Cell and Tissue Journal

Abstract Aim: The aims of this study were subcloning of Dj-1(PARK7) gene to lentiviral transfer vector, recombinant lentiviruses production and target cells infection with these viruses.
Material and methods: DJ1 gene obtained from pcDNA-DJ1 vector using restriction enzymes EcoR1 and Xho1. Lentiviral transfer vector was coincidental digested using EcoR1 and Sall enzymes. DJ-1 gene was interred into the lentiviral transfer and upstream of Jred gene using T4-DNA ligase. As DJ1-IRES-Jred sequence was placed in downstream and control of CMV promoter. To production of recombinant lentivruses, the made vector is coincidental transferred into the HEK-293T (Human Embryonic Kidney) cells with two packaging and envelope lentiviral vectors. Produced virus was used for target cells infection.
Results: For integrity of subcloning, enzymatic tests and PCR were used.
Fluorescent microscopy also used to show expression of Jred reporter gene that showed the success of our gene transferring .Then RT-PCR was done for showing overexpression of  DJ1 gene in transduced cells compare with normal cells.
Conclusion: This study show the lentiviral vectors success in gene transferring to eukaryotic cells and clear that this vectors can be used in treatment of nervous system diseases.