-specific dependencies in leukemia-derived cells.
Typical research applications include modeling metabolic disorders like obesity, type 2 diabetes, and non-alcoholic fatty liver disease, where ACACA is a validated therapeutic target. Assays include radioactive acetate incorporation to measure lipogenesis, LC-MS-based malonyl-CoA quantification, and Seahorse metabolic flux analysis of mitochondrial respiration. Lipid droplet staining with BODIPY or Nile Red and RT-qPCR for lipogenic genes (e.g., FASN, SREBP1c) provide complementary readouts. The polyclonal population supports pooled CRISPR screening for modulators of lipogenic signaling. For further details, contact Ascent Research.
