The ATG5 Knockout A2780 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A2780 human ovarian carcinoma cell line, engineered to disrupt the ATG5 gene. This product offers a heterogeneous mixture of edited cells, retaining the native genetic diversity of the parental line while abolishing ATG5 expression. Supplied as a ready-to-use polyclonal stock, it enables straightforward functional dissection of ATG5-dependent pathways without the need for single-cell cloning, making it ideal for pooled autophagy and cancer studies.
The A2780 cell line originates from an untreated patient with ovarian carcinoma and serves as a widely employed model for high-grade serous ovarian adenocarcinoma. These epithelial cells display characteristics consistent with ovarian cancer biology, including sensitivity to platinum- and taxane-based chemotherapeutics. Consequently, A2780 is a standard choice for drug response profiling, signal transduction analyses, and metastasis research, providing a clinically relevant background for studying autophagy in ovarian malignancy.
ATG5 is an indispensable autophagy protein that forms a conjugate with ATG12 via the E1-like enzyme ATG7 and the E2-like enzyme ATG10. The ATG12?CATG5 conjugate associates with ATG16L1 to generate a multimeric complex that functions as an E3-like enzyme, driving the lipidation of LC3 and GABARAP family members during phagophore elongation. This lipidation event is essential for autophagosome membrane expansion and cargo sequestration. Beyond autophagy, ATG5 participates in apoptosis: calpain-mediated cleavage yields an N-terminal fragment that translocates to mitochondria, promoting cytochrome c release. ATG5 also intersects with innate immune signaling, modulating type I interferon responses. The ATG5-dependent system is regulated by upstream kinases, including mTORC1, which suppresses autophagy under nutrient-rich conditions, and AMPK and the ULK1 complex, which activate autophagy during starvation. The Beclin1?CVPS34 complex further contributes to phagophore nucleation. Key interacting partners include ATG16L1, TECPR1, calpains, BCL-2 family members, and TRAF6.
In ovarian carcinoma, autophagy has been implicated in both tumor suppression and promotion, particularly in the development of chemoresistance. The A2780 knockout model enables precise interrogation of how ATG5-mediated autophagy influences cellular responses to cisplatin and paclitaxel, two frontline therapies for ovarian cancer. Disruption of ATG5 also allows researchers to dissect the crosstalk between autophagic and apoptotic pathways, as calpain-mediated cleavage of ATG5 directly links these processes. Moreover, this model is valuable for examining autophagy??s role in tumor cell invasion, metastasis, and immune evasion, which are major drivers of poor prognosis in high-grade serous ovarian adenocarcinoma.
These polyclonal knockout cells support a broad range of applications, including autophagy flux analysis using LC3-II turnover and p62 degradation assays, investigation of apoptosis through Annexin V/PI flow cytometry, and drug sensitivity testing with platinum- or taxane-based compounds. Co-immunoprecipitation of ATG12?CATG5 complexes confirms the loss of ATG5 expression, while immunofluorescence for LC3 puncta provides visual readouts of autophagosome formation. Additional uses include transwell migration/invasion assays and RT-qPCR analysis of autophagy-related genes. For further technical information, batch-specific validation data, or custom requests, please contact Ascent Research.
