The EFCAB14 Knockout Huh-7 Polyclonal Cells are a heterogenous population of Huh-7 hepatocellular carcinoma cells subjected to CRISPR/Cas9-mediated disruption of the EFCAB14 gene. This polyclonal knockout pool offers a genetically diverse loss-of-function model for studying EFCAB14-dependent processes in a liver cancer context. The editing targets the EF-hand calcium-binding domain, impairing functional protein production, as confirmed by genomic and protein analyses. The polyclonal format avoids clonal bias, providing a robust tool for functional genomics inquiry into calcium signaling modulators.
Huh-7 is a well-differentiated human hepatocellular carcinoma line retaining key hepatic epithelial features, including metabolic pathways and hepatitis C virus permissiveness. As a widely used model in liver cancer, antiviral research, and drug metabolism studies, Huh-7 cells proliferate rapidly and are amenable to gene-editing and transfection. They express a full complement of calcium channels, pumps, and regulatory proteins typical of liver epithelia, offering a physiologically relevant platform for studying calcium signaling perturbations in hepatocyte-derived carcinoma cells.
EFCAB14 encodes an EF-hand calcium-binding protein that senses intracellular calcium fluctuations. Its activity is modulated by upstream factors such as cytosolic calcium, calmodulin, and GPCR signaling. Upon calcium binding, EFCAB14 interacts with calmodulin, S100 proteins, and calcium channel subunits, forming complexes that regulate downstream targets including calcium-dependent enzymes, CAM kinases (e.g., CAMKII), and calcineurin. Through these interactions, EFCAB14 participates in IP3 receptor-mediated calcium release and subsequent NFAT activation, thereby influencing calcium-dependent gene expression and cellular responses. Knockout of EFCAB14 interrupts this signaling network, disrupting calcium homeostasis.
In hepatocellular carcinoma, EFCAB14 knockout likely perturbs calcium-dependent processes governing tumor cell behavior. Loss of EFCAB14 impairs calcium signaling dynamics, potentially affecting proliferation, apoptosis, and migration??key aspects of liver cancer progression. Huh-7 cells rely on precise calcium signals for cell cycle and survival; thus, EFCAB14 deficiency enables dissection of EF-hand protein contributions to malignant phenotypes. This knockout model provides a system to investigate how calcium-binding protein disruption reprograms signaling networks linked to hepatocellular carcinoma and calcium dysregulation disorders, identifying potential vulnerabilities.
These polyclonal knockout cells support diverse applications, including calcium signaling studies in liver cancer, functional screening of EF-hand family proteins, and tumor microenvironment calcium dynamics. Typical assays encompass western blotting and RT?qPCR for knockout validation, immunofluorescence for protein localization, calcium imaging for real-time flux monitoring, and phenotypic assays for migration, invasion, apoptosis, and proliferation. The EFCAB14 knockout Huh-7 polyclonal population is a versatile loss-of-function resource for investigating EF-hand proteins in hepatic malignancies. For further details, contact Ascent Research.
