The TRPM2 Knockout HaCaT Cell Line is a CRISPR/Cas9-edited human knockout cell line in which the TRPM2 gene has been disrupted to eliminate functional TRPM2 calcium channel expression. This loss-of-function model provides a genetically defined system for investigating TRPM2-mediated signaling in an immortalized keratinocyte background.
HaCaT cells are a spontaneously immortalized, non-tumorigenic human keratinocyte line with p53 mutations, extensively used in dermatological research. They retain key characteristics of epidermal keratinocytes, including the ability to form stratified epithelia, secrete cytokines, and contribute to barrier function and inflammatory responses.
TRPM2 encodes a non-selective cation channel activated by intracellular ADP-ribose, hydrogen peroxide, and heat. Upon activation, it permits calcium influx that drives calmodulin/calcineurin/NFAT-dependent transcription and p38/JNK MAPK signaling. A central downstream consequence is NLRP3 inflammasome assembly, leading to caspase-1 activation and IL-1?? release. Upstream, PARP1 and CD38 generate ADPR from NAD+, while Fyn kinase and P2X7 receptor modulate channel activity. The NUDT9 domain mediates ADP-ribose binding and channel gating.
In HaCaT keratinocytes, TRPM2 has been linked to oxidative stress-induced apoptosis, pro-inflammatory cytokine production, and epidermal barrier disruption. Thus, knockout of TRPM2 in this line allows precise examination of its role in skin inflammation, UV damage responses, and wound healing. It offers a platform to study how TRPM2-dependent calcium signals influence keratinocyte function in diseases such as psoriasis.
This cell line supports diverse assays, including calcium imaging and ROS measurement for acute signaling, ELISA and caspase-1 activity assays for inflammasome output, and wound scratch or TEER assays for functional epithelial integrity. Western blotting, RT-qPCR, immunofluorescence, and flow cytometry for apoptosis enable comprehensive molecular phenotyping. The model is also suited for screening TRPM2-targeted compounds. For further information, contact Ascent Research.
