This product comprises a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ATP6V0A2 gene in the human SK-HEP-1 cell line. The resulting loss-of-function model eliminates expression of the V-ATPase a2 subunit, a key component of the vacuolar ATPase V0 domain. The heterogeneous polyclonal pool contains diverse genetic edits, providing a robust system for studying ATP6V0A2 functions without the artifacts associated with single-cell cloning. This format is particularly advantageous for researchers requiring stable, population-level phenotypes for high-throughput applications or long-term studies.
The host SK-HEP-1 cell line is derived from the ascitic fluid of a patient with liver adenocarcinoma and exhibits a unique dual epithelial-endothelial phenotype. This characteristic makes it a valuable model for investigating both hepatic tumor biology and endothelial cell behavior. SK-HEP-1 cells express markers of both lineages, facilitating studies on tumor microenvironment interactions, metastasis, and endothelial transdifferentiation. The knockout of ATP6V0A2 in this context allows exploration of V-ATPase functions in a cell line relevant to cancer and vascular research.
ATP6V0A2 encodes the a2 subunit of the V0 domain of vacuolar ATPase, essential for acidifying endosomes and lysosomes. Its activity is regulated by PI3K/AKT/mTOR signaling, nutrient availability, cellular pH, and growth factors like EGF and insulin. The a2 subunit interacts with other V-ATPase subunits (e.g., ATP6V1A, ATP6V1B2), assembly factors (ATP6AP1, TMEM199), Cl- channels, and Na+/H+ exchangers. Downstream, ATP6V0A2 influences activation of lysosomal hydrolases (cathepsins), matrix metalloproteinases, collagen-processing enzymes, and glycosyltransferases. Disruption impairs cargo sorting, lysosomal enzyme maturation, and vesicular trafficking, leading to defective glycosylation and ECM remodeling.
In the SK-HEP-1 model, ATP6V0A2 knockout provides a system to dissect V-ATPase-dependent processes in cancer and endothelial contexts. The dual phenotype enables examination of how endosomal acidification affects migration, invasion, and pH signaling. Endothelial features allow studies on glycosylation-mediated cell interactions and vascular behavior. This model is relevant to human diseases like cutis laxa type IIA and wrinkly skin syndrome, characterized by aberrant glycosylation and matrix defects.
Researchers can utilize this polyclonal knockout in a variety of experimental approaches. LysoTracker staining and immunofluorescence for LAMP1 evaluate endolysosomal pH and integrity. Western blotting confirms loss of ATP6V0A2 and assesses changes in V-ATPase subunits. Functional studies may include glycosylation profiling, collagen secretion assays, and transwell migration to quantify matrix remodeling and cell motility. This model is suited for drug screening targeting V-ATPase function or glycosylation pathways, and for fundamental studies of endocytosis, vesicular trafficking, and ECM dynamics. Contact Ascent Research for further information.
