The ABCC4 Knockout HEK293T Polyclonal Cells comprise a population of HEK293T cells engineered via CRISPR/Cas9-mediated gene disruption to eliminate functional expression of the ABCC4 (ATP-binding cassette subfamily C member 4) transporter. These polyclonal knockout cells provide a loss-of-function model for investigating ABCC4-dependent efflux of cyclic nucleotides and organic anions in a human cell background without reliance on pharmacological inhibitors.
The HEK293T cell line originates from human embryonic kidney cells transformed with adenovirus 5 DNA and stably expresses the SV40 large T antigen, which enhances episomal replication and facilitates high-level protein expression. This robust and well-characterized line is a standard platform for lentiviral production, receptor signaling studies, and heterologous transporter expression, making it well-suited for examining the functional consequences of ABCC4 disruption.
ABCC4 encodes an ATP-binding cassette transporter that actively extrudes cyclic nucleotides (cAMP and cGMP), nucleotides, and diverse organic anions including drug metabolites, prostaglandins, and leukotrienes. By lowering intracellular concentrations of cAMP and cGMP, ABCC4 dampens signaling through the cAMP/PKA and cGMP/PKG axes. Transcriptional regulation of ABCC4 involves factors such as NRF2, FXR, and PXR, while kinase-dependent modulation is mediated by PKA and PKG. The transporter further interacts with scaffolding proteins PDZK1 and SNX27, which influence its subcellular localization and functional coupling to downstream effectors.
In HEK293T cells, which retain endogenous cyclic nucleotide signaling machinery, ABCC4 knockout is expected to elevate baseline cAMP and cGMP levels, thereby enhancing PKA and PKG activity. This altered signaling landscape provides a controlled system to dissect ABCC4??s contribution to drug resistance, xenobiotic metabolism, and inflammatory mediator release, independent of confounding variables present in primary cells or cancer lines with complex genetic backgrounds.
Researchers can employ these ABCC4 knockout HEK293T polyclonal cells in a wide array of functional assays, including radioactive or fluorescent substrate efflux measurements to quantify transport activity, HPLC-based metabolite profiling, and cAMP/cGMP enzyme immunoassays to map cyclic nucleotide dynamics. The model supports pharmacological screening of transporter inhibitors, investigation of ABCC4-dependent resistance to anticancer agents, and co-immunoprecipitation studies with PDZK1 or SNX27 to probe protein complexes. Standard techniques such as western blotting, RT-qPCR, and immunofluorescence further enable validation of knockout efficiency and assessment of downstream signaling components. For additional details or to request a quote, please contact Ascent Research.
