The DNM1 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1299 non-small cell lung carcinoma line, in which the dynamin 1 (DNM1) gene has been disrupted. This pool of genetically modified cells provides a powerful loss-of-function model for studying dynamin-dependent membrane trafficking processes without clonal isolation.
The NCI-H1299 cell line was established from a lymph node metastasis of a large cell lung carcinoma and is widely used as a model for lung adenocarcinoma metastasis and drug response. Its well-characterized growth factor signaling pathways, particularly EGFR, make it an ideal host for investigating endocytic regulation of oncogenic signaling.
DNM1 encodes dynamin I, a large GTPase that mediates membrane scission during clathrin-mediated endocytosis. At the neck of invaginating coated pits, dynamin oligomerizes into helical structures and, upon GTP hydrolysis, catalyzes fission to release clathrin-coated vesicles. This process is regulated by upstream kinases such as CDK5, GSK3??, SRC, and PKC, and involves interactions with Amphiphysin, Endophilin, Syndapin, and the AP2 complex. Dynamin-dependent endocytosis directly controls the internalization and subsequent signaling of cell-surface receptors, notably EGFR, thereby influencing downstream AKT and ERK activation. Additionally, dynamin participates in mitochondrial fission and mTOR signaling networks.
In NCI-H1299 lung cancer cells, dynamin I governs surface levels of EGFR and other receptor tyrosine kinases, positioning it as a critical node in pro-proliferative and migratory signaling. Loss of DNM1 disrupts EGFR internalization and degradation, leading to altered phospho-signaling dynamics that can impact cell migration and invasion. This knockout model thus enables precise dissection of endocytic contributions to cancer cell behavior, including receptor trafficking and downstream pathway activation.
Typical applications include functional analysis of clathrin-mediated endocytosis, investigation of EGFR trafficking and degradation, and assessment of dynamin??s role in cell migration and invasion. Researchers can employ transferrin uptake assays to quantify bulk endocytosis or immunofluorescence staining for clathrin-coated pit markers. EGFR degradation kinetics can be monitored by Western blotting, while phospho-AKT and phospho-ERK analysis reveals signaling consequences. Live-cell imaging of endocytosis, co-immunoprecipitation of dynamin complexes, and drug target assessment for endocytosis inhibitors are also feasible. For detailed technical inquiries and to explore how this polyclonal knockout model can support your research, please contact Ascent Research.
