The ATG7 Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human osteosarcoma cell line 143B. This loss-of-function model disrupts the ATG7 gene, which encodes the core autophagy E1 enzyme, and is supplied as a polyclonal population to minimize clonal artifacts while ensuring robust target gene disruption. It provides a genetically stable system for dissecting autophagy-dependent mechanisms in cancer biology.
The host 143B line is a well-characterized human osteosarcoma model, originating as a TK-negative derivative of HOS cells. It is widely employed in metastasis and tumorigenesis research due to its aggressive invasive properties and faithful recapitulation of bone cancer phenotypes. This background makes the ATG7 knockout cells particularly relevant for studying autophagy??s contributions to osteosarcoma progression and therapeutic response. The cells are extensively used for in vivo xenograft studies and in vitro motility assays.
ATG7 acts as the essential E1 enzyme for two ubiquitin-like conjugation systems vital for autophagosome formation. It activates ATG12 and transfers it to ATG10, enabling conjugation to ATG5, which assembles into the ATG12?CATG5?CATG16L1 complex. This complex functions as an E3 ligase for LC3 lipidation. In parallel, ATG7 primes LC3 family members and hands them to ATG3 for conjugation to phosphatidylethanolamine, generating the membrane-bound LC3-II form that drives autophagosome elongation and closure. ATG7 activity is governed by mTORC1 inhibition and AMPK activation, downstream of transcription factors like TFEB, FOXO3, p53, and E2F1. It interacts with ATG3, ATG10, ATG12, the ATG8/LC3 family, and the autophagy receptor p62/SQSTM1. This network integrates with the ULK1?CBeclin1?CVPS34 initiation machinery, placing ATG7 at a central hub for macroautophagy, selective autophagy, and mitophagy.
In the context of human osteosarcoma, the ATG7 knockout model is a powerful tool for examining how autophagy supports bone cancer cell survival, metastasis, and drug resistance. Osteosarcoma cells often rely on autophagic degradation to cope with nutrient deprivation and chemotherapeutic stress; ablation of ATG7 effectively blocks this catabolic process, enabling researchers to dissect autophagic dependencies in tumorigenesis. This model facilitates investigation of crosstalk between autophagy and signaling pathways frequently dysregulated in osteosarcoma, such as mTOR and AMPK, and aids in the identification of therapeutic vulnerabilities.
Key research applications include autophagy flux analysis using tandem fluorescent LC3 reporters by flow cytometry, assessing LC3-II and p62 levels by western blotting, and ultrastructural examination of autophagic structures by electron microscopy. Additional uses encompass cell viability, apoptosis, migration, and invasion assays, as well as drug sensitivity profiling with autophagy modulators. This product supports gene therapy validation and chemoresistance studies in a clinically relevant bone cancer cell background. For further details, please contact Ascent Research.
