The ABCC2 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the gene encoding the multidrug resistance-associated protein 2 (MRP2) has been disrupted. This loss-of-function model abolishes MRP2-mediated ATP-dependent efflux of conjugated organic anions, resulting in intracellular accumulation of substrates that are normally excreted into bile. The polyclonal nature ensures a heterogeneous pool of edited cells, providing a robust system for studying transporter biology without clonal artifacts.
The host cell line, SK-HEP-1, originates from ascites of a patient with liver adenocarcinoma and serves as a widely used in vitro model for hepatocyte function, drug metabolism, and hepatic transporter research. These epithelial cells endogenously express key uptake and efflux transporters, making them relevant for investigating hepatobiliary disposition mechanisms despite their non-hepatic origin. Their utility in transporter studies is well-documented, particularly for ABC efflux transporters.
ABCC2 encodes MRP2, an integral membrane transporter localized to the apical (canalicular) membrane of hepatocytes. Its activity is transcriptionally regulated by nuclear receptors including PXR (NR1I2), FXR (NR1H4), and CAR (NR1I3), which respond to bile acids such as chenodeoxycholic acid, as well as by the oxidative stress sensor Nrf2 (NFE2L2). Proinflammatory cytokines IL1B and TNF can suppress MRP2 expression. The transporter effluxes a broad array of substrates: bilirubin glucuronides, leukotriene C4, glutathione conjugates, glucuronidated bile acids, and drug conjugates such as methotrexate and cisplatin. MRP2 functionality is supported by interaction partners like PDZK1 (NHERF3) and ezrin/radixin/moesin proteins that stabilize its apical localization, and by ATP hydrolysis driving substrate translocation.
The knockout of ABCC2 in SK-HEP-1 cells recapitulates the transport defect underlying Dubin-Johnson syndrome, characterized by impaired biliary excretion and conjugated hyperbilirubinemia. By eliminating MRP2 efflux, the model causes intracellular retention of conjugated metabolites and xenobiotics, thereby mimicking hepatotoxicity susceptibility and chemoresistance phenotypes. It enables dissection of how loss of this transporter alters cellular pharmacokinetics and how compensatory mechanisms involving ABCC1, ABCB1, or ABCG2 may be engaged.
This knockout model is suited for a range of experimental applications, including substrate efflux assays using fluorescent probes such as CDFDA, Western blotting for MRP2 expression, RT-qPCR for ABCC2 transcript analysis, and immunofluorescence to assess transporter localization. Functional studies can employ vesicular transport assays, intracellular drug accumulation measured by flow cytometry, and ATPase activity assays. It supports drug sensitivity screening (MTT), Nrf2 pathway activation analysis, and global transcriptomic profiling (RNA-seq) to investigate transporter-mediated drug resistance, hepatobiliary toxicity, pharmacokinetic modeling, and drug-drug interactions. For additional details, please contact Ascent Research.
