The ABHD17B Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the ABHD17B gene in the HT29 human colorectal adenocarcinoma cell line. This product provides a heterogeneous pool of gene-edited cells, avoiding clonal selection and maintaining genetic diversity. Such polyclonal populations are well-suited for pooled functional screens and for experimental designs where population-level behavior is more representative of tumor heterogeneity.
The HT29 cell line is an adherent, epithelial colorectal adenocarcinoma model extensively utilized in cancer research. Originally established from a primary colon tumor, these cells retain key characteristics of intestinal epithelial biology and colorectal tumorigenesis. They harbor mutations in APC, p53, and other cancer-relevant genes, making them a robust system for studying oncogenic signaling, therapeutic responses, and metastatic behavior. Their ability to form polarized monolayers also facilitates investigations of barrier function and transepithelial transport.
ABHD17B functions as a protein depalmitoylase, removing palmitate from cysteine residues of substrates such as NRAS, HRAS, and PSD95, thereby regulating their membrane localization and signaling. It operates within the protein palmitoylation/depalmitoylation cycle, counterbalancing ZDHHC palmitoyltransferases. Depalmitoylation of RAS proteins modulates RAS and Wnt signaling pathways. ABHD17B interacts with substrate proteins and components like PPT1; upstream regulators are not well-characterized. Through dynamic control of palmitate turnover, ABHD17B influences the spatiotemporal activity of signaling networks.
In the context of HT29 colorectal cancer cells, disruption of ABHD17B offers a physiologically relevant model to examine the importance of depalmitoylation in oncogenic signaling. Aberrant palmitoylation can lead to mislocalization of NRAS and HRAS, altering their oncogenic output and impacting downstream pathways. This polyclonal knockout population enables systematic analysis of how loss of ABHD17B-dependent depalmitoylation influences colorectal cancer cell proliferation, migration, invasion, and sensitivity to targeted therapies.
Applications include acyl-biotin exchange assays for palmitoylation profiling, Western blotting and RT-qPCR for pathway analysis, and immunofluorescence for localization studies. Functional assays encompass proliferation, migration/invasion, and drug sensitivity screening for depalmitoylation inhibitors. This tool facilitates investigation of lipid modification in cancer biology. Contact Ascent Research for details.
