The Cdh5 Knockout bEnd.3 Cell Line is a CRISPR/Cas9-engineered mouse endothelial cell model in which the Cdh5 gene has been disrupted to eliminate functional CDH5/VE-cadherin expression. This gene-edited line is generated in bEnd.3 cells, an immortalized murine brain microvascular endothelial cell line, and provides a stable in vitro system for studying the consequences of endothelial adherens junction loss in a cerebral microvascular context. As VE-cadherin is a central structural and signaling component of endothelial cell-cell contacts, this model is particularly relevant for mechanistic studies of barrier regulation and vascular inflammatory responses.
bEnd.3 cells are widely used as an experimental model of brain-derived microvascular endothelium and blood-brain barrier-associated endothelial biology. They recapitulate key aspects of cerebral endothelial behavior, including monolayer formation, permeability control, and responses to inflammatory and angiogenic stimuli. Because brain microvascular endothelial cells are essential for vascular barrier formation and regulate leukocyte trafficking, permeability, and angiogenic remodeling, bEnd.3 provides a useful background for investigating neurovascular dysfunction, including processes relevant to stroke, neuroinflammation, cerebral edema, and blood-brain barrier disruption.
CDH5 encodes VE-cadherin, an endothelial-specific transmembrane cadherin that mediates homophilic adhesion at adherens junctions and links adjacent endothelial cells to catenin-associated cytoskeletal networks. VE-cadherin forms complexes with CTNNB1, JUP, and CTNND1 and functionally couples junctional architecture to ACTB-associated cortical actin organization through factors including ACTN1 and VCL. Its junctional stability is regulated by upstream inputs such as VEGF-A/VEGFR2 (KDR), TNF, IL1B, thrombin, TGF-??, shear stress, and SRC family kinases including FYN and YES1. VE-cadherin also interacts with PECAM1, TJP1, PTPRB/VE-PTP, and signaling nodes such as SRC, RAC1, and RHOA that coordinate endothelial barrier integrity, paracellular permeability, and angiogenic sprouting. Loss of CDH5 is therefore expected to perturb adherens junction organization, alter ??-catenin-associated signaling, and modify leukocyte transendothelial migration and permeability-related phenotypes.
In the bEnd.3 background, Cdh5 disruption provides a biologically relevant system for examining how endothelial junction failure alters brain endothelial behavior. This context is valuable for defining pathway dependencies that connect junctional adhesion to VEGF signaling, cytoskeletal remodeling, inflammatory stimulation, and monolayer integrity. The model may support studies of disease-associated mechanisms linked to vascular leak, tumor angiogenesis, diabetic retinopathy, sepsis-associated endothelial dysfunction, and vascular malformations, particularly where altered endothelial cohesion contributes to pathology.
This knockout cell line can be applied in western blotting, RT-qPCR, and RNA-seq workflows to evaluate gene and protein-level consequences of Cdh5 loss, as well as in immunofluorescence and flow cytometry assays to examine junctional architecture and surface marker changes. Functional applications include transendothelial electrical resistance measurements, permeability tracer flux assays, and barrier integrity assays to quantify monolayer disruption; co-immunoprecipitation and phospho-signaling analysis to assess effects on CTNNB1-, SRC-, or VEGFR2-associated complexes and signaling states; and migration, leukocyte transmigration, and tube formation assays to interrogate angiogenic and inflammatory endothelial phenotypes. Researchers may contact Ascent Research for additional technical information, product details, or related gene-edited cell models.
