The EHD3 Knockout AGS Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population derived from the AGS human gastric adenocarcinoma cell line, engineered to disrupt the EHD3 gene. This polyclonal population consists of a heterogeneous mix of edited cells with targeted gene disruption, serving as a loss-of-function model to study EHD3-dependent processes. The knockout cells enable investigation of endocytic recycling, receptor trafficking, and membrane homeostasis without the confounding effects of single-cell clonal variation, thus representing a physiologically relevant genetic knockout system for broad-screening and functional analysis.
AGS cells are an adherent epithelial cell line originally isolated from a human gastric adenocarcinoma. They retain characteristics of gastric epithelium, including the expression of cell surface receptors such as EGFR and integrins, making them a well-established model for gastric cancer biology. The AGS background provides a relevant context for probing the role of EHD3 in the maintenance of epithelial polarity, receptor-mediated signaling, and oncogenic transformation associated with gastric malignancies. The polyclonal knockout configuration in this cell line offers a stable system for investigating genetic dependencies in a tumor cell background.
EHD3 (EH domain-containing protein 3) functions as a key regulator of endocytic recycling, mediating the return of internalized cargo from endosomes back to the plasma membrane. Mechanistically, EHD3 interacts with Arf6 and Rab11, members of the small GTPase families that coordinate membrane trafficking and actin remodeling. Upstream growth factors and EGFR signaling activate EHD3, localizing it to recycling endosomes where it facilitates the retrieval of activated receptors such as EGFR and integrins, thereby controlling the intensity and duration of downstream signaling cascades. Loss of EHD3 disrupts this recycling pathway, leading to aberrant receptor accumulation and altered actin cytoskeleton dynamics, which can impact cell migration and proliferation.
In gastric adenocarcinoma, receptor trafficking pathways are commonly dysregulated, fueling sustained proliferative signaling, metastatic dissemination, and therapeutic resistance. EHD3 disruption in AGS cells impairs the coordinated recycling of EGFR and integrins, resulting in altered downstream signaling through pathways such as the mitogen-activated protein kinase (MAPK) cascade and the actin regulatory network. This knockout model therefore allows researchers to dissect how endosomal sorting contributes to gastric cancer pathogenesis and to evaluate the role of EHD3 in maintaining normal epithelial homeostasis versus promoting malignant phenotypes.
This EHD3 knockout product is suitable for a range of advanced research applications, including cancer cell biology, receptor trafficking studies, drug resistance mechanisms, and migration and invasion assays. Researchers can utilize western blotting to assess protein expression changes, immunofluorescence microscopy to examine subcellular localization of EGFR and actin, receptor internalization and recycling assays with phospho-EGFR analysis, and functional migration assays to evaluate cellular motility. Additional applications include RT-qPCR profiling of downstream targets and signaling intermediates. For further information or technical support, contact Ascent Research.
