ATG3 Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human 143B osteosarcoma line. This product offers a genetically disrupted ATG3 locus across a heterogeneous pool, enabling loss-of-function studies while avoiding clonal bias. The polyclonal format ensures robust representation of knockout phenotypes, supporting reproducible autophagy research.
Parental 143B cells are a widely used osteosarcoma model with osteoblast-like characteristics, originally isolated from a primary bone tumor. They maintain osteoblastic marker expression and are highly tumorigenic and metastatic in vivo, making them particularly relevant for bone cancer and bone metabolism studies. The cell line??s stable genomic background and ease of culture facilitate high-efficiency genome editing.
ATG3 functions as an E2-like conjugating enzyme in the conserved autophagy pathway. It acts downstream of the E1-like enzyme ATG7 to catalyze the conjugation of ATG12 to ATG5, and conjugates phosphatidylethanolamine (PE) to LC3 family members. These reactions are essential for autophagosome membrane elongation and closure. ATG3 activity is controlled by nutrient-responsive kinases: mTORC1 suppresses autophagy under fed conditions, whereas AMPK and the ULK1 complex activate it during starvation. Transcriptional regulation by TFEB and FOXO further modulates ATG3 expression. The ATG12?CATG5 conjugate, assembled through ATG3 and ATG10, partners with ATG16L1 to direct LC3 lipidation.
Knockout of ATG3 in 143B cells eliminates autophagosome biogenesis, providing a rigorous autophagy-null background. This system allows precise investigation of autophagy??s roles in osteosarcoma, including its contributions to metabolic stress survival, drug resistance, and metastatic behavior. By comparing wild-type and knockout cells, researchers can elucidate autophagy-dependent signaling crosstalk and identify novel autophagy cargo in a bone tumor context.
These polyclonal knockout cells are validated for standard autophagy assays: western blotting for LC3-II and p62, fluorescence microscopy of LC3 puncta, and flux assays using lysosomal inhibitors. They also support cell viability tests under nutrient deprivation, migration and invasion assays, and in vivo tumor xenograft growth experiments. Applications extend to screening of autophagy modulators, substrate identification, and rescue studies. For comprehensive product support or ordering, contact Ascent Research.
