The DNAJC13 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human colorectal adenocarcinoma HT29 cell line. This product provides a loss-of-function model for studying the role of DNAJC13, a J-domain co-chaperone involved in endosomal trafficking pathways. The polyclonal knockout pool introduces genetic heterogeneity and enables robust population-level analyses of gene disruption effects, offering a versatile tool for functional genomics and drug discovery applications.
The HT29 cell line is a widely employed in vitro model of human intestinal epithelium and colorectal adenocarcinoma. Originally isolated from a primary tumor, HT29 cells retain characteristics of enterocytic differentiation and are commonly utilized in cancer biology, signal transduction, and intestinal barrier function studies. Their epithelial origin and well-characterized signaling networks make them particularly suitable for investigating endosomal trafficking and Wnt pathway dynamics in a colorectal cancer context.
DNAJC13 encodes a co-chaperone that interacts with HSPA8 and the retromer complex, including VPS35, VPS26, and VPS29, to orchestrate retromer-dependent retrograde transport from endosomes to the trans-Golgi network. This activity is essential for the proper trafficking of cargo such as the Wnt chaperone Wntless, which is regulated by upstream signals including Rab7, SNX1, and PI3P. DNAJC13-mediated sorting influences Wnt signaling by controlling Wntless availability, thereby modulating ??-catenin stabilization and TCF/LEF transcription factor activity. Additionally, DNAJC13 intersects with autophagy-lysosomal degradation pathways, impacting protein homeostasis. Its disruption perturbs endocytic recycling and retromer-dependent processes, potentially altering downstream transcriptional programs and cellular responses.
In HT29 colorectal cancer cells, the Wnt pathway is frequently deregulated and drives tumorigenic properties such as proliferation and invasion. DNAJC13 knockout in this background disrupts retromer-mediated Wntless trafficking, leading to attenuated Wnt ligand secretion and reduced ??-catenin-dependent transcription. Consequently, this model enables dissection of the crosstalk between endosomal sorting and oncogenic signaling. It also provides a platform to explore how retromer dysfunction contributes to colorectal cancer pathophysiology and to evaluate therapeutic strategies targeting Wnt-driven growth.
Researchers can employ this polyclonal knockout cell population across diverse assays, including western blotting and RT-qPCR to monitor DNAJC13 and Wnt target gene expression, immunofluorescence to visualize retromer complex mislocalization, and co-immunoprecipitation to probe protein interactions. Functional studies such as colony formation, proliferation, wound healing, and invasion assays allow assessment of tumorigenic behavior, while flow cytometry can quantify cell surface receptor changes. The model is also applicable to drug screening for Wnt inhibitors and to mechanistic investigations of endosomal sorting defects relevant to Parkinson??s disease. For further details or technical support, please contact Ascent Research.
