The ITGB2 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 host cell line. This heterogeneous pool features targeted disruption of the ITGB2 gene, achieved through CRISPR/Cas9-mediated mutagenesis, yielding a loss-of-function model for beta-2 integrin subunit deficiency. The polyclonal format preserves genetic diversity and is particularly advantageous for large-scale functional genomic screens and assays where uniform knockout across a population is sufficient.
The SK-HEP-1 cell line, originally established from the ascites fluid of a patient with liver adenocarcinoma, displays hallmark endothelial characteristics, including cobblestone morphology, expression of endothelial markers, and the ability to support leukocyte adhesion and transmigration. These properties have established SK-HEP-1 as a robust in vitro model of liver sinusoidal endothelium, widely employed in research on hepatic angiogenesis and the extravasation of metastatic tumor cells.
ITGB2 encodes the integrin beta-2 subunit (CD18), which obligatorily heterodimerizes with alpha-L (ITGAL), alpha-M (ITGAM), alpha-X (ITGAX), or alpha-D (ITGAD) to form leukocyte adhesion receptors. These integrins engage intercellular adhesion molecules ICAM1 and ICAM2 on endothelial surfaces, with intracellular activation modulated by Talin and Kindlin-3. Ligand binding triggers outside-in signaling cascades involving phosphorylation of SRC family kinases and focal adhesion kinase (FAK), leading to activation of downstream effectors including Paxillin (PXN), RAC1, and CDC42, which drive actin polymerization and cell migration. The pathway is responsive to upstream inflammatory stimuli such as TNF-alpha, IL-1beta, and CXCL12, which upregulate integrin expression via NF-kB-mediated transcription.
In the SK-HEP-1 background, ITGB2 knockout disrupts the fulcrum of leukocyte-endothelial interaction, providing a powerful platform to dissect the pathophysiology of leukocyte adhesion deficiency type 1 (LAD-1). The model recapitulates deficits in firm adhesion and transmigration that are central to systemic lupus erythematosus, rheumatoid arthritis, and inflammatory bowel disease. Moreover, within the liver sinusoid context, this knockout enables mechanistic studies of how loss of beta-2 integrin-dependent ICAM-1 engagement on endothelial cells alters the behavior of circulating immune and cancer cells, shedding light on tumor-immune crosstalk and metastatic niche formation.
This polyclonal knockout resource supports a wide range of assays, including static and flow-based adhesion assays to measure leukocyte attachment, transmigration assays across endothelial monolayers, and flow cytometry to verify loss of integrin surface expression. Downstream signaling perturbations can be analyzed via immunofluorescence and Western blotting, targeting molecules such as FAK, PXN, and phospho-SRC. The model is ideal for phenotypic compound screens aimed at identifying modulators of leukocyte adhesion or agents that bypass integrin deficiency, and for unbiased secretome profiling of liver endothelial cells. For comprehensive product information, please contact Ascent Research.
