The ATG16L1 Knockout DLD-1 Polyclonal Cells are a CRISPR/Cas9-generated polyclonal knockout population of the DLD-1 human colorectal adenocarcinoma cell line. This product offers a heterogeneous loss-of-function model for investigating ATG16L1-dependent autophagy and its crosstalk with inflammatory pathways. By disrupting the ATG16L1 gene in an epithelial colorectal cancer background, these cells enable population-level studies of autophagy deficiency, stress responses, and therapeutic vulnerabilities.
Originating from a Dukes?? type C colorectal carcinoma, DLD-1 cells are a widely used epithelial model for dissecting oncogenic signaling, tumor?Cstroma interactions, and drug resistance. Their adenocarcinoma phenotype provides a physiologically relevant platform to examine the role of autophagy in colorectal cancer progression, making them ideal for ATG16L1 knockout studies.
ATG16L1 is a core autophagy component forming the ATG12?CATG5?CATG16L1 E3-like ligase complex that conjugates LC3-I to phosphatidylethanolamine, generating LC3-II on autophagosomal membranes. This lipidation step is essential for autophagosome elongation and selective cargo degradation. ATG16L1 activity is modulated by upstream nutrient sensors mTORC1 and AMPK, and its interactions with NOD2 and WIPI2 link autophagy to innate immune signaling. Loss of ATG16L1 disrupts LC3-II conversion and p62/SQSTM1 clearance, while the Crohn??s disease-associated T300A variant impairs these functions and compromises bacterial handling.
In colorectal cancer, autophagy plays dual roles??supporting tumor cell survival under metabolic stress while also restraining inflammation-driven tumorigenesis. The ATG16L1 knockout DLD-1 model allows direct interrogation of these functions, particularly the autophagy?CNOD2?CRIPK2 signaling axis. By abrogating canonical autophagy, these cells can be used to probe mechanisms of chemoresistance, inflammatory cytokine regulation, and bacterial clearance in a colorectal cancer context relevant to inflammatory bowel disease-associated malignancy.
Researchers can employ standard autophagy assays, including LC3 immunoblotting, immunofluorescence puncta analysis, and flux measurements with bafilomycin A1, to confirm functional disruption. Cell viability under nutrient deprivation or drug challenge, gentamicin protection assays for intracellular bacterial survival, and co-immunoprecipitation of ATG16L1 complex members further expand the model??s utility. Phospho-signaling profiling and drug sensitivity screens can uncover compensatory pathways and therapeutic targets in autophagy-deficient colorectal cancer cells. For detailed information, contact Ascent Research.
