The EHBP1 Knockout NCI-H1299 Polyclonal Cells product provides a pooled population of NCI-H1299 cells subjected to CRISPR/Cas9-mediated disruption of the EHBP1 gene. This polyclonal knockout cell population, derived from the human non-small cell lung cancer line NCI-H1299, serves as a loss-of-function model for investigating EHBP1-dependent endocytic recycling, cytoskeletal dynamics, and cell migration. Unlike clonal isolates, the polyclonal composition preserves population-level heterogeneity while eliminating wild-type EHBP1 expression, enabling studies where mosaic gene disruption is acceptable or preferred.
The parental NCI-H1299 line was originally established from a lymph node metastasis of a lung adenocarcinoma and is widely utilized as a model of metastatic non-small cell lung cancer. These cells retain key features of invasive adenocarcinoma, including active endocytic trafficking and actin-based motility, making them a physiologically relevant host for examining EHBP1 function in the context of cancer cell dissemination.
EHBP1 encodes a scaffold protein that physically links Rab8a-decorated recycling endosomes to the cortical actin cytoskeleton. This interaction is mediated through its EH-domain-binding partners EPS15 and EHD1, which together couple endocytic trafficking to actin polymerization. Mechanistically, EHBP1 functions downstream of activated Rab8a and is subject to regulation by insulin/PI3K/Akt signaling and growth factors such as EGF and insulin. Once recruited to endosomal membranes, EHBP1 promotes nucleation of actin filaments via the N-WASP?CArp2/3 complex, thereby driving both GLUT4 vesicle translocation to the plasma membrane in insulin-responsive contexts and the formation of actin-based protrusions required for cell migration. Additional interacting factors include Bin1, which further modulates membrane-actin coupling.
In the NCI-H1299 lung adenocarcinoma background, EHBP1 acts as a node connecting endosomal recycling to the motile apparatus of the cell. Its disruption impairs the efficient recycling of cargoes such as GLUT4 and likely other surface receptors, attenuating cellular responses to growth factors and metabolic cues that support metastasis. Consequently, the knockout model aids in dissecting how dysregulated endocytic trafficking contributes to the invasive phenotype and may reveal vulnerabilities that can be exploited therapeutically.
This polyclonal knockout product is suited for a variety of experimental workflows aimed at elucidating EHBP1 biology in cancer. Researchers can employ Transwell migration and invasion assays to quantify metastatic potential, immunofluorescence staining to assess GLUT4 subcellular localization or actin organization, and transferrin uptake assays to measure general endocytic recycling. Co-immunoprecipitation and Western blotting enable verification of disrupted EHBP1?CEPS15?CEHD1 complex formation and downstream signaling changes such as phospho-Akt levels following insulin stimulation. The cells also support functional screens for protein interactions and drug sensitivity studies. For pricing, availability, or technical consultation, please contact Ascent Research.
