The BCR Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the BCR gene in the SK-HEP-1 liver sinusoidal endothelial-like cell line. This heterogeneous loss-of-function model avoids clonal selection bias, providing a robust tool for investigating BCR-dependent pathways. The mixed population carries diverse CRISPR-mediated mutations at the BCR locus, enabling reproducible mechanistic studies in an endothelial context.
SK-HEP-1 is a human hepatocellular carcinoma-derived cell line exhibiting endothelial-like morphology and functional properties, including fenestration, endocytosis, and expression of endothelial markers. It recapitulates key features of liver sinusoidal endothelial cells, such as filtration and immune regulation. Its endothelial character and liver origin make it an ideal host for studying signaling pathways governing vascular biology, barrier integrity, and liver pathology.
BCR functions as a dual RhoGEF and RhoGAP, activating RhoA while inactivating Rac1 and Cdc42 to regulate actin cytoskeleton dynamics, cell adhesion, and migration. Upstream signals from Src kinases, integrin ??1, PDGF receptor, EGF receptor, and G??12/13-coupled receptors stimulate BCR activity. Downstream, BCR-driven RhoA activation triggers ROCK-LIMK-cofilin phosphorylation, whereas Rac1/Cdc42 inactivation attenuates PAK signaling. BCR interacts with ABL1, GRB2, NCK, 14-3-3 proteins, and F-actin, forming complexes that fine-tune cytoskeletal responses. While the BCR-ABL1 fusion drives leukemia, native BCR??s GEF/GAP activity is critical in non-hematopoietic cells.
In liver sinusoidal endothelial cells, BCR modulates permeability, endocytosis, and leukocyte transmigration by controlling Rho GTPases. Knocking out BCR in SK-HEP-1 allows dissection of these processes, particularly how BCR responds to mechanical and paracrine cues from hepatocytes or tumor cells. This model can reveal BCR??s role in sinusoidal dysfunction during liver fibrosis and its contribution to metastatic dissemination. By eliminating BCR, researchers can separate its scaffolding and enzymatic functions from ABL1 kinase signaling, advancing understanding of liver cancer and vascular pathology.
Applications include Rho GTPase activation assays (G-LISA), western blot analysis of phospho-PAK and phospho-cofilin, phalloidin staining for F-actin, transwell migration/invasion assays, and co-immunoprecipitation of BCR complexes. BCR Knockout SK-HEP-1 Polyclonal Cells enable studies of endothelial barrier function, tumor-endothelial interactions, and migration in a liver sinusoidal context. For inquiries and ordering details, please contact Ascent Research.
