1. Department of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology 2. Department of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology 3. 2- Ophthalmic Research Center, Shahid Beheshti University of Medical Sciences 4. 3- Blood Transfusion Research Center High Institute for Research and Education in Transfusion Medicine 5. 4- Department of Molecular Genetic, Faculty of Biological Sciences, Tarbiat Modares University
Abstract
Introduction
In retinal degenerative disorders, such as age-related macular degeneration and retinitis pigmentosa neural retinal cells are damaged. neural retinal cell transplantation is one of the most promising therapeutic approaches in treatment of vision loss. retinal pigment epithelia (rpe) is a monolayer of highly specialized hexagonal-shaped cells, located between the neurosensory retina and the choroid. rpe cells represent stem cell like behaviors in cell culture. they could differentiate into neural retinal cells. optogenetic tools activate relative signaling pathway and culminate to neural differentiationin candidate cells. this study aims to induce neural differentiation of rpe cells by opto-mglur6 expression as an optogenetic tool.
Methods
Opto-mglur6 gene was cloned into paav-mcs-ires-egfp plasmid. to check the accuracy of cloning, the resultant constructs were subjected to digestion and sequencing experiments. mouse rpe (mrpe) cells were transfected with the constructs using calcium phosphate precipitation method. after 48 hours, expression of constructs was examined.
Results
Paav-mcs-ires-egfp-opto-mglur6 plasmid was constructed. sequence analysis revealed the true identity of the construct. gene expression for opto-mglur6 was confirmed in transfected mrpe cells.
Conclusion
Opto-mglur6 expression, in mrpe cell culture was successfully established.
