Treatment of Focal Brain Ischemia with Viral Vector-Mediated Gene Transfer

Promoting functional recovery after ischemic brain injury has emerged as a potential approach for the treatment of ischemic stroke. An ideal restorative approach to enhance long-term functional recovery is to promote postischemic angiogenesis and neurogenesis. This chapter describes a system using adeno-associated viral (AAV) vector-mediated vascular endothelial growth factor (VEGF) gene transfer into the ischemic brain. The methods described here for construction, production, and purification of AAV vector expressing VEGF gene can also be applied to producing AAV vectors expressing other genes. This chapter also illustrates the methods to produce mouse middle cerebral artery occlusion (MCAO), injection of viral vector into the mouse brain, and standard assays for determining the success of brain ischemia and gene transfer.