The AKR1C3 Knockout HT29 Polyclonal Cells represent a heterogeneous CRISPR/Cas9-edited cell population with targeted disruption of the AKR1C3 gene in the HT29 colorectal adenocarcinoma epithelial cell line. This polyclonal knockout model is supplied as a mixed pool of edited cells, enabling researchers to study loss-of-function effects without clonal selection bias. The CRISPR/Cas9-mediated gene disruption abolishes AKR1C3 protein expression across the population, creating a versatile tool for functional genomics, drug target validation, and mechanistic studies in adenocarcinoma biology. The product is intended for research use only and is delivered with standard quality control assessments to confirm target locus editing.
Derived from a 44-year-old Caucasian female with colorectal adenocarcinoma, the HT29 cell line serves as a well-characterized model of intestinal epithelial physiology and colorectal cancer. These cells form polarized monolayers with apical junctional complexes and exhibit adherent growth in culture, recapitulating key features of colorectal adenocarcinoma epithelium. HT29 cells are widely employed in cancer biology, drug development, and toxicology due to their robust growth kinetics and responsiveness to pharmacological perturbation. The availability of AKR1C3 knockout in this background allows focused dissection of steroid and prostaglandin metabolic pathways within a clinically relevant colorectal tumor context.
AKR1C3 encodes an aldo-keto reductase that catalyzes NADPH-dependent reduction of carbonyl groups in steroid hormones and prostaglandins, functioning primarily as a 17??-hydroxysteroid dehydrogenase to convert androstenedione to testosterone and prostaglandin D2 to 9??,11??-prostaglandin F2. The enzyme is regulated by upstream signals including NRF2, IL-6, androgens, and steroidogenic factor 1 (SF1), and its activity modulates levels of downstream effectors such as testosterone, estradiol, and prostaglandin F2??, which in turn signal through androgen and estrogen receptors. NADPH serves as an essential cofactor in these reactions. Disruption of AKR1C3 in HT29 cells is expected to impair these conversions, thereby altering the balance of bioactive steroids and prostaglandins and perturbing downstream receptor-mediated signaling cascades.
In the HT29 colorectal adenocarcinoma model, loss of AKR1C3 provides a unique opportunity to investigate the convergence of steroid hormone metabolism and prostaglandin signaling in tumor epithelial cells. This knockout model may reveal how altered androgen and estrogen receptor activation, coupled with changes in prostaglandin F2?? production, influences colorectal cancer cell proliferation, differentiation, and inflammatory responses. Given HT29??s epithelial origin, the system is particularly suited for studying epithelial-mesenchymal interactions and the role of intracrine hormone metabolism in shaping the tumor microenvironment. The polyclonal nature of the knockout population preserves cellular heterogeneity, which can more accurately reflect in vivo tumor diversity than monoclonal isolates.
Researchers can employ this polyclonal knockout model in a variety of assays, including testosterone conversion enzyme assays to confirm loss of catalytic function, RT-qPCR and Western blotting for gene and protein expression analysis, cell proliferation measurements using MTS or MTT, and prostaglandin E2 ELISA to assess downstream metabolic effects. Hormone measurement via ELISA and drug cytotoxicity assays facilitate studies on chemoresistance mechanisms and endocrine disruption. Primary applications include prostate cancer drug target research, colorectal cancer endocrinology, metabolic enzyme studies, investigation of drug metabolism and chemoresistance, and analysis of inflammation and prostaglandin signaling pathways. For further technical details or custom options, please contact Ascent Research.
