The ABHD17C Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the ABHD17C gene in the human HT29 colorectal adenocarcinoma line. This polyclonal population provides a heterogeneous loss-of-function model suitable for studying ABHD17C functions without clonal bias, generated through CRISPR/Cas9-mediated gene disruption.
The HT29 cell line, derived from a colorectal adenocarcinoma of a 44-year-old female, displays adherent epithelial morphology and retains intestinal epithelial characteristics including absorptive capability, barrier formation, and mucus secretion. Widely used in colorectal cancer research, HT29 cells offer a relevant physiological context for investigating tumor-associated genes and signaling pathways.
ABHD17C functions as a depalmitoylase, removing S-palmitate modifications from cysteine residues on target proteins, thus regulating their membrane localization and trafficking. A key substrate is the small GTPase N-Ras; ABHD17C-mediated depalmitoylation governs N-Ras cycling between cellular membranes, which is essential for proper MAPK/ERK signal propagation. Loss of ABHD17C leads to hyperpalmitoylation of N-Ras, disrupting its subcellular distribution and causing aberrant activation of the BRAF?CMEK1/2?CERK1/2 signaling axis. This depalmitoylation cycle also interconnects with Wnt/??-catenin signaling, a pathway frequently altered in colorectal cancer.
In the HT29 background, which harbors a BRAFV600E mutation driving constitutive MEK?CERK activity, ABHD17C knockout enables the dissection of palmitoylation-dependent N-Ras contributions to signal amplitude and cancer cell behavior. The model is particularly relevant for examining how altered N-Ras localization affects colorectal cancer cell proliferation, survival, and metastatic potential.
Key applications include monitoring phospho-ERK by western blot, imaging N-Ras membrane localization via immunofluorescence, quantifying downstream gene expression by RT-qPCR, and assessing proliferation, migration, invasion, and apoptosis. These polyclonal knockout cells are also valuable for screening depalmitoylation inhibitors and studying metastasis mechanisms. For further information or technical support, please contact Ascent Research.
