The ATG7 Knockout Ca Ski Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the autophagy-related 7 (ATG7) gene in the human cervical squamous cell carcinoma cell line Ca Ski. This genetically heterogeneous pool of ATG7-null cells enables rigorous loss-of-function studies of autophagy-dependent processes, circumventing artifacts associated with clonal selection and providing a more physiologically relevant model of gene disruption.
The Ca Ski host cell line originates from a cervical lymph node metastasis of squamous cell carcinoma and harbors approximately 600 integrated copies of the human papillomavirus type 16 (HPV-16) genome. Persistent expression of the viral E6 and E7 oncoproteins leads to destabilization of the tumor suppressors p53 and Rb, respectively, driving unchecked proliferation and genomic instability. This well-established line is extensively used to model HPV-associated carcinogenesis, investigate oncogene-driven signaling, and assess responses to conventional and targeted therapies.
ATG7 encodes an E1-like activating enzyme that is central to canonical autophagy. It catalyzes the ATP-dependent adenylation and activation of ATG12 and the ATG8 family??including MAP1LC3A/B/C (LC3) and GABARAP subfamilies??and transfers them to the E2 conjugating enzymes ATG10 and ATG3, respectively. This activity drives the covalent conjugation of ATG12 to ATG5, which forms a complex with ATG16L1, and the lipidation of LC3 to phosphatidylethanolamine (LC3-II), a prerequisite for autophagosome membrane expansion and closure. Key upstream regulators include mTORC1, which suppresses autophagy via phosphorylation of ULK1, and AMPK, which promotes autophagy by activating ULK1 and inhibiting mTORC1. The transcription factors FOXO1/3, TFEB, and ATF4 further control ATG7 expression. Additionally, ATG7 interacts with the cargo receptor p62/SQSTM1, facilitating the selective degradation of ubiquitinated proteins.
In the Ca Ski cellular context, deletion of ATG7 abolishes autophagic flux, leading to accumulation of autophagic substrates such as p62 and potentially impairing the autophagy-mediated degradation of HPV E6 and E7 oncoproteins, which are known targets of selective autophagy. This loss may disrupt cellular metabolism, elevate oxidative stress, and compromise proteostasis, thereby altering the sensitivity of HPV-positive cervical carcinoma cells to chemotherapeutic agents, such as cisplatin, and to inhibitors of the PI3K/AKT/mTOR pathway. Consequently, the ATG7 knockout polyclonal pool serves as a powerful tool for dissecting autophagy-dependent tumor cell survival, HPV oncoprotein turnover, and therapeutic resistance mechanisms.
Researchers can leverage this ATG7-knockout Ca Ski pool to explore autophagy mechanisms, HPV oncoprotein degradation, metabolic reprogramming, drug resistance, and tumorigenesis. A broad range of established assays is compatible, including Western blotting for LC3 conversion and p62/SQSTM1 clearance, measurement of autophagic flux using lysosomal inhibitors (bafilomycin A1, chloroquine), fluorescence imaging of GFP-LC3 puncta, cell viability and apoptosis assays under nutrient starvation or drug treatment, co-immunoprecipitation of ATG5-ATG12 complexes, Seahorse-based metabolic profiling, and in vivo tumor xenograft growth analysis. For additional information or custom inquiries, please contact Ascent Research.
