The ATG16L1 Knockout 143B Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 143B human osteosarcoma cell line, featuring disruption of the autophagy-related gene ATG16L1. This polyclonal model provides a heterogeneous pool of edited cells enabling robust loss-of-function studies without clonal selection artifacts. The product is designed for researchers investigating ATG16L1-dependent autophagy and innate immune signaling in a human bone cancer background.
The 143B cell line, a subclone of the TE85 osteosarcoma line, is a well-established model for bone tumorigenesis and metastasis. It exhibits aggressive growth and metastatic potential in vivo, making it suitable for dissecting molecular mechanisms underlying osteosarcoma progression. The ATG16L1 knockout in this context offers a powerful tool to examine autophagy??s role in cancer cell survival, stress adaptation, and metastatic behavior.
ATG16L1 encodes a core autophagy factor essential for autophagosome elongation. Mechanistically, it acts as an E3-like ligase within the ATG12-ATG5-ATG16L1 complex, directing LC3 lipidation to expand the phagophore membrane. Beyond autophagy, ATG16L1 negatively regulates NOD2/RIPK2-mediated inflammatory signaling, linking autophagic processes to innate immune responses. Upstream regulators include TFEB, FOXO3, mTORC1 inhibition, NOD2 stimulation, and starvation. Key interacting partners encompass WIPI2, FIP200, TECPR1, and the ATG12-ATG5 conjugate, while downstream events involve autophagosome maturation, xenophagy, and IL-1?? processing. Thus, this model interrogates the intersection of autophagy and inflammation.
In osteosarcoma, autophagy often supports survival under nutrient deprivation and therapeutic stress, yet its precise contributions to pathogenesis remain context-dependent. Therefore, ATG16L1 disruption in 143B cells enables systematic dissection of autophagy??s impact on tumor proliferation, apoptosis resistance, and metastatic dissemination. Moreover, because ATG16L1 bridges autophagy and NOD2/RIPK2-driven NF-??B activation, the knockout cells facilitate studies on how osteosarcoma cells modulate innate immune signaling within the tumor microenvironment.
Typical applications of these polyclonal knockout cells include autophagy flux analyses via western blotting for LC3-II/LC3-I ratio and immunofluorescence for LC3 puncta, co-immunoprecipitation assays to assess ATG12-ATG5 complex integrity, NOD2/RIPK2 signaling experiments, and autophagic degradation measurements. Additionally, they serve as a platform for investigating Crohn??s disease-relevant pathways and cancer cell adaptation to chemotherapy. For further technical information or ordering inquiries, please contact Ascent Research.
