The EHBP1 Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the EHBP1 gene in a human hematopoietic suspension background. This product provides a loss-of-function model to study the roles of the multidomain adaptor EHBP1 in endocytic recycling and actin cytoskeleton reorganization. The polyclonal pool preserves cellular heterogeneity and allows robust functional screening without selection-induced clonal bias. As a ready-to-use polyclonal knockout resource, it eliminates the need for researchers to perform in-house gene editing.
The parental K-562 cell line is a well?characterized suspension line derived from a patient with Philadelphia chromosome?positive chronic myeloid leukemia (CML) in blast crisis. K-562 cells display hallmarks of multipotent hematopoietic progenitors, with capacity to differentiate along erythroid, megakaryocytic, and monocytic lineages under appropriate stimuli. Their BCR?ABL1?driven oncogenic signalling makes them a standard model for CML biology and drug development, particularly for investigating tyrosine kinase inhibitor resistance. The cells grow in suspension, facilitating high?throughput screening, flow cytometry?based assays, and biochemical fractionation without trypsinization.
EHBP1 is a scaffold protein that links EHD?family ATPases to the actin machinery at recycling endosomes. Activated downstream of Cdc42 GTPase, it interacts with EHD1, EHD2, cortactin, and F?actin to coordinate membrane tubulation and actin assembly. The protein bridges EHD1?positive endosomes to the N?WASP/Arp2/3 complex, promoting localized actin nucleation that supports Rab8a? and Rab11?dependent cargo sorting and recycling of receptors like EGFR and GLUT4. Thus, EHBP1 integrates Cdc42 signalling with endocytic traffic, shaping plasma membrane composition and receptor dynamics.
In the K-562 myeloid leukemia background, EHBP1 disruption provides a relevant setting to dissect crosstalk between oncogenic BCR?ABL1 signalling and the actin?dependent endocytic machinery. EHBP1?deficient K-562 cells enable investigation of how altered recycling endosome dynamics modify surface abundance of adhesion molecules and receptor tyrosine kinases, potentially affecting cell?substrate interactions and chemotactic migration. Given that EHBP1 interacts with cortactin??a BCR?ABL1 substrate??this model can uncover links between leukemogenic kinase activity and cytoskeletal remodelling. The polyclonal knockout population preserves pathway heterogeneity, making it suitable for drug sensitivity profiling and resistance mechanism studies.
Researchers can use EHBP1 Knockout K-562 Polyclonal Cells for transferrin and dextran uptake assays to quantify endocytic capacity, immunofluorescence staining to visualize EHD1 and cortactin distribution, and flow cytometry to measure surface receptor recycling. Boyden chamber assays provide readouts for migration, while actin polymerization assays directly assess N?WASP/Arp2/3 activity. Additional applications include western blotting and RT?qPCR for pathway validation. For technical support and collaborative studies, please contact Ascent Research.
