ABCG2 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited cell population featuring targeted disruption of the ABCG2 gene, eliminating expression of the ABCG2 efflux transporter (BCRP). This polyclonal pool comprises a heterogeneous mix of edited alleles, providing a robust loss-of-function model without clonal selection. The knockout was achieved via CRISPR/Cas9-mediated gene disruption, abrogating ABCG2??s ATP-dependent transport activity. This product is ideal for bulk knockout studies of drug resistance and transporter function in a hepatocyte context.
The Huh-7 cell line is a well-differentiated hepatocellular carcinoma line from a liver tumor of a 57-year-old Japanese male. Huh-7 retains hepatocyte-specific functions including cytochrome P450 expression and drug metabolism, making it a relevant model for liver biology and cancer research. Its adherent growth and stable phenotype support diverse in vitro assays such as drug sensitivity screening and transporter uptake studies.
ABCG2 extrudes xenobiotics and chemotherapeutics (e.g., mitoxantrone, topotecan) via ATP hydrolysis, limiting intracellular drug accumulation. It is transcriptionally upregulated by NRF2, AhR, HIF-1??, and PPAR??, and modulated by cytokines TNF-?? and IL-1??. Adaptor proteins PDZK1 and NHERF1 tether ABCG2 to the actin cytoskeleton, and homodimerization is required for function. Downstream, ABCG2 reduces chemotherapeutic efficacy and controls uric acid secretion and heme efflux. Knockout of ABCG2 abolishes these efflux functions, sensitizing cells to substrate drugs.
In Huh-7 cells, ABCG2 knockout directly addresses hepatic drug efflux mechanisms. Normally expressed on the canalicular membrane, ABCG2 mediates biliary excretion of drugs and metabolites. Its overexpression is linked to multidrug resistance in hepatocellular carcinoma. The knockout model enables dissection of ABCG2-specific resistance, assessment of altered drug pharmacokinetics, and evaluation of ABCG2 as a therapeutic target, while also facilitating the study of heme and urate transport in a liver-relevant system.
These polyclonal cells are suited for multidrug resistance investigations, ABC transporter functional assays, and high-throughput drug sensitivity screens. Experiments such as Hoechst 33342 efflux, mitoxantrone uptake, and MTT cytotoxicity assays directly measure transporter activity and chemosensitivity. Confirmatory analyses include RT-qPCR, western blotting, and immunofluorescence. The ready-to-use knockout pool accelerates research into ABCG2 biology, inhibitor development, and functional transporter studies in a hepatic context.
