The INF2 Knockout SK-HEP-1 Polyclonal Cells constitute a CRISPR/Cas9-mediated gene-edited human hepatic endothelial model in which the INF2 gene has been disrupted to ablate its function. Supplied as a polyclonal cell population rather than a clonal line, this product enables robust loss-of-function analyses while minimizing the effects of clonal selection artifacts. INF2 encodes a formin protein essential for actin nucleation and mitochondrial fission, positioning this knockout tool for in-depth studies of cytoskeletal and organelle dynamics.
The host SK-HEP-1 cell line was originally isolated from the ascites of a liver adenocarcinoma patient but exhibits a stable endothelial phenotype closely resembling hepatic sinusoidal endothelial cells. These cells are actively involved in endothelial barrier maintenance, angiogenic responses, and inflammatory signaling, making them a well-characterized in vitro model for hepatic microvascular biology. Their endothelial nature is validated by the expression of characteristic markers and functional behaviors including tube formation and barrier establishment.
At the molecular level, INF2 operates under the control of upstream activators RhoA, Cdc42, and calcium/calmodulin, and is recruited to ER?Cmitochondria contact sites where it cooperates with Spire1C and IQGAP1 to assemble actin filaments. This localized actin polymerization is critical for recruiting the dynamin-related GTPase Drp1, which drives mitochondrial fission. In the cytoplasmic compartment, INF2 modulates actin dynamics to regulate the activity of the MRTF-A/SRF transcriptional machinery, linking cytoskeletal remodeling to gene expression. Profilin and calmodulin further tune INF2 activity through direct interactions.
In the context of hepatic sinusoidal endothelium, INF2-mediated actin dynamics are likely central to barrier integrity, cell migration, and angiogenic sprouting. Concurrently, INF2-dependent mitochondrial fission governs mitochondrial distribution and metabolic adaptation, processes crucial for endothelial function during physiological and pathological challenges. Disruption of INF2 in SK-HEP-1 cells therefore provides a tractable system to unravel how formin-dependent cytoskeletal events intersect with mitochondrial homeostasis in the liver microvasculature.
These knockout cells are amenable to a wide array of experimental approaches, including TEER assays to quantify barrier function, migration and invasion assays to probe angiogenic behavior, and immunofluorescence imaging to assess actin and mitochondrial morphology. Biochemical assays such as western blotting for Drp1 phosphorylation, RT-qPCR for MRTF-A/SRF target genes, and co-immunoprecipitation of INF2 complexes can be performed. Disease modeling for focal segmental glomerulosclerosis and Charcot-Marie-Tooth disease with glomerulopathy, as well as screening for INF2 inhibitors, represent high-value applications. For further technical inquiries, please reach out to Ascent Research.
