The ATG3 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population that provides a genetically mixed pool of HeLa cells with disruption of ATG3. This product is intended for investigating the function of ATG3 in autophagy-related pathways without requiring clonal selection. It offers a convenient loss-of-function model for studying ATG3-mediated processes in a widely used human cell line.
HeLa is an immortalized human cervical epithelial cell line derived from a cervical adenocarcinoma and is positive for HPV18. It is a standard host for autophagy research due to its well-characterized signaling networks and robust growth. The epithelial origin and HPV status make it particularly relevant for examining autophagy in the context of oncogenic transformation and host-pathogen interactions.
ATG3 is an E2-like enzyme that catalyzes the conjugation of ATG8/LC3 family proteins to phosphatidylethanolamine, a crucial step in autophagosome membrane expansion. It receives LC3 from the E1 enzyme ATG7 and works with the ATG12-ATG5-ATG16L1 complex to transfer LC3 to lipids. ATG3 activity is regulated upstream by mTOR kinase, AMPK, ULK1 complex, and TFEB transcription factor, and it interacts with ATG7, ATG4B protease, and LC3 isoforms. Key pathway components include LC3B, p62/SQSTM1, Beclin1, and ULK1. Disruption of ATG3 prevents LC3 lipidation, blocking autophagosome formation and cargo engulfment.
In HeLa cells, ATG3 knockout interrupts canonical autophagy flux, making it a valuable tool for dissecting autophagy-dependent phenotypes. Given HeLa??s use in cancer drug resistance studies, the polyclonal knockout cells allow assessment of autophagy??s role in cell survival under chemotherapy or nutrient stress. The HPV18-positive background further enables investigation of viral subversion of autophagic processes, providing insights into infection-associated cancer mechanisms.
Typical experiments include western blotting for LC3 lipidation and immunofluorescence for LC3 puncta formation, often with chloroquine to measure autophagic flux. Cell viability under starvation conditions or drug treatment can reveal ATG3-dependent survival. Co-immunoprecipitation of ATG3 with ATG7 confirms protein interactions, and RT-qPCR profiling of autophagy genes monitors transcriptional changes. The cells are suitable for studying host-pathogen interactions, ER-phagy, and mitophagy, offering a versatile platform for autophagy research. For more information, please contact Ascent Research.
