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Construction of a phage M13-based gene carrier and examination of its capacity for transgene delivery and expression in eukaryotic AGS cell line
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Mohammad Khalaj-Kondori   1, Seyedeh Elham Abedheydari2 , Mohammad Ali Hosseinpour-faizi2 |
1- University of Tabriz , khalaj@tabrizu.ac.ir
2- University of Tabriz |
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Abstract: (10258 Views) |
| Background and aim: Due to some drawbacks associated with gene delivery vehicles including viral and non-viral vectors, scientists have continued their efforts to find an ideal gene delivery vehicle. Bacteriophages have been proposed as an attractive alternative gene delivery vehicle in view of their advantages such as protection of transgene, lack of immunogenicity and stability in different conditions. This study has been conducted with the aim of obtaining a construct based on M13 phage and evaluating its capability for delivering transgene to eukaryotic cells.
Material and Method: pCMV-Script EX phagemid was constructed by intracellular excision of Lambda Zap CMV XR vector bearing GFP gene. Packaging of the construct using helper phage resulted in M13-GFP phage particles which used for transfection of human AGS cell line. Finally internalization of the phage particles into the AGS cell line was evaluated by PCR and florescence microscopy.
Results: Examination of the treated cells with florescence microscopy indicated that M13-GFP phage particles were able to internalize and express the transgene in eukaryotic cells, but the efficiency of this trasfer was very low. PCR analysis showed that internalization of the M13 GFP gene vehicle to eukaryotic cells was dose dependent.
Conclusion: The results indicated that M13 phage could be an appropriate gene delivery vehicle, because it had trivial tropism for eukaryotic cells. This means that after displaying or coupling of appropriate targeting molecules on the surface of phage particles, transgene can effectively be delivered to the target cells.
Key words: Gene therapy, Gene delivery, M13 phage.
Received: Sep 29, 2013 Accepted: Jan 20, 2014 |
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| Keywords: Gene therapy, Gene delivery, M13 phage. |
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Full-Text [PDF 257 kb]
(4618 Downloads)
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Type of Study: Original Research |
Subject:
General
Received: 2014/06/9 | Accepted: 2014/06/9 | Published: 2014/06/9
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