The ATG9A Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the autophagy-related gene ATG9A has been disrupted. This heterogeneous population of HAP1 cells carries diverse ATG9A mutations introduced via non-homologous end joining following Cas9 cleavage, providing a loss-of-function model suitable for studying ATG9A-dependent processes without the biases of clonal selection. The polyclonal format ensures a representative knockout background for robust downstream functional analyses.
The HAP1 host cell line is a near-haploid human male cell line derived from the KBM-7 chronic myeloid leukemia cell line and adapted to suspension growth. Its near-haploid karyotype simplifies genetic manipulation and enhances knockout generation efficiency, while the suspension growth mode supports scalable culture for high-throughput and biochemical applications. HAP1 cells retain core signaling pathways, including fully functional autophagic machinery, making them an optimal platform for dissecting the roles of autophagy-related genes such as ATG9A.
ATG9A encodes a multispanning transmembrane protein that dynamically cycles between the trans-Golgi network, endosomes, and the phagophore assembly site, where it supplies membrane essential for autophagosome biogenesis. Its activity is stimulated by nutrient deprivation and mTORC1 inhibition, mediated by AMPK activation and the ULK1 complex (ULK1, ATG13, FIP200, ATG101). ATG9A interacts with the PI3K-III complex (Beclin-1, VPS34, ATG14) and with ATG2A, WIPI1, and WIPI2 to coordinate membrane expansion. Downstream, ATG9A is indispensable for LC3 (MAP1LC3B) lipidation, p62/SQSTM1 degradation, and autophagosome maturation, ultimately regulating autophagic flux.
In HAP1 cells, ATG9A knockout creates a clean genetic background to investigate autophagy-dependent cancer cell survival. Derived from chronic myeloid leukemia, HAP1 cells model hematological malignancy, allowing researchers to examine how disruption of membrane delivery at the phagophore affects proliferation, stress responses, and chemotherapeutic sensitivity. This polyclonal knockout population is particularly valuable for genetic complementation assays to validate ATG9A function and for studying autophagy-related drug resistance mechanisms in a near-haploid context.
This product supports diverse experimental applications, including autophagy pathway analysis via western blotting for LC3-II and p62/SQSTM1, immunofluorescence staining of LC3 puncta, and autophagic flux measurements using bafilomycin A1. It is well-suited for high-throughput screening of autophagy modulators, functional genomic screens leveraging the near-haploid background, and drug sensitivity profiling. For further details, please contact Ascent Research.
