Diverse Role of Atg Proteins in Autophagy and Cellular Homeostasis

Autophagy plays a critical role in cellular homeostasis by mediating the degradation and recycling of cellular components. The objective of this study was to explore the specific roles of Atg proteins in the autophagy pathway. Utilizing CRISPR-Cas9 gene editing, we generated Atg-deficient cell lines to elucidate the role of these proteins. Through live-cell imaging and Western blot analysis, we identified significant disruptions in autophagosomal membrane formation and cargo degradation in Atg5 and Atg7 knockout models. Quantitative assays revealed a 65% reduction in LC3-II conversion and a 78% decrease in p62 degradation in Atg5-deficient cells compared to controls. Furthermore, electron microscopy confirmed the accumulation of undegraded autophagic vesicles, underscoring impaired autophagic flux. These findings illustrate the indispensable role of Atg proteins in maintaining autophagic function and cellular health. We conclude that targeted manipulation of these proteins could lead to novel therapeutic strategies for diseases associated with autophagy dysregulation, such as neurodegenerative disorders and cancer.