Description

The CYP2S1 Knockout HaCaT Cell Line is a CRISPR/Cas9-edited human keratinocyte model in which the CYP2S1 gene has been disrupted to create a stable loss-of-function system. This cell line enables precise investigation of CYP2S1-dependent metabolic and signaling pathways in an epidermal context, aiding mechanistic studies that require isogenic controls. It is suitable for a range of in vitro functional assays and comparative analyses with the parental HaCaT line.

HaCaT cells are a spontaneously immortalized, aneuploid, non-tumorigenic human keratinocyte line derived from adult skin. They maintain key characteristics of normal epidermal keratinocytes, including differentiation capacity and responsiveness to growth factors and environmental stimuli. Their robust proliferation and genetic stability in culture support reproducible, high-throughput experiments, making HaCaT a widely accepted model for studying skin physiology, xenobiotic metabolism, and oxidative stress responses.

CYP2S1 is a cytochrome P450 enzyme transcriptionally regulated by the aryl hydrocarbon receptor (AhR) upon binding of ligands such as polycyclic aromatic hydrocarbons and all-trans retinoic acid. It metabolizes all-trans retinoic acid and procarcinogens, linking AhR activation to detoxification and reactive oxygen species (ROS) production. CYP2S1 interacts with the electron donors NADPH-cytochrome P450 reductase and cytochrome b5, linking xenobiotic sensing, retinoid signaling, and redox homeostasis. Knockout of CYP2S1 thus provides a clean background to dissect these axes.

In keratinocytes, CYP2S1 loss impairs all-trans retinoic acid metabolism, modeling retinoid resistance relevant to psoriasis. Altered procarcinogen handling and ROS dysregulation may affect carcinogen susceptibility, relevant to squamous cell carcinoma. It also allows study of CYP2S1 in inflammatory skin diseases involving oxidative stress and retinoid signaling.

Applications include retinoic acid metabolism assays, CYP2S1 activity measurements, and AhR reporter assays. Standard validation employs western blotting, RT-qPCR, and DNA sequencing. Functional readouts include cell viability under oxidative stress (e.g., H2O2), intracellular ROS detection, and immunofluorescence. Global transcriptomic changes can be profiled by RNA-seq. These applications support research in skin biology, xenobiotic metabolism, drug toxicity, carcinogenesis, and oxidative stress. For further information or to discuss this product, please contact Ascent Research.