Isolated limb perfusion of rAAV8.CMV.eGFP in cynomologous macaques (4–5 kg) resulted in widespread gene expression in lower limb muscles. Using a custom (PE 50 Tubing) made catheter and the aid of a 0.018 m guide wire and fluoroscopy, 2 × 1012 vg/kg (2 ml volume) was administered via the femoral artery. A tourniquet occluded flow during a 10 minute dwell time to allow vector to bind. Three weeks post transfer, robust transduction was observed in muscles perfused by peroneal vessels (TA and EDL) and tibial vessels (Med and Lat. Gas. – medial and lateral gastrocnemius and soleus) (n = 2). Scale Bar, 100 μm.