This product is a CRISPR/Cas9-edited polyclonal cell population originating from the Huh-7 human hepatocellular carcinoma line, with targeted disruption of the ABCC2 gene. The polyclonal pool ensures genetic diversity and robust representation of knockout events, facilitating functional studies to investigate the multidrug resistance-associated protein 2 (MRP2), an apical ATP-binding cassette transporter critical for hepatic organic anion efflux.
Huh-7 cells, isolated from a well-differentiated liver tumor of a Japanese male, are an established in vitro system for liver carcinoma biology, drug metabolism, and hepatobiliary transport. They retain many hepatocytic features, including the capacity to form polarized monolayers, making them an ideal model for investigating vectorial substrate movement and transporter function.
ABCC2/MRP2 is localized to the canalicular membrane and mediates the excretion of bilirubin glucuronides, glutathione conjugates, and drug metabolites. Its expression is transcriptionally controlled by nuclear receptors NR1I2 (PXR), NR1I3 (CAR), NR1H4 (FXR), and the transcription factor NFE2L2 (NRF2) in response to bile acids and oxidative stress. Functionally, MRP2 interacts with radixin (RDX) for apical anchoring and with PDZK1, requiring ATP hydrolysis for activity. It reduces intracellular drug levels and contributes to chemoresistance against agents like cisplatin and doxorubicin. Together with ABCC1, ABCB1, ABCG2, and phase II enzymes (glutathione S-transferases, UDP-glucuronosyltransferases), ABCC2 forms a detoxification network.
ABCC2 disruption in Huh-7 cells abolishes the primary canalicular efflux route, causing intracellular accumulation of MRP2 substrates. This phenotype mimics hepatobiliary transporter deficiencies such as Dubin-Johnson syndrome and offers a platform to study drug-induced liver injury, altered drug sensitivity, and compensatory transporter mechanisms, including crosstalk with other ABC exporters. The well-differentiated nature of Huh-7 cells allows the use of polarized cultures to assess directional transport deficits.
Applications include functional efflux assays with calcein-AM or CDCFDA, drug sensitivity testing (e.g., MTT), and LC-MS metabolite profiling. The knockout pool supports gene and protein expression analyses (RT-qPCR, western blot, immunofluorescence) and is valuable for screening MRP2 substrates/inhibitors, investigating drug-drug interactions, and studying bile acid transport and hyperbilirubinemia. For further inquiries, please contact Ascent Research.
