The ABCC1 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the ABCC1 gene has been disrupted to abolish expression of the multidrug resistance protein 1 (MRP1). This polyclonal product format was generated through genome editing of the HEK293T host cell line, yielding a heterogeneous pool of edited cells that collectively provide a loss-of-function model without the bottlenecks of single?cell clone selection. The knockout strategy eliminates MRP1 transporter activity, enabling robust investigation of organic anion, glutathione conjugate, and xenobiotic efflux mechanisms in a well?characterized cellular background.
HEK293T cells are an immortalized human embryonic kidney epithelial line that stably expresses the SV40 large T antigen, originally derived by transforming HEK293 cells with sheared adenovirus type 5 DNA. This cell line is renowned for its high transfection efficiency, rapid proliferation, and capacity for recombinant protein production and viral vector packaging, making it a preferred host for transporter studies. The epithelial origin of HEK293T cells provides a physiologically relevant context for studying membrane efflux pumps, while the T antigen facilitates episomal replication of plasmids containing the SV40 origin, further enhancing experimental throughput.
ABCC1 encodes MRP1, an ATP-binding cassette transporter that actively effluxes a broad spectrum of substrates including organic anions, leukotriene C4 (LTC4), and glutathione (GSH) conjugates of chemotherapeutic agents such as doxorubicin and vincristine. MRP1 function is transcriptionally regulated by upstream factors NRF2, p53, and HIF-1?? in response to oxidative stress and xenobiotic exposure. It operates within the NRF2/KEAP1 antioxidant pathway and cooperates with glutathione S-transferases to conjugate and eliminate toxic compounds. Downstream, MRP1 mediates cellular resistance by reducing intracellular drug accumulation, directly impacting leukotriene C4 transport and glutathione conjugate extrusion, thereby modulating inflammatory and chemoprotective responses.
In HEK293T cells, disruption of ABCC1 eliminates MRP1-mediated efflux, leading to enhanced intracellular retention of fluorescent substrates like calcein-AM and chemotherapeutics such as doxorubicin. This creates a sensitized background where dose?dependent substrate accumulation and cytotoxicity can be precisely quantified, offering a clean model to dissect MRP1 pharmacology without interference from other major ABC transporters commonly upregulated in cancer cells. The polyclonal nature of the knockout population mitigates clonal artifacts and provides a more faithful representation of heterogeneous cellular responses, which is particularly valuable for inhibitor screening and drug resistance reversal studies.
This knockout cell model supports a wide range of research applications, including multidrug resistance mechanism studies, drug efflux assay development, transporter substrate identification, and pharmacological inhibition screening. Representative assays include Western blotting for MRP1 expression, RT-qPCR for ABCC1 transcript analysis, calcein-AM efflux and doxorubicin accumulation assays to monitor transport activity, MTT-based drug sensitivity testing, and LC-MS/MS analysis of glutathione conjugates. By enabling quantitative dissection of MRP1 function in a tractable HEK293T system, this product is well suited for cancer biology research and early-stage drug discovery. For further technical details, please contact Ascent Research.
