EHD3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for studying endocytic recycling and related cellular processes. This product provides a pool of HAP1 cells with CRISPR/Cas9-mediated disruptions in the EHD3 gene, enabling loss-of-function analysis without clonal selection. The polyclonal format avoids clonal artifacts and maintains population diversity, suitable for pooled screening and pathway investigations. Researchers can use this model to explore EHD3 roles in membrane trafficking, receptor recycling, and actin dynamics.
The HAP1 (human near-haploid chronic myeloid leukemia) cell line is a genetic model derived from KBM-7, haploid for most chromosomes except a duplicated region of chromosome 8. This near-haploid state facilitates straightforward gene targeting, as single-copy gene disruption generates knockout phenotypes. HAP1 cells retain key endocytic and signaling pathways, making them ideal for investigating genes involved in receptor trafficking, including EHD3. These cells are widely used in haploid genetic screens and endocytosis studies.
EHD3 (EH domain-containing protein 3) is an ATPase that functions downstream of Rab5 and Rab11 in endocytic recycling, mediating membrane tubulation and fission to return internalized receptors such as transferrin receptor and EGFR to the plasma membrane. It interacts with syndapin, amphiphysin, Rab11-FIP2, EHD1, and EHD4, and coordinates with Arf6 and actin polymerization machinery to link vesicular trafficking to cytoskeletal reorganization, thereby regulating cell migration, adhesion, and signal attenuation. Dysregulation of EHD3 is associated with cancer metastasis and atrial fibrillation.
In HAP1 cells, EHD3 knockout disrupts the recycling of surface receptors, providing a clear phenotypic readout. The haploid nature of HAP1 ensures that CRISPR/Cas9-mediated editing leads to a penetrant loss-of-function phenotype at the population level, without the need for homozygous editing. This cell model is particularly valuable for dissecting the contributions of EHD3 to integrin trafficking, EGFR signaling dynamics, and actin-mediated membrane remodeling. Because HAP1 cells express a near-normal endocytic machinery, the EHD3 knockout polyclonal population allows direct observation of defects in transferrin recycling, receptor surface expression, and cell migration.
Typical applications of EHD3 Knockout HAP1 Polyclonal Cells include western blotting to confirm EHD3 depletion, transferrin recycling assays to quantify endocytic flux, and flow cytometry to measure surface transferrin receptor or EGFR levels. These cells are suited for wound healing or transwell migration assays to evaluate EHD3’s role in cell motility, and for haploid CRISPR screens to identify genetic interactions in the endocytic pathway. In cancer research, this model can study drug resistance mechanisms mediated by altered receptor trafficking. For further information, please contact Ascent Research.
