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Using an unbiased anti\u2010miR lentiviral screen, miR\u201023b was identified as a critical negative regulator of BBB integrity in brain endothelial cells (BECs). Targeted inhibition of miR\u201023b using anti\u2010miR\u201023b, validated by Wnt\u2010reporter, Wnt\u2010rescue, and eCLIP analysis, demonstrates that BEC miR\u201023b silencing, enhances junctional protein expression and strengthens barrier function while suppressing transcellular transport. Global multi\u2010omics analysis reveals that miR\u201023b acts as a robust driver of angiogenesis\u2010specific genes and proteins. Conversely, anti\u2010miR\u201023b induces the expression of factors essential for BBB stabilization and repair. Notably, anti\u2010miR\u201023b enhances these protective barrier properties through a multi\u2010faceted regulation of Wnt\/\u03b2\u2010catenin, TGF\u2010\u03b2, Notch, and VEGF signaling mechanisms. Using a 3D microfluidic platform, anti\u2010miR\u201023b was shown to foster BBB repair by accelerating vessel maturation and enhancing resilience against ischemic injury. Proof\u2010of\u2010concept studies show that BEC\u2010specific AAVBR1\u2010anti\u2010miR\u201023b gene therapy reinforces BBB tight junctions, reduces BBB leakage, and improves outcome measures in a transient middle cerebral artery occlusion (t\u2010MCAO) stroke model. 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