The ATG16L1 Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the CAL-27 human tongue squamous cell carcinoma line. This product provides a loss-of-function model for ATG16L1, a core autophagy protein, through CRISPR/Cas9-mediated gene disruption, enabling precise investigation of autophagy-dependent processes in an oral cancer background. As a polyclonal population, it reflects average gene-editing outcomes without clonal selection.
CAL-27 is an HPV-negative, epithelial cell line established from human tongue squamous cell carcinoma. It is widely used as an in vitro model for oral cancer research, retaining key features such as adherent growth and invasive potential. Combining ATG16L1 knockout with this well-characterized cancer cell line allows direct evaluation of autophagy’s role in oral tumor biology, including cellular responses to nutrient deprivation and therapeutic stress.
ATG16L1 functions as a subunit of the ATG12-ATG5-ATG16L1 E3-like ligase complex, which catalyzes LC3-I lipidation to produce LC3-II, a critical step in autophagosome membrane elongation and closure. Its activity is modulated by upstream regulators MTOR, AMPK, ULK1, and TFEB-mediated transcription, as well as WNT signaling. Through its C-terminal WD40 domain, ATG16L1 binds NOD2 to connect autophagy with innate immune responses and bacterial clearance. Downstream effects include p62/SQSTM1 degradation, protein aggregate clearance, and xenophagic pathogen elimination. The protein interacts closely with ATG5, ATG12, LC3, and other core autophagy machinery components.
In oral squamous cell carcinoma, autophagy can exhibit both tumor-suppressive and tumor-promoting roles. ATG16L1 knockout in CAL-27 cells abolishes autophagosome formation and disrupts these dual functions, making it a powerful tool to dissect autophagy-dependent processes in tongue cancer. This model can be used to study how ATG16L1 loss influences tumor cell proliferation, migration, survival under nutrient stress, and sensitivity to chemotherapeutic agents, helping to clarify autophagy’s dichotomous contributions to cancer.
Key research applications include monitoring autophagy flux through LC3-II turnover and p62 accumulation assays, often in combination with bafilomycin A1, and visualizing autophagosomes via LC3 puncta immunofluorescence. The model supports screening of autophagy modulators, investigation of ATG16L1-dependent tumor suppression, and functional analysis of Crohn’s disease-associated ATG16L1 polymorphisms. Additional assays such as Transwell migration, invasion, and flow cytometric apoptosis detection further expand its utility in cancer research. For further details, please contact Ascent Research.
