The EHD4 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population designed for loss-of-function studies of the EHD4 gene. Derived from the HAP1 near-haploid cell line via gene disruption, these cells offer a robust model system to investigate endosomal recycling and receptor trafficking. The polyclonal format provides a heterogeneous knockout background, avoiding clonal selection bias and more closely reflecting physiological gene disruption.
HAP1 cells are a near-haploid human cell line originating from a male patient with chronic myeloid leukemia. Their unique haploid karyotype simplifies genetic analyses by eliminating the confounding effects of a second allele, ensuring unequivocal phenotypic linkage to the targeted gene. The fibroblast-like morphology and retention of key endocytic pathways make HAP1 an ideal host for dissecting membrane trafficking mechanisms. This cell line is widely recognized as a gold standard for CRISPR-based knockout studies due to its genetic tractability and reproducible culture characteristics.
EHD4 encodes an ATPase that regulates endosomal recycling by promoting membrane tubulation and fission. It functions downstream of receptor activation and phosphoinositide signaling and interacts with EHD1, EHD2, Rab GTPases, and syndapin/PACSIN proteins to control the recycling of internalized receptors such as transferrin receptor and integrins. By coupling ATP hydrolysis to membrane remodeling, EHD4 facilitates the return of these receptors to the plasma membrane, thereby modulating cell adhesion, migration, and signaling. Loss of EHD4 disrupts this recycling, leading to altered surface receptor levels and impaired cellular responses.
In the near-haploid HAP1 context, EHD4 disruption yields a clean loss-of-function model that avoids the complexity of diploid compensation. The polyclonal knockout pool captures the spectrum of gene-editing events, providing a population-based tool for interrogating endosomal trafficking. This configuration is especially valuable for high-content screening and assays that benefit from population-level readouts, such as transferrin recycling kinetics and integrin trafficking. The simplified genetic background ensures that observed phenotypes are directly attributable to EHD4 deficiency, facilitating unambiguous interpretation.
Research applications include endosomal trafficking studies, receptor recycling assays, cell adhesion and migration analyses, and neurological disease modeling. Representative assays with this product include transferrin uptake and recycling by flow cytometry, integrin recycling using antibody-feeding protocols, immunofluorescence for endosomal markers, and co-immunoprecipitation to assess protein interactions. Cell migration and invasion assays further define the role of EHD4 in cancer biology. For additional information, please contact Ascent Research.
