The EHD3 Knockout NCI-H1299 Polyclonal Cells product consists of a polyclonal knockout cell population derived from the NCI-H1299 non-small cell lung carcinoma cell line, engineered via CRISPR/Cas9-mediated disruption of the EHD3 gene. This loss-of-function model eliminates EHD3 expression across a heterogeneous cell pool, enabling the study of EHD3-dependent cellular processes without clonal artifacts.
The NCI-H1299 host cell line is a widely used model of lung adenocarcinoma, established from a lymph node metastasis. It harbors a homozygous deletion of TP53, rendering it deficient in p53 function, and exhibits an epithelial morphology. These characteristics make NCI-H1299 particularly suitable for investigating tumor cell biology, including proliferation, migration, and responses to therapeutic agents.
EHD3 functions as an endocytic recycling protein that regulates the intracellular trafficking of membrane receptors, notably epidermal growth factor receptor (EGFR) and integrin ??1. It interacts with EHD1, SNX9, the AP-2 complex, Rab11-FIP3, and actin nucleation factors such as N-WASP. Upstream, EHD3 is activated by receptor engagement (e.g., EGF, PDGF), and it then controls the recycling of EGFR and integrins, thereby modulating downstream signaling through mTORC1, MAPK (ERK), and Akt pathways. By facilitating receptor return to the plasma membrane, EHD3 sustains signaling intensity, influences actin cytoskeleton reorganization, and affects cell adhesion and motility.
In the NCI-H1299 background, knockout of EHD3 disrupts the balance of endocytic recycling, potentially attenuating EGFR and integrin-mediated signaling. Given the context-dependent roles of EHD3??acting as a tumor suppressor in some settings and an oncogenic factor in others??this polyclonal knockout model provides a valuable tool for dissecting its function in non-small cell lung cancer. Alterations in receptor trafficking may impact cell proliferation, migration, and sensitivity to targeted therapies, including EGFR inhibitors and mTOR antagonists.
This product is ideally suited for a broad range of research applications, including endocytic trafficking studies, cancer cell migration and invasion assays, drug resistance screening, and signaling pathway analysis. Representative techniques include Western blotting to confirm EHD3 depletion, quantitative RT-PCR for transcript validation, flow cytometry-based receptor recycling assays, wound healing and transwell invasion experiments, and phospho-protein arrays to monitor downstream effectors like phospho-S6K, phospho-ERK, and phospho-Akt. Additionally, confocal microscopy can visualize altered localization of endocytic markers such as Rab11. For further information, please contact Ascent Research.
