The CCDC22 Knockout Huh-7 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout population for functional studies of CCDC22 in a human liver cancer background. This polyclonal cell population was generated by CRISPR/Cas9-mediated gene disruption, introducing loss-of-function mutations across the CCDC22 locus, resulting in a heterogeneous knockout pool suitable for pooled phenotypic assays. The knockout model enables investigation of CCDC22-dependent processes without the need for clonal isolation, maintaining representation of diverse editing outcomes.
The host cell line Huh-7 is a well-differentiated hepatocellular carcinoma line derived from a human liver tumor. It exhibits adherent, epithelial morphology and is widely employed as a model for hepatocarcinoma, hepatitis C virus replication studies, and drug metabolism research. Huh-7 cells retain many characteristics of primary hepatocytes, making them a valuable platform for studying liver-specific pathways, including metabolic regulation, detoxification, and oncogenic signaling networks.
CCDC22 encodes a protein containing coiled-coil domains that serves as a core component of the CCC (COMMD/CCDC22/CCDC93) complex. This complex cooperates with the retromer (VPS26/VPS35) and WASH complexes to mediate endosomal sorting and recycling of transmembrane receptors. Notably, CCDC22 is essential for the retrieval of copper transporters ATP7A and ATP7B, thereby maintaining copper homeostasis. Additionally, CCDC22 regulates Notch signaling by controlling recycling of NOTCH1 and affects integrin trafficking and ciliary signaling components. Molecular interactions have been demonstrated with COMMD family members (COMMD1-10), CCDC93, VPS26, VPS35, and the WASH complex.
In the context of hepatocellular carcinoma, CCDC22 knockout in Huh-7 cells offers a unique system to dissect the intersection of endosomal trafficking, copper metabolism, and developmental signaling pathways. Dysregulation of Notch signaling and copper balance has been implicated in liver cancer progression and chemoresistance. Moreover, CCDC22 mutations are associated with X-linked intellectual disability and Ritscher-Schinzel syndrome, underscoring its developmental significance. This model may help elucidate how CCC complex dysfunction contributes to hepatic pathophysiology and crosstalk between oncogenic and metabolic networks.
Researchers can utilize these cells to investigate endosomal trafficking mechanisms and receptor recycling dynamics using assays such as flow cytometry for integrin ??1 surface expression, immunofluorescence for endosomal marker colocalization, and co-immunoprecipitation of CCC complex components. Functional studies of copper homeostasis can be performed via copper accumulation assays and viability tests under copper stress. Notch signaling activity can be measured with reporter assays following ligand stimulation. The model is also suitable for drug screening to identify modulators of CCC complex function. For further technical information, please contact Ascent Research.
