ATP6V0A2 Knockout HAP1 Polyclonal Cells are a genetically modified cell population designed for the study of vacuolar ATPase function. This product consists of a heterogeneous pool of HAP1 cells that have undergone CRISPR/Cas9-mediated disruption of the ATP6V0A2 gene, resulting in a loss-of-function model for the a2 subunit of the V-ATPase V0 domain. The knockout format is polyclonal, ensuring a representative range of editing events across the population, making it suitable for pooled functional assays and genetic perturbation screens.
The HAP1 cell line, derived from the KBM-7 chronic myeloid leukemia line, is a near-haploid human male cell line widely employed as a model system for genetic knockout and perturbation screens. Its near-haploid karyotype simplifies the generation of knockout alleles, enabling effective CRISPR/Cas9-mediated gene disruption. HAP1 cells retain key signaling pathways and trafficking mechanisms, making them a relevant platform for studying the roles of genes such as ATP6V0A2 in cellular physiology.
ATP6V0A2 encodes the a2 subunit of the vacuolar-type H+-ATPase (V-ATPase) V0 domain, a critical component of the proton pump responsible for acidifying lysosomes, endosomes, and the Golgi apparatus. This subunit is vital for maintaining organellar pH, which regulates autophagy, endosomal trafficking, and mTORC1 signaling. The ATP6V0A2 protein forms complexes with V1 and V0 subunits and interacts with accessory proteins ATP6AP1, ATP6AP2, the Ragulator complex, and Rag GTPases, linking V-ATPase activity to amino acid sensing and mTORC1 activation. Upstream, TFEB transcriptionally regulates V-ATPase expression in response to nutrient deprivation, while mTORC1 and AMPK modulate V-ATPase assembly. Downstream, disruption of ATP6V0A2 impairs autophagic flux, reduces lysosomal hydrolase activity, and alters glycosylation patterns due to defective Golgi acidification.
In the HAP1 cell context, knockout of ATP6V0A2 recapitulates defects in organellar acidification and vesicular trafficking, providing a powerful model to dissect the molecular pathology of autosomal recessive cutis laxa type IIA (ARCL2A) and wrinkly skin syndrome. The near-haploid background minimizes genetic redundancy, allowing clear attribution of phenotypes such as impaired mTORC1 signaling, reduced autophagic clearance, and aberrant glycosylation. This model enables the study of TFEB-mediated lysosomal biogenesis, the interplay between V-ATPase and the Ragulator-Rag GTPase axis, and the impact of pH dysregulation on cellular homeostasis.
This product is suitable for a broad range of assays, including western blot analysis of V-ATPase subunits, LC3B, and p-S6K to monitor autophagy and mTORC1 activity; LysoTracker staining and immunofluorescence for LAMP1/2 to assess lysosomal abundance and morphology; flow cytometry-based measurements of lysosomal pH; and lectin blotting for glycosylation profiling. Applications extend to high-content screening for small molecules that restore lysosomal function or autophagy, and to mechanistic studies of endosomal trafficking and Wnt signaling modulation. This knockout cell pool is an essential tool for researchers investigating V-ATPase biology and its role in human disease. For further information, please contact Ascent Research.
