Stable Transfection of pEGFP-N1-MOG Plasmid to Utilize in Multiple Sclerosis Gene Therapy

Authors

  • Ali Asghar Keramatinia Organization
  • Ayad Bahadori Monfared Organization
  • Hadi Azimi Organization
  • Hojjat-Allah Abbaszadeh Department of Biology and Anatomical Sciences, Shahid Beheshti University of Medical Sciences and Health Services, Tehran, Iran.
  • Maryam Sadat Khoramgah Organization
  • Mohammad Hassan Karimfar Organization
  • Mohsen Noorozian Organization
  • Navid Ahmadi Rozbahani Organization
  • Reza Mastery Farahani Organization
Abstract:

Introduction: Multiple sclerosis (MS) is a disease of the immune system: it attacks the myelin around the axons and leaves them exposed. Destruction of myelin weakens the electrical conduction of ions and thus leads to a lack of communication in the nervous system.  Methods: In the present study, we constructed recombinant plasmid and then transformed to E. coli cell. The colonies containing plasmid were selected by Colony PCR. Enzyme digestion and sequencing were utilized to approve the accuracy of the extracted plasmid of these clones. Recombinant plasmid transfect in to mesanchymal stem cells. Results: Plasmid was verified correctly. After transfection, the transcription of MOG gene and the expression of MOG protein were proved by RT-PCR, western blotting and Elisa. Conclusion: Plasmid was constructed correctly and mesenchyme stem cells were successfully transfected by transfection and protein can be expressed well, setting a proper foundation for the future studies on the transplantation of gene modified mesanchymal stem cells in order to promote Multiple sclerosis.

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Journal title

volume 12  issue 1

pages  3- 8

publication date 2015-02

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