The ATP11B Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human hepatic sinusoidal endothelial-like adenocarcinoma cell line, in which the ATP11B gene has been disrupted via CRISPR/Cas9-mediated gene targeting. This loss-of-function model enables the study of ATP11B-dependent phospholipid translocation and membrane asymmetry in a well-characterized hepatic endothelial context, without relying on single-cell clonal isolation. The knockout product format provides a heterogeneous population of edited cells, offering a robust tool for functional genomics and drug discovery applications where intact cellular heterogeneity is advantageous.
SK-HEP-1 is a human hepatic sinusoidal endothelial-like adenocarcinoma cell line originally isolated from the liver ascites of a Caucasian male. It serves as a widely accepted model for liver sinusoidal endothelial cells, displaying key phenotypic features such as scavenging receptor activity and angiogenic capacity. The line is frequently employed to investigate endothelial cell biology, hepatic clearance mechanisms, and tumor angiogenesis. Its dual epithelial and endothelial characteristics make it a versatile platform for examining hepatic pathophysiology, including liver cancer progression and drug resistance phenotypes.
ATP11B encodes a member of the P4-ATPase flippase family that actively translocates phosphatidylserine and phosphatidylethanolamine from the extracellular to the cytoplasmic leaflet of the plasma membrane, thereby maintaining phospholipid asymmetry. This flippase function is dependent on heterodimerization with accessory proteins CDC50A and CDC50B. ATP11B activity is regulated upstream by miR-34a and exerts downstream control over phosphatidylserine distribution, which in turn modulates apoptotic signaling, myoblast fusion, and vesicular trafficking pathways. Disruption of ATP11B disrupts this critical flippase complex and perturbs the spatiotemporal organization of membrane phospholipids, impacting cellular processes that rely on membrane curvature and surface exposure of phosphatidylserine.
In the SK-HEP-1 background, loss of ATP11B provides a unique model to dissect the role of phospholipid flippase activity in hepatic sinusoidal endothelial biology. This includes investigating how membrane asymmetry influences scavenging function, endothelial tube formation, and migration, which are key processes in angiogenesis and liver homeostasis. Moreover, since SK-HEP-1 exhibits both endothelial and tumorigenic properties, the knockout model facilitates the study of ATP11B??s contribution to cancer cell signaling, metastatic potential, and response to therapeutics. The interplay between flippase-mediated lipid asymmetry and drug efflux or sensitivity can be explored in this system.
Typical research applications include annexin V staining to assess phosphatidylserine externalization, fluorescent phospholipid uptake assays to monitor flippase activity, and western blotting or immunofluorescence for protein expression analysis. The model is also suitable for flow cytometry, migration and invasion assays, tube formation angiogenesis assays, and drug sensitivity profiling. These applications are particularly valuable for screening compounds that modulate phospholipid signaling or for validating the role of ATP11B in hepatic endothelial pathophysiology. For further details, please contact Ascent Research.
