The AKR1C3 Knockout HEK293T Polyclonal Cells represent a polyclonal knockout cell population generated via CRISPR/Cas9-mediated disruption of the AKR1C3 gene in the human HEK293T embryonic kidney epithelial cell line. This gene-edited product provides a heterogeneous cell pool with targeted loss of AKR1C3 function, enabling studies of aldo-keto reductase activity in a well-characterized cellular background.
HEK293T cells are immortalized human embryonic kidney epithelial cells that stably express the SV40 large T-antigen. This enables episomal replication of plasmids containing the SV40 origin of replication, resulting in high transfection efficiency and robust recombinant protein expression. Widely employed for viral packaging and transient overexpression, HEK293T offers a tractable and genetically manipulable system for dissecting molecular pathways.
AKR1C3, a member of the aldo-keto reductase superfamily, catalyzes the NADPH-dependent reduction of carbonyl groups in diverse substrates, including prostaglandins and ketosteroids. It converts prostaglandin D2 to 9??,11??-prostaglandin F2 and reduces androstenedione and estrone to testosterone and estradiol, respectively. Consequently, AKR1C3 modulates the balance of inflammatory mediators and active sex hormones. Its expression is regulated by transcription factors such as NRF2, PXR, CAR, and nuclear hormone receptors, while its activity influences downstream signaling through estrogen receptor, androgen receptor, and PPAR?? pathways. Representative pathway components interacting with AKR1C3 include CYP19A1, HSD17B1, PTGDS, and the ligands estradiol and testosterone.
In the HEK293T background, AKR1C3 knockout disrupts endogenous steroid hormone activation and prostaglandin metabolism, creating a loss-of-function model to investigate these processes. This model is particularly valuable for studying the regulatory roles of AKR1C3 in hormone-dependent signaling networks, given the cell line??s ease of transfection and compatibility with downstream analyses. The absence of AKR1C3 permits precise assessment of its contribution to cellular responses to exogenous stimuli, steroid biosynthesis, and drug metabolism, without confounding endogenous enzyme activity.
This knockout polyclonal cell pool is suited for a range of research applications, including hormone-dependent cancer studies (e.g., prostate and breast cancer), inflammatory disease modeling, and investigation of steroid and prostaglandin metabolism. Typical assays include Western blotting and RT-qPCR for expression analysis, LC-MS-based steroid profiling, prostaglandin quantification, ELISA measurement of estradiol and testosterone, and functional assays such as MTT proliferation and flow cytometric apoptosis analysis. The model also supports drug metabolism and toxicity screening, particularly for substrates of AKR1C3. For further inquiries, including bulk orders and custom licensing, please contact Ascent Research.
