The GPAT3 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the GPAT3 gene has been disrupted. This product offers a genetically defined model system for investigating the initial step of de novo glycerolipid biosynthesis. By targeting GPAT3 in the near-haploid HAP1 cell line, the polyclonal population provides a robust loss-of-function resource without requiring single-cell cloning, enabling functional analysis of GPAT3-dependent metabolic pathways directly in a human genetic screening background.
HAP1 is a near-haploid human cell line originally derived from the KBM-7 chronic myeloid leukemia lineage. Its haploid karyotype simplifies gene-editing strategies, as disruption of a single allele can result in a functional null phenotype, making it particularly useful for genetic screens and knockout validation. The HAP1 background supports applications in cancer biology, drug target identification, and functional genomics, while retaining key features of human metabolic signaling. The GPAT3 knockout in this context allows unambiguous interrogation of the gene??s role in lipid metabolism without interference from redundant copies.
GPAT3 (glycerol-3-phosphate acyltransferase 3) catalyzes the acylation of glycerol-3-phosphate with acyl-CoA to produce lysophosphatidic acid (LPA), the initial step in triacylglycerol and phospholipid biosynthesis. Transcriptionally activated by PPAR??, SREBP1c, and C/EBP?? downstream of insulin signaling, GPAT3 generates LPA that is subsequently utilized by AGPAT, lipin, and DGAT enzymes to form phosphatidic acid, diacylglycerol, and triacylglycerols, respectively. GPAT3 associates with perilipin proteins on lipid droplets and requires acyl-CoA as a cofactor. Disruption of GPAT3 thus blocks the production of key glycerolipid intermediates, profoundly altering cellular lipid homeostasis and membrane composition.
In the near-haploid HAP1 background, GPAT3 loss severely compromises de novo triacylglycerol and phospholipid synthesis, reducing lipid droplet formation and altering membrane dynamics. This polyclonal population enables studies of lipid storage, lipotoxicity, and energy metabolism in a human cell system suited to high-throughput screening. With unlimited proliferation and a stable karyotype, these GPAT3 knockout cells provide a robust platform for exploring connections between glycerolipid synthesis and insulin resistance, adipogenesis, and metabolic disease.
Typical applications include functional genomics of lipid metabolism, high-throughput drug screening for metabolic disorders (obesity, NAFLD), and investigation of insulin signaling and adipocyte differentiation. Compatible assays: western blotting, RT-qPCR, lipidomics, oil red O staining, triglyceride quantification, and metabolic flux analysis with labeled glycerol. Drug sensitivity studies using lipogenesis inhibitors can validate therapeutic targets. For inquiries, contact Ascent Research.
