Description

The SMPD3 Knockout HaCat Cell Line is a CRISPR/Cas9-edited knockout cell line with targeted disruption of the SMPD3 gene in HaCat human keratinocytes. This loss-of-function model is designed for investigating neutral sphingomyelinase 2 (nSMase2) function in skin biology and disease.

HaCat cells are a spontaneously immortalized human keratinocyte line widely used for studying epidermal differentiation, barrier function, and skin pathologies. Their stable phenotype and ease of culture make them a preferred host for genetic perturbations, and the SMPD3 knockout provides a reproducible system to examine sphingolipid signaling in keratinocytes.

SMPD3 encodes neutral sphingomyelinase 2 (nSMase2), a membrane-associated phosphodiesterase that hydrolyzes sphingomyelin into the bioactive lipid ceramide and phosphocholine. This enzymatic step is a central hub in sphingolipid metabolism, linking extracellular stimuli such as TNF-alpha, UV radiation, oxidative stress, and p53 activation to intracellular ceramide-mediated signaling. nSMase2 physically interacts with the TNF receptor and FAN to relay apoptotic signals, and its product ceramide directly promotes activation of caspase-3 and JNK while also modulating NF-kB and p38 MAPK pathways. Disruption of SMPD3 therefore eliminates nSMase2 activity, blocking stress-induced ceramide generation and attenuating downstream pro-apoptotic and inflammatory programs.

In human epidermal keratinocytes, nSMase2-derived ceramide is indispensable for programmed differentiation, cornified envelope formation, and maintenance of the skin barrier. Loss of SMPD3 in HaCat cells impedes ceramide-driven differentiation and confers resistance to ceramide-dependent apoptosis, replicating features observed in disorders characterized by impaired barrier function, such as atopic dermatitis and psoriasis. This knockout thus serves as a relevant cellular model to dissect how aberrant sphingolipid metabolism drives keratinocyte dysfunction, hyperproliferation, and inflammation, and to test therapeutic interventions targeting these pathways.

The SMPD3 Knockout HaCat Cell Line supports a diverse array of applications including ceramide quantification, annexin V apoptosis assays, western blot analysis of nSMase2, caspase-3, JNK, and NF-kB, immunofluorescence for differentiation markers, and RT-qPCR. It is suitable for drug screening, transwell barrier function tests, and integration into 3D skin equivalents. For detailed technical specifications and ordering information, please contact Ascent Research.