The CRISPR/Cas9-edited HDAC6 Knockout SK-HEP-1 Polyclonal Cells are a genetically diverse pool of SK-HEP-1 cells with targeted disruption of the HDAC6 gene. This loss-of-function model, generated via CRISPR/Cas9-mediated gene editing, provides a population-level knockout to study HDAC6 biology while avoiding clonal selection biases. Suitable for functional genomics, these cells enable investigation of cytoplasmic deacetylase functions and microtubule-dependent processes.
The SK-HEP-1 host cell line, isolated from the ascites of a patient with liver adenocarcinoma, displays characteristics of hepatic sinusoidal endothelial cells. Such cells form the lining of liver sinusoids and are central to macromolecule exchange, leukocyte trafficking, and debris clearance. Their endothelial-like phenotype and cancer origin provide a platform for investigating liver sinusoidal function and metastatic progression.
HDAC6 is a cytoplasmic deacetylase that modulates non-histone substrates, including ??-tubulin, HSP90, cortactin, and ??-catenin. It is regulated by EGF, TGF-??, IL-6, and kinases such as Aurora A and GSK3??. Deacetylation of ??-tubulin governs microtubule dynamics, while HSP90 deacetylation affects chaperone activity. Cortactin deacetylation influences actin remodeling and migration, and ??-catenin deacetylation impacts Wnt signaling and adhesion. HDAC6 interacts with p97/VCP, ubiquitin, and 14-3-3 to coordinate protein degradation through the aggresome?Cautophagy pathway and ubiquitin-proteasome system, integrating stress responses and cell motility.
In SK-HEP-1 cells, HDAC6 knockout leads to hyperacetylation of ??-tubulin and HSP90, disrupting microtubule stability and protein folding. This impairs aggresome formation and autophagic clearance of misfolded proteins, potentially contributing to liver cell stress. Altered cortactin and ??-catenin deacetylation may affect focal adhesion and cell migration, providing a model for examining HDAC6??s role in hepatocellular carcinoma invasion and metastasis. The polyclonal knockout thus offers insights into liver sinusoidal endothelial biology and cancer progression.
Research applications include drug target validation for HDAC6 inhibitors in cancer and neurodegeneration, and studies on protein aggregation using immunofluorescence for acetylated ??-tubulin and aggresome assays. The cells support investigation of liver sinusoidal endothelial functions, immune surveillance, and metastasis via migration/invasion assays. Co-immunoprecipitation with HSP90 or p97/VCP, western blotting for acetylated HSP90 and ??-catenin, and transcriptomic analyses (RNA-seq, RT-qPCR) can dissect signaling. For further assistance, contact Ascent Research.
