The ATG10 Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for studying autophagy-related processes. This product provides a genetically disrupted ATG10 gene in the HeLa cell background, enabling loss-of-function studies of the E2-like enzyme essential for the ATG12-ATG5 conjugation cascade.
HeLa cells are an immortalized epithelial model derived from an HPV18-positive cervical adenocarcinoma. Their robust growth, well-characterized signaling pathways, and extensive literature support make them a standard platform for cancer biology and molecular cell biology research. The cervical adenocarcinoma origin positions this line for studies in oncogenic transformation and tumor cell survival mechanisms.
ATG10 serves as the E2-like enzyme in the ATG12 conjugation system, collaborating with the E1-like enzyme ATG7 to covalently attach ATG12 to ATG5. The resulting ATG12-ATG5 conjugate further associates with ATG16L1 to form a multimeric complex that functions as an E3-like enzyme for the lipidation of LC3-I to LC3-II, a critical step in autophagosome membrane expansion. Upstream, ATG10 expression and autophagy initiation are tightly regulated by nutrient-sensing pathways, including mTORC1 and AMPK, which control the ULK1 complex. Additionally, the transcription factor TFEB governs the expression of ATG10 and other autophagy-related genes. Disruption of ATG10 thus impedes the ATG12-ATG5 conjugation step, blocking LC3 lipidation and autophagosome formation.
In the HPV18-positive cervical adenocarcinoma context, autophagy modulation influences tumor cell survival, drug resistance, and metabolic adaptation. Ablation of ATG10 in these polyclonal knockout HeLa cells provides a well-defined model to dissect autophagy-dependent phenotypes in cancer. This tool enables precise interrogation of how the ATG12?CATG5 conjugation pathway contributes to proliferation, apoptosis avoidance, and response to chemotherapeutics, thereby offering insights relevant to cervical cancer and other malignancies where autophagy is deregulated.
These polyclonal knockout cells are suitable for a variety of applications, including autophagy flux analysis using western blot detection of LC3-II turnover in the presence of lysosomal inhibitors such as chloroquine, immunofluorescence imaging of LC3 puncta, and cell viability assays under stress conditions. They also serve as a validated CRISPR knockout model for drug discovery campaigns targeting the autophagy pathway and for studying protein degradation dynamics. This product is an essential resource for investigating autophagy-mediated survival mechanisms. For additional technical information and support, please contact Ascent Research.
