The EHD4 Knockout K-562 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout population in the K-562 cell line with targeted disruption of the EHD4 gene. This loss-of-function model allows robust investigation of EHD4-mediated endocytic recycling and receptor trafficking without clonal selection, preserving genetic heterogeneity for population-level studies.
The K-562 host cell line originates from a pleural effusion of a 53-year-old female with chronic myelogenous leukemia in blast crisis and bears the BCR-ABL1 fusion. K-562 cells are pluripotent hematopoietic precursors capable of erythroid, granulocytic, and monocytic differentiation, serving as a classic model for erythroleukemia and hematopoietic signaling.
EHD4 encodes a C-terminal EH domain-containing ATPase that localizes to early endosomes and drives receptor recycling. It is activated downstream of EGFR signaling and Rab5/Rab11 GTPases, binding PI(4,5)P2 to induce membrane tubulation. EHD4 dimerization and PI(4,5)P2 binding induce membrane curvature, while interactions with PACSIN2, Rab11-FIP2, cortactin, and the Arp2/3 complex coordinate actin polymerization to drive tubule scission, returning internalized cargos??integrins, transferrin receptor, LDL receptor, and EGFR??to the plasma membrane. This process controls cell adhesion, migration, and signal transduction. EHD4 disruption blocks the Rab5-to-Rab11 conversion, causing endosomal receptor accumulation and attenuated signaling.
In the K-562 leukemia background, EHD4 knockout permits dissection of endocytic recycling in CML pathogenesis. Integrin trafficking is crucial for leukemic cell adhesion and homing to the bone marrow; EHD4 deficiency may impair these processes, potentially affecting metastasis. Crosstalk between EHD4-dependent recycling of growth factor receptors (e.g., EGFR) and BCR-ABL1 signaling could reveal integrative nodes vulnerable in CML. This model is valuable for cancer metastasis, endocytic disorders, and leukemia research.
Researchers can employ this product in diverse assays including Western blotting, RT-qPCR, flow cytometry for surface transferrin/integrin, transferrin recycling kinetics, immunofluorescence for endosomal markers, Transwell migration, and co-immunoprecipitation. These applications support detailed mechanistic studies of endocytic recycling in cancer biology. For further information, please contact Ascent Research.
