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Introduction: Adenoviral gene transfer is one of the most reliable methods for introducing genes into mammalian cells. Since adenovirus infection is not cell-cycle dependent, it is possible to deliver genes of interest into primary as well as transformed cell lines. VPS35 (Vacuolar Protein Sorting 35) is a subunit of the retromer, a coat-like protein complex involved in the recycling of membrane proteins from endosomes to the trans-Golgi network.Pathological VPS35 mutants are responsible of familial forms of Parkinson's disease (PD). Aims: Production of recombinant adenovirus for VPS35 expression to study the pathological role of VPS35 in Parkinson's disease. Methods: Construction of recombinant adenovirus by using Adeno-X Expression System 1, a simple ligation-based system to incorporate a mammalian expression cassette into a replication-incompetent (llE1/llE3) human adenoviral type 5 (Ad5) genome. Results: VPS35WT or 0620N recombinant adenoviruses were successfully produced by using Adeno-X Expression System I. Transduction of VPS35 recombinant adenovirus into neuronal cells (SH-SY5Y) determines an high expression level of WT (wild type) or D620N VPS35. Conclusion: To date, many evidences have indicated the VPS35 involvement in the pathology of neurodegenerative diseases and in particular in PD. The VPS35WT or oszoN recombinant adenoviruses will be auseful tool to study the molecular mechanisms by which VPS35 induces neuronal death, and more in general will help to clarify the role of VPS35 in PD. 

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