The CCDC97 Knockout HGC-27 Polyclonal Cells product provides a CRISPR/Cas9-engineered polyclonal knockout cell population derived from the HGC-27 human gastric adenocarcinoma cell line, featuring targeted disruption of the CCDC97 gene. This loss-of-function model enables investigation of DNA interstrand crosslink (ICL) repair and the Fanconi anemia pathway in a poorly differentiated gastric cancer background. The polyclonal knockout format, generated by CRISPR/Cas9-mediated gene disruption, yields a heterogeneous pool of knockout alleles, avoiding the artifacts of single-cell cloning while preserving the diversity of editing outcomes. This product is designed for researchers studying genome maintenance mechanisms, DNA damage responses, and the consequences of defective ICL repair in cancer cells.
The parental HGC-27 cell line was established from the ascites of a patient with metastatic gastric adenocarcinoma and is characterized as poorly differentiated and TP53-mutant. This genetic background impairs multiple DNA damage response pathways, creating a sensitized cellular context for the study of additional DNA repair defects. HGC-27 cells exhibit chromosomal instability and altered proliferation kinetics, making them a relevant model for aggressive gastric carcinoma. Combined with CCDC97 knockout, this system permits dissection of how Fanconi anemia pathway dysfunction cooperates with p53 deficiency to drive genomic instability and therapeutic vulnerabilities in gastric cancer.
CCDC97 encodes a protein that functions as an essential component of the Fanconi anemia core complex, mediating the repair of DNA interstrand crosslinks. In response to ICLs, ATR kinase activates the pathway, promoting core complex assembly. CCDC97 interacts directly with FANCI and FANCD2, facilitating their monoubiquitination and recruitment to chromatin. Downstream, this enables assembly of homologous recombination factors BRCA2 and RAD51 for error-free repair. Thus, CCDC97 links damage recognition by the core complex to FANCD2 recruitment and homology-directed repair. Disruption of CCDC97 prevents FANCD2 foci formation, causing persistent ICL lesions and heightened sensitivity to crosslinking agents including cisplatin and mitomycin C.
In the TP53-deficient HGC-27 background, ablation of CCDC97 creates a dual-hit model of genome instability that mirrors the multistep accumulation of DNA repair defects observed in advanced gastric cancer. This combination severely compromises the DNA damage response, leading to heightened chromosomal aberrations and synthetic lethality with certain chemotherapeutics. The model is particularly valuable for exploring how Fanconi anemia pathway deficiency affects tumor cell survival under replication stress, and for identifying potential therapeutic targets in p53-mutant gastric tumors with defective interstrand crosslink repair.
This knockout cell product supports immunofluorescence detection of FANCD2 foci, clonogenic survival assays to quantify crosslinking drug sensitivity, comet assays for DNA damage levels, and metaphase spread analysis for chromosomal aberrations. RNA-seq experiments and complementation with wild-type CCDC97 enable transcriptomic and functional rescue studies. These approaches underpin drug screening, pathway dissection, and translational research in gastric cancer. For further technical information, please contact Ascent Research.
