Intra-neuronal alpha-synuclein (alpha SNCA) aggregation are the leading cause of dopaminergic neuron degeneration in Parkinson's disease (PD). Most PD patients is linked with alpha SNCA gene mutations. Gene therapy shows therapeutic potential by packing gene into viral vectors to improve gene expression through stereotactic brain injections. However, through intracranial injection, the gene expression is typically limited with tissue distribution tightly adjacent to the injection track, when expressing therapeutic genes for a wider CNS region is preferable. We use microbubble-facilitated ultrasound pulsations (MB-USP) as a new gene delivering tool to enhance the limit gene delivery of local injection in brain and evaluate the feasibility using alpha SNCA as model gene. We demonstrate that MB-USP can transfect naked constructs DNA of alpha SNCA gene into two types of neuron cells and enhance the gene expression. We confirm alpha-synuclein fusion protein functionality, showing that alpha-synuclein fusion protein significantly reduce the mitochondrial activity. We show MB-USP improves in vivo gene transfer in the brain with naked construct local injection, significantly enhances alpha-synuclein expression level to 1.68-fold, and broaden its distribution to 25-fold. In vivo fused alpha-synuclein protein aggregation is also found in gene-injected mice brains by MB-USP. MB-USP provides an alternative to alpha-synuclein over expression in vitro and in vivo model for investigation of alpha-synuclein related PD therapeutic strategies. (C) 2019 Elsevier Inc. All rights reserved.