The Npc1l1 Knockout Hepa 1-6 Cell Line is a validated CRISPR/Cas9-edited knockout model derived from the murine Hepa 1-6 hepatoma line. This product features targeted disruption of the Npc1l1 gene, resulting in loss of Npc1l1 protein expression and function. The cell line serves as a defined genetic model for investigating hepatic cholesterol metabolism in the absence of canonical NPC1L1-mediated transport, providing a controlled system for studying liver-specific lipid handling pathways.
The parental Hepa 1-6 cell line is an established mouse hepatocellular carcinoma model originally derived from a BW7756 tumor in C57L/J mice. It retains key hepatocyte-specific functions, including de novo lipogenesis, bile acid synthesis, and lipid secretion, making it an ideal host for studying metabolic liver diseases. The sustained expression of relevant transcriptional regulators, such as SREBP2 and LXR??, in these cells supports the functional interrogation of cholesterol-responsive gene programs.
Npc1l1 encodes the Niemann-Pick C1-like 1 protein, which localizes to the hepatocyte canalicular membrane and mediates cholesterol absorption from bile. It binds cholesterol-rich micelles and facilitates internalization via clathrin-mediated endocytosis, interacting with the AP2 complex, flotillins, and Rab proteins. Npc1l1 expression is transcriptionally regulated by SREBP2 and LXR??, while its activity modulates intracellular cholesterol levels and downstream targets including CYP7A1. This positions Npc1l1 as a critical regulator of the enterohepatic circulation and cholesterol homeostasis.
In the context of hepatocellular carcinoma, Npc1l1 disruption provides a powerful tool for modeling dysregulated cholesterol absorption and biliary reabsorption. Loss of Npc1l1 function is known to attenuate diet-induced hypercholesterolemia and hepatic steatosis, making this knockout line highly relevant for non-alcoholic fatty liver disease and atherosclerosis research. The model enables precise separation of hepatic NPC1L1-dependent effects from the well-characterized intestinal role of NPC1L1, offering unique insights into liver-specific lipidology.
Researchers can utilize this knockout cell line for radiolabeled or fluorescent (NBD-cholesterol) cholesterol uptake assays to quantify transporter activity. It is ideally suited for NPC1L1 inhibitor screening, most notably for testing ezetimibe efficacy and mechanisms of action. Additional applications include Oil Red O staining to assess neutral lipid accumulation, RT-qPCR for profiling downstream targets such as Cyp7a1 and Srebp2, Western blotting for NPC1L1 and SREBP2 protein analysis, and mass spectrometry for bile acid profiling. For further technical specifications and ordering information, please contact Ascent Research.
