The EHD1 Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout population, providing a loss-of-function model for EHD1 in HeLa cervical adenocarcinoma cells. This heterogeneous pool of cells carries targeted disruptions of the EHD1 locus, enabling robust functional studies free from clonal selection artifacts. The polyclonal format preserves parental line diversity, ensuring reproducibility in downstream assays.
HeLa cells are an immortalized epithelial cell line derived from an HPV18-positive cervical adenocarcinoma, widely used in cancer biology and signaling research. They retain HPV oncogene expression, which inactivates p53 and Rb, and exhibit robust growth and genetic tractability ideal for CRISPR editing. Their epithelial origin and invasive properties make them a valuable model for studying membrane trafficking in cancer progression.
EHD1 encodes an ATPase that functions as a key regulator of endocytic receptor recycling. EHD1 is recruited to phosphatidylinositol-4-phosphate (PI(4)P)-enriched membranes of recycling endosomes, where it oligomerizes and uses ATP hydrolysis to generate mechanical force driving membrane fission. This process facilitates the return of internalized cargo, including the transferrin receptor (TFRC) and EGFR, to the plasma membrane. EHD1 operates in a complex network with Rab4 and Rab5, which regulate its recruitment, and with ARF6, syndapin II, amphiphysin, and EHBP1, which cooperate in membrane remodeling. Through these interactions, EHD1 also controls recycling of integrin receptors, thus influencing cell adhesion and migration.
In the HeLa cell context, disruption of EHD1 is expected to impair endosomal recycling, leading to altered surface expression of EGFR and integrins, which are critical for proliferation and migration. The HPV18-positive cervical adenocarcinoma background may reveal unique dependencies on receptor trafficking pathways. Consequently, this polyclonal knockout model provides a physiologically relevant platform to dissect EHD1??s contributions to tumor cell behavior and its potential as a therapeutic target.
These EHD1 knockout cells are optimized for a broad range of functional assays, including transferrin recycling kinetics to directly measure endosomal trafficking, EGFR degradation and signaling dynamics by western blotting and flow cytometry, and migration and invasion assays to assess metastatic potential. Co-immunoprecipitation studies can map protein interactions within the EHD1 complex, and immunofluorescence can visualize receptor mistrafficking. The cells are also suitable for screening small-molecule inhibitors targeting EHD1-dependent pathways and for high-content phenotypic screens. For inquiries, contact Ascent Research.
