Arak University Cell and Tissue Journal 2228-7035 4 winter 2014 03 01 Production of carrying DJ-1 gene recombinant lentiviruses and their transition to human cells Production of carrying DJ-1 gene recombinant lentiviruses and their transition to human cells 381 388 4981 10.52547/JCT.4.4.381 FA Journal Article 2014 03 16 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. <br />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. <br />Results: For integrity of subcloning, enzymatic tests and PCR were used. <br />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. <br />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. 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. <br />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. <br />Results: For integrity of subcloning, enzymatic tests and PCR were used. <br />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. <br />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. https://jct.araku.ac.ir/article_4981_8396b14c5dff55d13eea57487bf8ed26.pdf