Overexpression of α-Synuclein inSHSY5Y cell to generate a model for Parkinson’s disease

The aggregation of α-synuclein (α-Syn), a protein found at high concentration in the brain, has been shown to be involved in Parkinson’s disease (PD) and other α-synucleinopathies. To unravel the complex pathological processhappening in PD, developing the controllable cellular models is welcome worldwide including numerous α-Syn based models. One of the events that cause PD is the overexpression of α-Syn and so making a cell model using the high expression of the heterologous protein could be useful in the studies related to the mechanism of PD as well as related pharmaceutical investigation. SH-SY5Y cells possess a complete dopaminergic system that uses widely for modeling PD. Herein, we used a lentiviral vector to overexpress α-Syn in SH-SY5Y cells as a model of PD due to it s high-level and long-term transgene expression. Accordingly, αSN cDNA complemented with Kozak sequence was cloned into the pLEX-JRed- TurboGFP lentiviral plasmid (Stem Cell Technology Research Center). The obtained construct along with packaging and envelope plasmids were transfected to the HEK293T cell line. The supernatant of the cell culture medium was collected and concentrated using PEG6000. HEK293T cells were infected with concentrated viruses and the viral titer was determined. SHSY5Y cells overexpressing α-Syn were prepared using infection with obtained lentiviruses. This cell line can be used for more pharmaceutical investigation and more genetic mechanism of PD.

کلیدواژه ها:

Alpha-synuclein ، Parkinson disease ، SHSY5Y cell ، HEK293T