The ABHD5 Knockout HAP1 Polyclonal Cells comprise a genetically heterogeneous population of HAP1 cells in which the ABHD5 gene has been disrupted via CRISPR/Cas9-mediated gene editing. This polyclonal knockout cell population provides a robust loss-of-function model for investigating ABHD5 biology, without selection for a single clonal genotype. The product is delivered as a pool of edited cells, offering researchers a convenient and scalable tool for functional genomics and drug discovery applications related to lipolysis and lipid metabolism.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) cell line, originating from a male patient. Its haploid nature simplifies genetic manipulation and phenotypic analysis, making it a favored model for knockout studies. HAP1 cells retain key signaling pathways and metabolic features, and their use in leukemia research and functional genomics is well-established.
ABHD5 (??/?? hydrolase domain-containing protein 5) functions as an essential co-activator of adipose triglyceride lipase (ATGL, PNPLA2), the rate-limiting enzyme for triglyceride hydrolysis. It directly binds ATGL, promoting the initial step of lipolysis on lipid droplets. This interaction is regulated by perilipin 1 (PLIN1), which sequesters ABHD5 basally; upon stimulation, PLIN1 phosphorylation releases ABHD5 to activate ATGL. Additional binding partners include PLIN5, G0S2, and ABHD4. Upstream regulators such as insulin and AMPK modulate ABHD5 activity, while downstream outputs include free fatty acid and glycerol release, diacylglycerol metabolism, and PPAR?? signaling, linking lipolysis to fatty acid oxidation and metabolic gene transcription. Deficiency in ABHD5 causes Chanarin-Dorfman syndrome, characterized by systemic triglyceride accumulation.
In the HAP1 cell model, ABHD5 knockout blocks ATGL-mediated lipolysis, causing abnormal lipid droplet accumulation and disrupted triglyceride homeostasis. The near-haploid genome of these polyclonal knockout cells provides a genetically clean background for studying lipolysis and lipid droplet dynamics. This model is ideal for investigating the regulatory interplay between ABHD5, perilipins, and ATGL, and for assessing downstream effects on fatty acid oxidation and PPAR??-driven transcription. The absence of ABHD5 recapitulates cellular defects seen in Chanarin-Dorfman syndrome and obesity-related lipid storage disorders, offering a versatile platform for metabolic disease research.
These polyclonal knockout cells support a broad range of experimental approaches, including ATGL activity assays for direct lipase measurement, quantitative triglyceride analysis, and BODIPY-based lipid droplet staining to monitor lipid accumulation. They enable co-immunoprecipitation studies of ABHD5-ATGL interaction and western blotting for key lipolytic proteins such as HSL and phosphorylated perilipins. Additional applications include fatty acid release assays and PPAR?? reporter assays to evaluate downstream signaling. This model also facilitates high-throughput screening for modulators of lipolysis and lipid storage. Researchers are invited to contact Ascent Research for further information and ordering details.
