The ABHD17C Knockout Huh-7 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human hepatocellular carcinoma cell line Huh-7. This product introduces a targeted disruption of the ABHD17C gene, generating a loss-of-function model without specifying the precise editing outcome. The polyclonal format provides a heterogeneous pool of edited cells, reflecting the diversity of CRISPR-mediated gene disruptions, and is suitable for pooled functional studies.
The parental Huh-7 cell line is a well-differentiated hepatocyte-derived carcinoma cell line originally isolated from a liver tumor of a 57-year-old Japanese male in 1982. Huh-7 cells retain many features of hepatocytes and are widely used as an in vitro model for liver metabolism, hepatitis C virus (HCV) replication, and hepatocellular carcinoma (HCC) biology. They exhibit an epithelial phenotype and harbor activating mutations in the N-Ras signaling pathway, making them particularly relevant for studying Ras-driven oncogenic processes.
ABHD17C functions as a protein depalmitoylase that selectively removes palmitate groups from cysteine residues on target proteins, with N-Ras being a key substrate. Palmitoylation, mediated by acyltransferases such as ZDHHC9, anchors N-Ras to the plasma membrane, whereas ABHD17C-catalyzed depalmitoylation facilitates the release of N-Ras from membranes and its redistribution to endomembranes. This dynamic palmitoylation/depalmitoylation cycle modulates N-Ras subcellular localization and downstream signaling outputs, including the MAPK/ERK cascade. Consequently, ABHD17C acts as a regulator of N-Ras spatial organization and signal strength, influencing cell proliferation and survival.
In hepatocellular carcinoma, aberrant N-Ras signaling contributes to uncontrolled cell growth, migration, and therapeutic resistance. By eliminating ABHD17C expression, this knockout model disrupts the depalmitoylation step of the N-Ras cycle, thereby altering N-Ras steady-state localization and signaling dynamics. Given that Huh-7 cells endogenously express N-Ras and are responsive to Ras pathway inhibition, the ABHD17C knockout Huh-7 polyclonal population serves as a unique tool to dissect the contribution of depalmitoylation to HCC biology and to evaluate the impact of palmitoylation cycle disruption on drug sensitivity, particularly to multikinase inhibitors such as sorafenib.
These polyclonal knockout cells are ideally suited for a range of functional assays. Researchers can perform acyl-biotin exchange palmitoylation assays to quantify N-Ras palmitoylation levels, immunofluorescence to visualize N-Ras redistribution from the plasma membrane to internal compartments, and western blotting to assess phospho-ERK activation. Additionally, cell proliferation, migration, and drug sensitivity assays enable the study of ABHD17C-dependent phenotypic changes and therapeutic responses. For further details, including ordering information and technical support, please contact Ascent Research.
