The ATXN3 Knockout SK-HEP-1 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal population of SK-HEP-1 cells with targeted disruption of the ATXN3 gene. This loss-of-function model ablates endogenous ataxin-3 expression, creating a versatile tool for investigating deubiquitinase-dependent processes in a human hepatic adenocarcinoma background. The polyclonal format ensures heterogeneity reflective of the editing pool, suitable for bulk functional studies without clonal artifacts.
The SK-HEP-1 cell line originates from the ascitic fluid of a patient with hepatic adenocarcinoma and displays epithelial characteristics. Widely employed in hepatic cancer research, these cells provide a relevant context for studying tumor biology, metabolism, and stress responses. Their robust growth and tractability make them suitable for genome engineering and subsequent functional assays.
ATXN3 encodes the deubiquitinating enzyme ataxin-3, which hydrolyzes ubiquitin chains from substrate proteins, thereby regulating proteasomal degradation, autophagy, and endoplasmic reticulum-associated degradation (ERAD). It is activated under proteotoxic stress, heat shock, proteasome inhibition, and oxidative stress. Ataxin-3 interacts with VCP/p97, ubiquitin, hHR23, Rad23, and proteasome subunits, and modulates the stability of polyubiquitinated proteins, p53, and NF-??B. Through these interactions, ATXN3 influences apoptosis and transcriptional control, positioning it at a nexus of protein quality control and stress signaling.
Disruption of ATXN3 in SK-HEP-1 cells eliminates ataxin-3??s deubiquitinase activity, resulting in accumulation of ubiquitinated species and impaired clearance of misfolded proteins. This perturbation can dysregulate p53 and NF-??B pathways, potentially altering apoptosis sensitivity and transcriptional programs relevant to hepatic tumor biology. The model thus enables dissection of how UPS dysfunction contributes to cancer cell survival and stress adaptation.
Researchers can employ this knockout model to explore mechanisms of protein aggregation, neurodegeneration-related pathways (in a non-neuronal context), and cancer cell stress responses. It is suitable for drug screening studies targeting ataxin-3 for spinocerebellar ataxia type 3, as well as for evaluating proteasome inhibitor efficacy. Typical assays include western blotting for ATXN3 and ubiquitin, proteasome activity measurements, apoptosis analysis, RT-qPCR for transcript confirmation, immunofluorescence for aggregate formation, and co-immunoprecipitation of ubiquitinated proteins. For further information, please contact Ascent Research.
