Gene Expression under F8 Promoter Driving In Mouse Hepatoma Cells: A Step towards Gene Therapy of Hemophilia
Publish place: IRANIAN JOURNAL of PATHOLOGY، Vol: 6، Issue: 4
Publish Year: 1390
نوع سند: مقاله ژورنالی
زبان: English
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JR_IJP-6-4_001
تاریخ نمایه سازی: 5 آبان 1393
Abstract:
Background and Objectives: Significant progress has been made in treatment of hemophilia. Exvivo gene therapy is going popular due to the capability of this method in using isogenic cells forgenetic manipulation and reintroducing them into same host after proliferation. Most gene therapytechniques use viral vectors, which usually harbor a strong and non-specific promoter (e.g. CMV early promoter) for driving the downstream gene. This may be a disadvantage due to uncontrollable nature of gene expression. In addition, considering the potentials of recently introduced stem cellsas reservoirs and their potential to differentiate to other cell lines, uncontrolled expression may have unknown outcomes. To make gene therapy of hemophilia more resembling to the nature, wesupposed f8 promoter might be a good candidate for driving downstream f8 coding sequence.Materials and Methods: To test our hypothesis, we designed and constructed a DNA construct by PCR, which harbors EGFP coding sequence downstream to mouse f8 promoter and transfected itto a mouse hepatoma cell line. Transfection was assayed qualitatively by fluorescent microscopy.Results: Fluorescence was detected in transfected cells a sign of presence of EGFP. Conclusion: f8 promoter can drive expression of downstream genes, a capability which and mayhave potential to be used in gene therapy of hemophilia. A conclusion that should be examined by further studies.
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Authors
Pejman Fard-Esfahani
Dept. of Biochemistry, Pasteur Institute of Iran, Tehran, Iran
Mehdi Kadivar
Dept. of Biochemistry, Pasteur Institute of Iran, Tehran, Iran
Mojgan Allahyari
Dept. of Education, Pasteur Institute of Iran, Tehran, Iran
Fatemeh Mirkhani
Dept. of Education, Pasteur Institute of Iran, Tehran, Iran