The CCDC22 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells, designed to disrupt CCDC22 gene expression. This heterogeneous pool provides a robust loss-of-function model for studying the role of the COMMD/CCDC22/CCDC93 (CCC) complex in endosomal cargo sorting and downstream signaling events. The polyclonal format preserves biological variability and is ideal for population-based functional assays.
The parental HeLa cell line is an immortalized human cervical epithelial carcinoma line harboring HPV18-positive adenocarcinoma. HeLa cells are widely utilized in cancer biology and cell signaling research due to their robust growth, genetic tractability, and well-characterized endosomal trafficking machinery. Their epithelial origin makes them particularly suitable for examining membrane receptor recycling, copper homeostasis, and the CCC complex’s role in these processes.
CCDC22 is a core subunit of the CCC complex, which scaffolds the WASH complex onto endosomal membranes. This interaction promotes WASH-dependent actin polymerization, facilitating the recycling of cargo such as the copper transporters ATP7A and ATP7B and the Notch1 receptor. Upstream regulators including Rab5 and endosomal cues coordinate this activity, while the CCC complex interacts with COMMD proteins (COMMD1-10), CCDC93, and WASH components (WASHC1, WASHC4, WASHC5, FAM21, Strumpellin, SWIP). Additionally, CCDC22 influences integrin (ITGB1) recycling, linking endosomal sorting to cell adhesion and migration.
In HeLa cells, CCDC22 disruption impairs endosomal sorting, leading to altered copper efflux and Notch signaling. Given the HPV18-driven oncogenic background, this model enables dissection of how endosomal trafficking intersects with tumor cell biology. CCDC22 mutations are associated with X-linked intellectual disability and Ritscher-Schinzel syndrome, and its role in copper homeostasis makes this model relevant for studying copper metabolism disorders and cancer drug resistance mechanisms.
Researchers can use these polyclonal knockout cells to conduct copper uptake/efflux assays, Notch signaling reporter assays, co-immunoprecipitation of CCC complex members, and immunofluorescence with endosomal markers (EEA1, Rab5, Rab7). The heterogeneous population supports flow cytometry for surface receptor recycling, migration/invasion studies, and RNA-seq transcriptomic profiling. These applications facilitate investigation of endosomal sorting, copper homeostasis, Notch signaling, and disease modeling. For further information, please contact Ascent Research.
