The CD59 Knockout Huh-7 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population of Huh-7 hepatocellular carcinoma cells harboring targeted disruption of the CD59 gene, generating a heterogeneous knockout model for loss-of-function studies. Unlike clonal isolates, this polyclonal pool reflects the spectrum of editing outcomes, offering a robust platform to analyze CD59-dependent phenotypes without the bias of a single genetic clone. The engineered population enables investigation of complement regulatory mechanisms and immune evasion strategies in a physiologically relevant human liver cell background.
The Huh-7 cell line is a well-differentiated hepatocyte-derived carcinoma originally isolated from a liver tumor of a 57-year-old Japanese male. This epithelial cell line is widely employed as a model for hepatocyte biology, hepatitis C virus replication, and hepatocellular carcinoma pathogenesis. Huh-7 cells retain many functional characteristics of primary hepatocytes, including expression of complement regulatory proteins, making them a suitable host for studying CD59??s protective role against autologous complement attack.
CD59 is a glycosylphosphatidylinositol (GPI)-anchored glycoprotein that inhibits complement-mediated lysis by binding to C8 and C9 within the C5b-9 membrane attack complex (MAC), thereby blocking C9 polymerization and pore formation. In addition, CD59 functions as a CD2 ligand, contributing to T cell co-stimulation. Its expression is induced by inflammatory cytokines such as TNF-?? and IL-1?? through STAT3 and NF-??B signaling, and it localizes to lipid rafts where it interacts with complement components and CD2 to regulate immune effector functions.
In hepatocellular carcinoma, CD59 upregulation is a recognized mechanism of immune evasion, protecting tumor cells from complement-dependent cytotoxicity. The CD59 Knockout Huh-7 Polyclonal Cells thus provide a relevant model to dissect how loss of CD59 affects susceptibility to complement attack in a liver cancer context. This system enables the study of liver tumor cell vulnerability to complement-mediated lysis and the interplay between complement regulators and oncogenic signaling pathways in hepatocarcinogenesis.
These polyclonal knockout cells are suited for a range of experimental applications, including complement-dependent cytotoxicity assays, flow cytometric analysis of CD59 surface loss, and immunofluorescence detection of MAC deposition. Researchers can employ this model to evaluate therapeutic agents that target the complement cascade, to model aspects of paroxysmal nocturnal hemoglobinuria when combined with other GPI-anchor deficiencies, and to investigate the role of complement in hepatic immune surveillance. Co-immunoprecipitation and western blotting enable analysis of CD59??s molecular interactions with C8, C9, and CD2. For comprehensive technical support and product inquiries, please contact Ascent Research.
