The ABCB11 Knockout Huh-7 Polyclonal Cells product consists of a polyclonal population of Huh-7 cells engineered by CRISPR/Cas9-mediated disruption of the ABCB11 gene. This heterogeneous knockout pool provides a biologically relevant loss-of-function model for investigating the bile salt export pump (BSEP) without clonal selection artifacts, enabling studies that reflect the cellular diversity of the hepatic response.
The Huh-7 host cell line, originally established from a human hepatocellular carcinoma, retains key hepatocyte functions, including the expression of drug-metabolizing enzymes, bile acid transporters, and nuclear receptors. Its well-characterized hepatic phenotype makes it a widely used model for liver biology, drug metabolism, and toxicology studies, offering a physiologically relevant context for examining ABCB11 function.
ABCB11 encodes BSEP, an ATP-binding cassette transporter localized to the canalicular membrane that mediates the rate-limiting step in bile acid secretion. BSEP transports conjugated bile acids from hepatocytes into bile against a steep concentration gradient, a process essential for bile flow and cholesterol catabolism. Transcriptional regulation of ABCB11 is primarily driven by the farnesoid X receptor (FXR), which heterodimerizes with retinoid X receptor (RXR) and is activated by bile acid ligands. FXR signaling induces downstream targets such as the small heterodimer partner (SHP) and CYP7A1, forming a negative feedback loop that controls bile acid synthesis. BSEP functionally cooperates with canalicular transporters including ABCC2 and ABCB4, and its activity is modulated by interacting factors such as HAX-1 and scaffold proteins. Loss of BSEP function disrupts bile acid homeostasis, leading to intracellular accumulation of toxic bile acids and triggering cholestatic injury.
In the Huh-7 background, disruption of ABCB11 recreates a critical aspect of human cholestatic liver diseases such as progressive familial intrahepatic cholestasis type 2 (PFIC2) and drug-induced cholestasis. The polyclonal knockout cells retain the parental line??s hepatocyte-like characteristics, including relevant nuclear receptor expression (FXR, CAR, PXR) and intact bile acid biosynthetic machinery (CYP7A1, CYP8B1). This model therefore enables dissection of BSEP-dependent and -independent pathways regulating bile acid homeostasis and hepatotoxicity in a controlled in vitro setting.
This knockout product is suited for a broad range of experimental applications, including drug-induced liver injury (DILI) screening, functional characterization of bile acid transport using radiolabeled substrates or fluorescent analogs such as cholyl-lysyl-fluorescein, and identification of BSEP inhibitors or inducers. Researchers can also employ the polyclonal cells in ATPase activity assays, intracellular bile acid quantification under stress conditions, and immunofluorescence-based localization studies to investigate cholestatic mechanisms. The model supports co-culture systems and high-content screening approaches for evaluating hepatoprotective compounds. For further information on this product, please contact Ascent Research.
