The HEXB Knockout SK-HEP-1 Polyclonal Cells product offers a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the human HEXB gene. Generated from the SK-HEP-1 host cell line via CRISPR/Cas9-mediated gene disruption, this heterogeneous pool carries loss-of-function mutations without clonal selection, preserving genetic diversity. The model supports loss-of-function studies and can serve as a starting point for single-cell cloning if required. It is ready for functional genomics, disease modeling, and drug discovery applications.
The SK-HEP-1 cell line, derived from human hepatic adenocarcinoma, displays an endothelial-like phenotype and serves as a model for liver sinusoidal endothelial cells (LSECs). These cells exhibit strong scavenging and endocytic functions, clearing modified lipoproteins, glycoconjugates, and nanoparticles. SK-HEP-1 retains endothelial marker expression and high lysosomal processing capacity, making it valuable for studying endocytic trafficking, lysosomal storage disorders, and hepatic vascular biology.
HEXB encodes the beta subunit of lysosomal ??-hexosaminidases A and B, which hydrolyze terminal N-acetyl-??-D-glucosamine residues from GM2 gangliosides, globosides, and glycoproteins. Hex A (????) requires the GM2 activator protein (GM2A) for substrate specificity, whereas Hex B (?¦?) acts on neutral substrates. CRISPR/Cas9-mediated disruption of HEXB eliminates both isoenzymes, blocking GM2 catabolism. HEXB expression is regulated by the MiT/TFE transcription factors TFEB, MITF, and TFE3, downstream of AMPK and mTOR signaling. In this model, knockout leads to lysosomal accumulation of GM2, GA2, and globosides, replicating Sandhoff disease pathology.
In SK-HEP-1 cells, HEXB knockout creates a liver sinusoidal endothelial model of GM2 gangliosidosis type II. The high endocytic activity of LSECs, combined with lysosomal glycolipid degradation defects, enables investigation of lipid accumulation, altered scavenging, and autophagy impairment. This system offers a unique platform to explore the interplay between endocytosis, lysosomal enzyme deficiency, and endothelial dysfunction in Sandhoff disease and related sphingolipidoses.
Key applications include modeling Sandhoff disease, studying lysosomal storage disorder mechanisms, and screening enzyme replacement or chaperone therapies. The cells support autophagy flux assays (LC3 turnover), lysosomal pH measurement, and immunofluorescence for LAMP1 and GM2 accumulation. Quantitative readouts such as Western blot for HEXB, ??-hexosaminidase activity, and LC-MS substrate quantification enable detailed phenotyping. The endocytic properties of SK-HEP-1 also make this model suitable for nanoparticle uptake and trafficking studies under lysosomal dysfunction. For more information, please contact Ascent Research.
