The ATP11C Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human liver adenocarcinoma cell line, engineered for disruption of the ATP11C gene. This mixed clonal population provides a biologically relevant loss-of-function model without isolation of single-cell clones, enabling study of ATP11C-dependent processes in a hepatic endothelial-like context. The polyclonal composition captures a range of editing events while maintaining the functional heterogeneity often desired for phenotypic screening and pathway analysis.
The SK-HEP-1 host cell line originates from a human liver adenocarcinoma ascites fluid and exhibits features of liver sinusoidal endothelial cells, including expression of endothelial markers and ability to form barrier-like structures. Its dual hepatic and endothelial characteristics make it a valuable model for investigating hepatobiliary transport, transendothelial trafficking, and sinusoidal barrier function. This cell line has been widely employed to study liver-specific signaling, drug metabolism, and the molecular mechanisms underlying intrahepatic cholestasis.
ATP11C encodes a P4-ATPase flippase that translocates phosphatidylserine (PS) and phosphatidylethanolamine (PE) from the outer to the inner leaflet of the plasma membrane, thereby maintaining critical membrane asymmetry. This ATP-dependent activity is essential for proper localization of downstream effectors, including the bile salt export pump (BSEP), and is regulated upstream by bile acid-activated FXR/RXR signaling. ATP11C functions as a heterodimer with the obligatory chaperone CDC50A, and its flippase activity modulates the surface exposure of PS, a key signal in cell?Ccell interactions and membrane protein trafficking. In B cells, ATP11C supports B cell receptor (BCR) signal propagation, though in the hepatic context, its predominant role lies in coordinating phospholipid dynamics and bile acid efflux.
In SK-HEP-1 cells, disruption of ATP11C disrupts the asymmetric distribution of phospholipids, leading to altered membrane properties and impaired bile acid transporter localization??phenotypes that mimic aspects of cholestatic liver disease. The model recapitulates the molecular consequences of ATP11C deficiency observed in intrahepatic cholestasis and hemolytic anemia, providing a platform to dissect how flippase dysfunction impacts hepatic endothelial barrier integrity and bile acid homeostasis. Because SK-HEP-1 cells possess endothelioid characteristics, this knockout population allows investigation of how phospholipid flipping intersects with transcytosis and sinusoidal permeability, processes critical for liver function.
Researchers can utilize this polyclonal knockout product in a variety of advanced assays: flippase activity can be measured by lipid uptake or fluorescence-based translocation assays; externalization of PS is readily detected by annexin V binding; bile acid transport capacity may be assessed using radiolabeled or fluorescent bile acid analogues; and endothelial barrier function can be evaluated through transendothelial electrical resistance (TEER) or permeability assays. Additionally, gene expression profiling by RT-qPCR can monitor downstream targets such as FXR and BSEP. The model is suitable for drug-induced cholestasis evaluation and for screening compounds that restore membrane asymmetry or transporter localization. For further details and ordering information, please contact Ascent Research.
