The KIF5B Knockout SK-HEP-1 Polyclonal Cells are a genetically engineered cell pool derived from the SK-HEP-1 human hepatic adenocarcinoma cell line through CRISPR/Cas9-mediated disruption of the kinesin family member 5B (KIF5B) gene. This product comprises a polyclonal population of cells harboring targeted gene disruptions, providing a loss-of-function model for investigating KIF5B-dependent processes. As a pool of edited cells, it reflects the heterogeneous knockout effects typically observed in CRISPR-edited populations, allowing robust assessment of KIF5B function without clonal selection artifacts. The cells retain the parental line??s dual epithelial and endothelial characteristics and are suitable for a wide range of downstream analyses, including biochemical, imaging, and functional assays in biomedical research.
SK-HEP-1, originally isolated from the ascitic fluid of a patient with liver adenocarcinoma, is a widely used hepatic cancer cell line that displays both epithelial and endothelial traits. This unique phenotypic duality makes it particularly valuable for studies of tumor cell plasticity, angiogenic mimicry, and metastatic behavior in hepatocellular carcinoma (HCC). The cell line is well-characterized for its rapid proliferation, tumorigenicity in xenograft models, and susceptibility to chemotherapeutic agents. It serves as a key platform for exploring liver cancer biology, including mechanisms of invasion, drug resistance, and epithelial-to-mesenchymal transition. The SK-HEP-1 background provides a clinically relevant context for evaluating the role of KIF5B in hepatic tumor progression.
KIF5B encodes the heavy chain subunit of kinesin?1, a plus?end?directed microtubule motor protein responsible for anterograde transport of vesicles, organelles, and signaling complexes. This motor is integral to intracellular trafficking, mitotic spindle organization, cell polarity establishment, and directed cell migration. KIF5B activity is regulated by upstream kinases such as CDK1, PLK1, CaMKII, and components of the PI3K?AKT and MAPK pathways, and transcriptionally influenced by hypoxia?inducible factors and NRF2. It interacts with light chains KLC1/2 and adaptors including TRAK1/2, JIP1/3, HAP1, and GRIP1 to mediate cargo selectivity. Key downstream cargoes include mitochondria, lysosomes, APP, MAP2, Tau, VEGF mRNA, and growth factor receptors EGFR and MET, placing KIF5B at a nexus of metabolic, signaling, and neurodegenerative pathways.
In the SK-HEP-1 hepatic adenocarcinoma model, disruption of KIF5B has significant consequences for cancer cell behavior. Loss of kinesin?1 motor function impairs the directed transport of mitochondria and lysosomes, leading to altered energy metabolism and autophagic flux. It also disrupts the trafficking of EGFR and MET to the cell surface, attenuating downstream PI3K?AKT and MAPK signal transduction that normally drive proliferation, survival, and motility. Consequently, KIF5B knockout cells exhibit reduced migration and invasion capabilities, as demonstrated by wound healing and transwell assays. These defects underscore the gene??s role in metastatic dissemination and highlight the potential of this model for dissecting the molecular underpinnings of HCC progression and for identifying vulnerabilities in KIF5B-dependent tumors.
This knockout cell pool is an advanced tool for mechanistic studies of microtubule?based intracellular transport and its interplay with oncogenic signaling. Researchers can employ live?cell imaging to track organelle movement deficits, immunofluorescence to assess microtubule co?localization, and co?immunoprecipitation to map disrupted kinesin?cargo interactions. Western blotting and RT?qPCR validate KIF5B ablation, while drug sensitivity profiling and RNAi rescue experiments enable screening for targeted therapies. The cells are also invaluable for modeling KIF5B?related neurodegenerative conditions such as hereditary spastic paraplegia and amyotrophic lateral sclerosis when combined with neuronal differentiation protocols. For additional information or technical inquiries, please contact Ascent Research.
