The ATG5 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population designed to abolish expression of the autophagy core protein ATG5 in the near-haploid HAP1 cell line. This loss-of-function pool enables bulk interrogation of autophagy without clonal expansion, offering a versatile reagent for genetic and pharmacological studies.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) model. Its haploid genome simplifies functional genomics by eliminating diploid gene redundancy, making it a favored platform for knockout screens and pathway dissection. In cancer research, HAP1 provides a relevant background for studying tumor-related processes such as autophagy.
ATG5 is a central autophagy effector. Upon activation by ULK1, AMPK, or mTORC1 signaling, ATG5 is conjugated to ATG12 via the E1 enzyme ATG7 and E2 enzyme ATG10. The ATG5?CATG12 complex dimerizes with ATG16L1 to form an E3-like ligase that lipidates LC3/GABARAP proteins, driving autophagosome expansion and maturation. This process links nutrient deprivation, hypoxia, and ER stress to autophagic degradation, positioning ATG5 as a critical integrator of metabolic and stress signals.
In the HAP1 context, ATG5 knockout creates a robust model to analyze autophagy-dependent phenotypes in a cancer cell line. Its near-haploid state ensures clear genotype-phenotype relationships, allowing direct attribution of impaired LC3 puncta formation, reduced autophagic flux, or increased susceptibility to nutrient stress to ATG5 deficiency. This is particularly valuable for dissecting autophagy??s dual role in tumor suppression and oncogenesis.
Typical applications include western blot analysis of LC3-II/LC3-I ratios to monitor flux, fluorescence microscopy of GFP-LC3 puncta, co-immunoprecipitation to detect ATG5?CATG12 complex disruption, and cell viability assays under starvation. The polyclonal format is well suited for high-throughput screening of autophagy modulators and for investigating host?Cpathogen interactions where autophagic machinery is co-opted. For further details, please contact Ascent Research.
