The DMXL1 Knockout HT29 Polyclonal Cells comprise a population of HT29 colorectal adenocarcinoma cells with CRISPR/Cas9-mediated disruption of the DMXL1 gene. This polyclonal format provides a heterogeneous loss-of-function model without clonal selection, enabling robust investigation of DMXL1-dependent processes in human intestinal epithelial cells. The edited cells serve as a powerful tool for dissecting scaffold protein roles in endosomal trafficking and autophagy within a cancer context.
HT29 is a well-characterized human colorectal adenocarcinoma cell line with epithelial morphology, widely employed in cancer biology to examine signal transduction, differentiation, and therapeutic sensitivity. Originating from a primary tumor, these cells are adherent and exhibit intestinal epithelial features, making them ideal for studying endosomal and autophagy pathways in gastrointestinal malignancies. The DMXL1 knockout in HT29 cells creates a model that directly links scaffold protein dysfunction to colorectal cancer pathobiology.
DMXL1 is a scaffold protein that coordinates endosomal trafficking, lysosomal acidification, and autophagy by bridging the WASH complex and V-ATPase. It interacts with Notch1 and the Notch intracellular domain (NICD), linking Notch signaling to endosome sorting and actin dynamics. Upstream signals including NICD, mTORC1, and EGF regulate DMXL1, while downstream it promotes actin polymerization and autophagic flux. Key interacting partners are WASH1, FAM21, KIAA0196, and V-ATPase subunits. Disruption of DMXL1 impairs Notch-mediated transcription, reduces LC3-II conversion, leads to p62 accumulation, and alters actin cytoskeleton remodeling through the WASH complex.
In HT29 colorectal cancer cells, DMXL1 knockout disrupts Notch signaling, autophagy, and actin dynamics, collectively impacting tumorigenic pathways. Loss of function alters HES1 and HEY1 expression, reduces autophagic flux, and impairs cell migration and invasion. The model also holds relevance for neurodevelopmental disorders and epilepsy, reflecting DMXL1??s broader biological significance. It provides a clinically pertinent platform to study how scaffolding protein deficiencies drive disease progression.
Researchers can utilize this polyclonal knockout population for Western blotting of DMXL1, LC3-II, p62, and WASH complex members; immunofluorescence of LC3 puncta and LAMP1; RT-qPCR analysis of Notch targets such as HES1 and HEY1; flow cytometry for receptor recycling; and functional assays including migration, invasion, and drug sensitivity testing with 5-fluorouracil and oxaliplatin. Co-immunoprecipitation of Notch with WASH components and phalloidin staining further extend its utility. This model supports colorectal cancer research, autophagy studies, endosomal trafficking analysis, and Notch signaling investigation. For further information, please contact Ascent Research.
