The KNSTRN Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population derived from the human liver adenocarcinoma cell line SK-HEP-1, engineered to disrupt the KNSTRN gene. This product provides a heterogeneous pool of cells with targeted loss-of-function mutations in KNSTRN, enabling studies of kinetochore biology and chromosome segregation without the clonal selection artifacts associated with single-cell-derived knockouts.
The SK-HEP-1 cell line was originally established from the ascitic fluid of a patient with liver adenocarcinoma and has since been widely employed as a model for hepatocellular carcinoma and tumor microenvironment interactions. Notably, SK-HEP-1 cells exhibit a mixed phenotype with both endothelial-like and epithelial characteristics, making them particularly useful for investigating cancer cell plasticity, metastatic dissemination, and angiogenic mimicry. Their genetic background and well-characterized in vitro growth properties provide a robust platform for functional genomics and drug response studies in a liver-derived cancer context.
KNSTRN is a core component of the Astrin-SKAP kinetochore complex that stabilizes spindle microtubule attachments and silences the spindle assembly checkpoint (SAC) upon proper chromosome alignment. It interacts directly with SPAG5/Astrin and SKAP, and its activity is regulated by mitotic kinases including PLK1, Aurora B, and CDK1-Cyclin B. This regulation involves dynamic phosphorylation events that modulate the complex??s affinity for microtubules. Downstream, the complex influences SAC effectors and the APC/C to ensure timely anaphase onset and prevent aneuploidy. Additional interacting partners such as CENP-E and the NDC80 complex further integrate KNSTRN into the broader kinetochore machinery.
Disrupting KNSTRN in the SK-HEP-1 background offers a robust model for studying chromosomal instability in hepatocellular carcinoma. Loss of KNSTRN is expected to exacerbate mitotic defects, promoting aneuploidy and heterogeneity relevant to tumor evolution and drug resistance. This system is suited to explore how SAC dysfunction drives metastasis in liver cancer and, since KNSTRN mutations are found in cutaneous squamous cell carcinoma, can be used to evaluate mitotic-targeted therapies in a pan-cancer context.
These polyclonal knockout cells support diverse applications including mitotic drug sensitivity profiling via clonogenic and flow cytometry assays, live-cell imaging of chromosome segregation, and co-immunoprecipitation to map KNSTRN interaction networks. The heterogeneous polyclonal pool avoids clonal artifacts and enhances representation of diverse knockout phenotypes. Transcriptomic analysis by RNA-seq and immunofluorescence of kinetochore proteins further enable mechanistic studies and phenotypic screens. For additional details or ordering information, please contact Ascent Research.
