ABSTRACTBackground: The characteristic progression of Lewy pathology in Parkinson's disease likely involves intercellular exchange and the accumulation of misfolded α‐synuclein amplified by a prion‐like self‐templating mechanism. Silencing of the α‐synuclein gene could provide long‐lasting disease‐modifying benefits by reducing the requisite substrate for the spreading aggregation.Objectives: As a result of the poor penetration of viral vectors across the blood–brain barrier, gene therapy for central nervous system disorders requires direct injections into the affected brain regions, and invasiveness is further increased by the need for bilateral delivery to multiple brain regions. Here we test a noninvasive approach by combining low‐intensity magnetic resonance–guided focused ultrasound and intravenous microbubbles that can transiently increase the access of brain impermeant therapeutic macromolecules to targeted brain regions.Methods: Transgenic mice expressing human α‐synuclein were subjected to magnetic resonance–guided focused ultrasound targeted to 4 brain regions (hippocampus, substantia nigra, olfactory bulb, and dorsal motor nucleus) in tandem with intravenous microbubbles and an adeno‐associated virus serotype 9 vector bearing a short hairpin RNA sequence targeting the α‐synuclein gene.Results: One month following treatment, α‐synuclein immunoreactivity was decreased in targeted brain regions, whereas other neuronal markers such as synaptophysin or tyrosine hydroxylase were unchanged, and cell death and glial activation remained at basal levels.Conclusions: These results demonstrate that magnetic resonance–guided focused ultrasound can effectively, noninvasively, and simultaneously deliver viral vectors targeting α‐synuclein to multiple brain areas. Importantly, this approach may be useful to alter the progression of Lewy pathology along selected neuronal pathways, particularly as prodromal PD markers improve early diagnoses. © 2018 International Parkinson and Movement Disorder Society