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مقاله Abstract


Title: Neural differentiation of mouse retinal pigment epithelial cells by expression of optogenetic construct
Author(s): Hoda Shams Najafabadi 1, Zahra-Soheila Soheili1, Hamid ahmadieh2, Sharam Samiei3, Ehsan Ranaei Pirmardan4, Amirhossein Mahmoodi2.
Presentation Type: Poster
Subject: Retina and Retinal Cell Biology
Others:
Presenting Author:
Name: Hoda Shams Najafabadi
Affiliation :(optional) Department of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
E mail: shams.hoda250@gmail.com
Phone:
Mobile: 0910201322
Purpose:

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 differentiation in candidate cells. This study aims to induce neural differentiation of RPE cells by Opto-mGluR6 expression.

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.

Attachment: 39Neural differentiation of mouse retinal pigment epithelial cells by expression of optogenetic construct - Copy.pptx





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