The CCDC6 Knockout Huh-7 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the CCDC6 gene has been disrupted via CRISPR/Cas9-mediated gene editing. This product provides a heterogeneous population of Huh-7 cells carrying targeted disruption of CCDC6, enabling loss-of-function studies without clonal selection. As a polyclonal pool, it captures the diversity of editing outcomes across the population, suitable for experiments where bulk functional effects are assessed. These cells offer a versatile platform to interrogate the role of CCDC6 in DNA damage responses, apoptosis, and tumor suppression within a hepatic carcinoma background.
The host cell line, Huh-7, was originally established from a well-differentiated hepatocellular carcinoma of a 57-year-old Japanese male. This adherent epithelial cell line is widely employed in liver cancer research due to its retention of hepatocyte-like features and its permissiveness to hepatitis C virus (HCV) replication. Huh-7 cells retain wild-type p53 and express relevant signaling molecules, making them a representative model for studying hepatocarcinogenesis, drug metabolism, and viral oncology. The integration of a CCDC6 knockout into this well-characterized line creates a powerful system for dissecting pathway interactions in a disease-relevant context.
CCDC6 is a key component of the DNA damage response network, functioning as a substrate of the ATM kinase. Upon genotoxic stress, such as ionizing radiation or chemotherapeutic agents, ATM phosphorylates CCDC6, which then facilitates CREB1-mediated transcriptional programs that promote apoptosis. CCDC6 forms complexes with ATM and interacts with the phosphatase PP4C and the transcription factor CREB1, linking upstream damage signals to downstream apoptotic effectors. CCDC6 is a recognized tumor suppressor, originally identified in the RET/PTC1 fusion oncogene, and its inactivation is implicated in various cancers. This protein operates at the intersection of genome maintenance and cell death.
In Huh-7 hepatocellular carcinoma cells, the loss of CCDC6 disrupts this DNA damage?Capoptosis axis, potentially enhancing cell survival and genomic instability under stress. This model is especially relevant given Huh-7??s p53 status and its use in HCV studies, where viral proteins may interfere with host damage responses. The CCDC6 knockout Huh-7 polyclonal cells thus provide a defined genetic background to explore how CCDC6 deficiency contributes to hepatocellular carcinogenesis, chemo resistance, and viral oncogenesis. They enable researchers to dissect CCDC6-dependent signaling without interference from wild-type protein expression.
Research applications for these cells are broad and include mechanistic studies of the ATM-CCDC6-CREB1 pathway, apoptosis regulation, and tumor suppression. Typical experimental approaches involve Western blotting for CCDC6 and phospho-ATM, RT-qPCR for CREB1 target genes, flow cytometric apoptosis assays using Annexin V/propidium iodide, comet assays to assess DNA damage, clonogenic survival assays following genotoxic treatment, and immunofluorescence detection of ??H2AX foci as a marker of double-strand breaks. Co-immunoprecipitation can be used to examine ATM-CCDC6 interactions. They are also suitable for drug sensitivity screens, synthetic lethality experiments, and functional genomics. For further information, please contact Ascent Research.
