The AMBRA1 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from A-549 lung adenocarcinoma cells, carrying targeted disruption of the AMBRA1 gene. This loss-of-function model enables ablation of AMBRA1 protein expression and provides a versatile tool for studying autophagy regulation and cell cycle control in a human cancer background. The polyclonal format captures diverse editing events while maintaining robust experimental reproducibility.
A-549 cells originated from a 58-year-old male lung adenocarcinoma and exhibit type II alveolar epithelial properties, including surfactant production and xenobiotic metabolism. These cells harbor an oncogenic KRAS G12S mutation, making them a well-established model for KRAS-driven lung cancer research. Their epithelial origin and genetic stability render them ideal for mechanistic studies of tumor biology and drug responses.
AMBRA1 functions as a scaffold protein that stabilizes the Beclin1-Vps34 PI3K complex, facilitating autophagosome nucleation and LC3 lipidation, and acts as a substrate receptor for the DDB1-CUL4 E3 ligase to target cyclin D for degradation. Upstream regulators include mTORC1, ULK1, p53, and AMPK, while downstream effects involve p62 degradation, mitochondrial clearance, and apoptosis modulation via BCL2 binding. AMBRA1 thus coordinates autophagy with cell cycle progression.
In A-549 cells, AMBRA1 knockout impairs autophagy flux and disrupts cyclin D proteolysis, providing a system to examine how autophagy intersects with proliferation under KRAS-driven conditions. This model is relevant for investigating tumor suppressive mechanisms, as AMBRA1 loss is linked to lung adenocarcinoma, melanoma, and breast cancer. It also permits analysis of chemoresistance and stress adaptation pathways in alveolar epithelial cancer cells.
This polyclonal knockout product is suited for assays such as LC3 turnover with bafilomycin A1, p62 immunofluorescence, GFP-LC3 puncta counting, flow cytometric cell cycle analysis, and cyclin D degradation assays. It supports drug screening for autophagy modulators and studies of cisplatin sensitivity and apoptosis. For technical inquiries or ordering, please contact Ascent Research.
