The AKR1C2 Knockout HEK293T Polyclonal Cells are a polyclonal knockout cell population generated by CRISPR/Cas9-mediated disruption of the AKR1C2 gene in the HEK293T cell line. This loss-of-function model enables researchers to study the roles of aldo-keto reductase family 1 member C2 in steroid hormone metabolism, prostaglandin signaling, and xenobiotic detoxification. The polyclonal format provides a heterogeneous pool of knockout cells, reducing clonal artifacts and supporting robust functional analyses. AKR1C2 protein ablation can be confirmed by western blotting, ensuring a reliable knockout phenotype.
HEK293T is a derivative of the human embryonic kidney HEK293 cell line that stably expresses the SV40 large T antigen, which enhances episomal plasmid replication and enables high-level transient protein expression. This adherent cell line is widely used for protein production, virus packaging, and cell-based assays. Its renal epithelial origin offers a suitable context for investigating steroid and xenobiotic metabolism, particularly since HEK293T cells possess limited endogenous steroidogenic machinery, allowing for controlled dissection of AKR1C2 function when supplemented with exogenous substrates.
AKR1C2 is an NADPH-dependent reductase that acts on carbonyl groups of steroids, prostaglandins, and xenobiotics. It converts progesterone to 20??-hydroxyprogesterone, attenuating progesterone receptor (PGR) signaling, and similarly inactivates androgens to modulate androgen receptor (AR) activity. The enzyme also participates in retinoic acid biosynthesis. AKR1C2 is transcriptionally regulated by NRF2 and induced by IL-1?? and TNF??, linking it to oxidative stress responses. It functionally interacts with cofactor NADPH, substrates like prostaglandin E2, and steroidogenic CYP enzymes (e.g., CYP17A1, CYP19A1). Downstream, it influences prostaglandin signaling via PTGS2 and retinoic acid pathways involving RDH10 and ALDH1A1.
Knockout of AKR1C2 in HEK293T cells disrupts progesterone and androgen catabolism, potentially altering steroid receptor signaling and cellular responses to oxidative stress. This model allows for detailed examination of enzyme-substrate specificity by adding defined steroids or prostaglandins to culture medium. Because HEK293T cells can be transfected with nuclear receptors, the impact of AKR1C2 loss on AR and PGR transcriptional activity can be directly assessed. Moreover, the NRF2-AKR1C2 axis can be probed under pro-oxidant conditions to evaluate the enzyme??s role in cytoprotection and detoxification.
Applications of these polyclonal knockout cells span steroid hormone profiling by LC-MS/MS, androgen metabolism assays, and prostaglandin E2 ELISA. They are instrumental in cancer research, particularly for hormone-sensitive breast and prostate cancers, where drug resistance mechanisms can be examined using chemosensitivity assays and receptor signaling readouts. The model supports xenobiotic metabolism studies with ROS measurements and immunofluorescence. Complementation experiments can validate AKR1C2 function, and the cells are suitable for inhibitor screening. For further product details, please contact Ascent Research.
